CA2174933A1 - Detection of h. pylori in the stomach - Google Patents
Detection of h. pylori in the stomachInfo
- Publication number
- CA2174933A1 CA2174933A1 CA002174933A CA2174933A CA2174933A1 CA 2174933 A1 CA2174933 A1 CA 2174933A1 CA 002174933 A CA002174933 A CA 002174933A CA 2174933 A CA2174933 A CA 2174933A CA 2174933 A1 CA2174933 A1 CA 2174933A1
- Authority
- CA
- Canada
- Prior art keywords
- reagent
- urea
- carrier
- dense
- stomach
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/58—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving urea or urease
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/006—Biological staining of tissues in vivo, e.g. methylene blue or toluidine blue O administered in the buccal area to detect epithelial cancer cells, dyes used for delineating tissues during surgery
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
- A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
- A61K51/04—Organic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
- A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
- A61K51/04—Organic compounds
- A61K51/0404—Lipids, e.g. triglycerides; Polycationic carriers
- A61K51/0406—Amines, polyamines, e.g. spermine, spermidine, amino acids, (bis)guanidines
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/205—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Campylobacter (G)
Abstract
A method for the in vivo detection of urease-producing helicobacter in the upper stomach is disclosed. The dense carrier is divided into two separate groups which are combined with separate reagent indicators, one of which also contains urea. The carriers are food soluble products, preferably sugar beads having a diameter of approximately 0.2 to 3.0 mm. The treated carriers and urea are encapsulated in a soluble capsule which is administered to a patient. The density of the carriers cause the capsule to migrate to the gastric mucosa, where the capsule is dissolved, placing the reagents and urea in direct contact with the gastric mucosa. The urea reacts with any urease present in the stomach by creating ammonia, which increases the pH within the stomach. The two reagents react differently, through color change, to the increase in pH, which is viewed through use of art endoscope. A preferred first reagent is bromothymol blue (dibromothymolsulfonphthalein), which changes yellow in the presence of urease, and a preferred second reagent is phenol red (phenolsulfonphthalein) which turns red in the presence of urease.
Description
WO 95111672 ~ 9 3 3 PCI~/~S9.~112332 .
DETECTION OF H. PYL0~I IN THE ST0MACH
BACKGROUND OF THE INVENTION
Brief DescriDtion of the InY~ n~inn The instant invention relates to a novel method of in ivo diagnosis of upper gastromtes-tinal diseases.
Brief DescriDtiQn of the Prior Att Factors adversely affecting the function of the ~ ' ' system in humans are ex-ceedingly varied in their nature, Such disorders may arise in the upper or lower ~ , tracts, or both. There is a broad range of causes of ~ ' disorders, including genetic, physiologicdl, C...;l~ ' and psychogenic hctors. Accordingly, the diagnosis amd manage-ment of these disorders can be ~ difficult.
Among the chronic disorders of the upper t~ ' ' tract are those which fall under the general categories of gastritis and peptic ulcer disease. The upper Er..l~u;.lt~liuldl tract is generally defined as including the esophagus, the stomach, the duodenum, the jejunum and ileum.
Peptic ulcers are lesions of the t~ u;..~c~li..dl tract lining, ~I..,~.lc.i~d by loss of tissue due to the action of digestive acids and pepsin. It has generally been held that peptic ulcers are caused by gastric l~ C~CL;UU~ decreased resistance of the gastric lining to digestive acids and pepsin, or bo~h. Gastritis is, by definition, an ;.31~ ;,." of the stomach mucosa. In practice, though, the disorder is manifested by a broad range of poorly-defined, and heretofore inadequately treated, symptoms such as indigestion, "heart burnn, dyspepsia, and excessive eruaation.
As with the management of any disorder, the rapid, precise, and accurate diagnosis of t,~,.Lr~ ~' disorders is of paramount importance. The typical means used to diagnose the gastrointestinal disorder presented by a given patient v~ill depend upon such factors as the nature and severity of symptoms, the overall health of the individual, the medical history of the patient, the need for a specific diagnosis in order to implement a treatment with reasonable likelihood of success, and the availability of diagnostic devices. However, the diagnostic methods typically employed in the art are often slow, ~ Pr~r~ , costly, and may yield equivocal or inaccurate results. Thus, for patients not ha~ing severe symptoms, a precise diagnosis of a g~l~L-.
-.
~1 7 4933 2 disorder might not be attempted. Sueh patients may simply be treated with eonYentional therapies, sueh as with antaeids or drugs whieh inhibit stomach acid secretion. While such therapies might provide temporary symptomatic relief, a cure is ohen not effected. More effec-tive treatments may depend upon better diagnoses Df the actual underlying ~;callu;~ hlcl disor-der. In particular, it has been discovered that many sueh ~;.laLI- ' ' ' disorders are mediated by infedion of gastrie mucDsa by Helicobaeter pylori. H. pylori is a ( ;rr~ ;cl;._ spiral or-ganism whieh produces the enzyme urease. The organism is ~ / found beneath the mucus layer of the luminal aspect of the gastric epithelium and in the gastrie pits. Helicobader can be diagnosed by blood test for antibodies, breath test, or biopsy of the stomach lining. An-tibodies, however, can remain positive for many months after the baeteria have been eradicated.
The presenee of antibodies presents a falsely positive result in c~ , 10 to 15% of patients. Biopsies are relatively quick; however, they add time, expense and risk. Although rela-ti~ely minor, there is a 1 in 20,000 risk of bleeding from a biopsy site. Biopsies cannot be per-formed on patients who have a tendency to bleed, such as patients with hemophilia and liver dis-ease. Additionally, it has recently been found that helicobacter is patchy, thereby requiring mul-tiple biopsies to obtain 10û% aeeuraey. The eost for a biopsy is C~ ' $100. Biopsies also inerease the risk of the person handling the tissue being exposed to HIV. If a urease test is used, the biopsy sample must be plaeed in the test by the nurse, thereby requiring an additional person during the test.
The prior art has diselosed testing for hcall, ' disorders, the majority of which have been in vitro. Many lests have also been disclosed using urea and indicators.
Marshall, 4,748,113 diseloses eompositions and methods for the diagnosis of ~,calru~ ~lhlcl disorders in-ol-ing urease. Methods inelude obtaining a gastric sample material and eontacting the material with a composition including urease and an indicator.
Marshall 4,830,û10, discloses methods for the diagnosis of ~ lu;~ lcl disorders. The mcthod steps include - ' c~iull of urea-containing compositions prior lo assay.
2 1 7f~933 Steward et al, 5,139,934 disclose substrate ~ and method of urease assay. The method is an in vitro y that includes the use of pH indicators.
Nagatsu et al, 4,147,692 disclose methods and ~ for mcasuring enzymatic ac-tivities and correlating such activities with various disease states, Kraffczyk et al, 3,873,369 disclose colorimetric iDdicators for the .' of urea.
Vasquez et al, 4,851,209 disclose in viYo diagnostic procedures for the clinical evaluation of ' ulcer disease using radioactive isotopes. Procedures involYe prior ' ' ' ~iUI-of a diagnostic ~ l followed by scimigraphic imaging of the ~ area of interest with scintigraphic imaging equipment.
Although the use of urease or other indicators has been used in combination with pH in-dicators, all except Vasquez et al are conducted in vitro.
The instant invention discloses a method of detecting the alkaline pH change in vivo. The test dramatically cnts down the number of biopsies required and is safe for patients having any bleeding tendencies while being rapid and low cost. Addilionally, through the color change, it can be determined if the change is a true positive or a false positive reaction.
BRIEF DESCRIPTION OF THE DRAWINGS
The advantagcs of the instant disclosure will become more apparent when read with the specification and the drâwings, wherein:
FICURE 1 illustra~es the locâtion of the beads in the stomach and the urea/ammonia trans-fer.
SUMMARY OF THE INVENTION
A method for the in vivo detection of urease-producing helicobâcter in the upper stomach is disclosed. A dense carrier is used which is divided into two separate groups, the first combined with a first reagent indicator and the second combined wilh a second reagent indicator and urea, i.e. 14C-Urea or 13C-Urea. The carriers are food soluble products, preferably sugar beads having a diameter of ~ 0.2 to 3.0 mm. The carrier and reagent can be combiDed through coat-ing the carrier or combining the carrier and reagent. The treated carriers and urea are encapsu-.
21 74q33 lated in a soluble capsule which is administered to a patient. A buffer can be added, if desired, toobtain specific results. The density of the carriers cause the capsule to migrate to the g astric mucosa. The gastric juices dissolve the capsule containing the reagents and urea thereby placing the two reagents and urea combination in direct contact with the gastric mucosa. The urea reacts with any urease present in the stomach by creating ammonia, which causes the pH within the stomach to increase. The two reagents react differently, through color change, to the mcrease in pH, which is viewed through use of an endoscope. A preferred first reagent is bromothymol blue (dibrom~h~ . ' ' ' ), which changes yellow in the presence of urease, and a preferred second reagent is phenol red (I ' ' '' , ' ' ' ), which turns red in the presence of urease.
DETAILED DESC~lPrlON OF THE INVENTION
The instant disclosure uses indicators to detect alkaline pH change in the stomach during endoscopy. A change in the colors of the indicators detects pH change within the stomach. A cer-tain combination of colors will indicate the presence of helicobacter, or H. pylori, organisms.
Urea has the formula H2NCONH2 and is a naturally occurring product of protein metabolism. Gastric materials from humans or other animals having t,~ dl disorders contain relatively large quantities of urease (urea . ' ~J~ld~
which hydrolizes urea to ammonium carbonate or ammonia and carbon dioxide. Normally urease is present in the body in only trace amounts, performing the function of ' , ~ urea. H.
pylori increases the amount of urease above normal in the affected areas. The increased urease reacts with the administered urea by creating ammonia, which causes an indicator color change due to the increased alkalinity.
The indicators useful in this invention are weak acids with sharply different colors in their dissociated (ionized) and un-dissociated (neutral) states. The indicators useful herein have PKa values of from about 6.5 to about 8.5, preferably from about 7.0 to about 8Ø The color exhibited by the indicator in the present composition will depend upon the pH of the composition, the particular indicator used, and the dissociation constant (Ka) for that indicator (i.e., PKa ¦ log10Ka). As the color exhibited by the indicator changes over a ~ 74q33 range of pH values (pH=logl0 [H+]), the indicators useful in Ihe present, . chamge color over a pH range of from about 5.5 to about 9.0, preferably from about 6.5 to about 85. The pH of the present composition are accordingly adjusted to a pH at least about one pH unit lower than the PKa of the indicator used (i.e. having a hydrogen ion [H+] ten times less than (10~o of) the hydrogen ion ~ ll in a solution having a pH equal to the pKa of the imdicator).
Preferably, the pH is adjusted to a pH about two pH units below the PKa of the indicato~. Adjust-ment of the pH of the present . can be ef&cted by addition of a base (e.g. sodium hydroxide) or an acid (e.g. hydrochloric acid or citric acid). Thus, preferably, the pH of the oom-position of this invention is adjusted to a pH of from about 5.0 to about 65, with the preferred embodiment being from about 5.0 to about 6Ø
The preferred reagents are 1,.. ' ~ -' blue (d;l,.. ', ' '' . ' ' ' ' ) indicator, Reagent 1, and phenol red (I ' l '' . ' ' ' ) indicator, Reagcnt 2. Other indicators nseful herein include p-nitrophenol, neutral red (2-methyl-3-amino-6-:' ' ,' . ' ), quinoline blue (cyanine), cresol red (o-cr~ ~ , ' ' ' ), and thymol blue (lh, ~ ' ). In-dicators among those useful herein are described in the The Merk Index (9th ed. 1976), incor-porated by reference herein. Reagent 2 has urea added to react to the urease enzyme, if present.
The urea penetrates the mucus layer of the stomach to come into contact with the nrease-containing bacteria, H. pylori, on the stomach wall. The urea/urease combination creates ammonia which migrates outward through the mucus layer to come into contact with the Reagents.
The urea is added to a soluble, dense carrier at a~ t~ 1-20 grams per kilogram of carrier. The preferred carrier is beads, such as non pareil beads, although any dense carrier can be used which has sufficient density to carry the capsule to the stomach mucosa. In the preferred embodiment the Reagents 1 and 2 are put into the stomach in a solid phasel such as beadsl which can be indi-idually identified in the stomach. The reagents should be coated onto small diameter beads, preferably 0.2-3.0 mm, ~ith the preferred size being a~lt " l~ 2 mm. The 2 mm. srze of the beads pro~ides the advantages of visibility as well as preventing obstruction of the endo-scope in the event not all of the beads dissolve. A suitable method of making such beads would be .
,74~33 to use sugar beads, such as non pareil seeds, with a mesh size of 25-35. The ron pareii beads pro~ide the density required to migrate to the mucosa, either in the capsule or after the capsule dissolves. A less dense vehicle, which iloats within the gastric juices, would prevent the Reagerts from being placed onto the mucosa. U S. Patent No. 3,121,041, issued to Stern et al, discloses the use of a plug, impregnated with a radioactive material, in combination with a soluble capsule.
The spongy plug disclosed in Stern would fioat within the gastric juices, providing several disad-vantages. In order to obtain the contrasting results of the two reagents, two impregnated sponges must be used within the capsule, thereby increasing r ' ;~,~7 expenses. The Stern et al patent discloses tapping the sponges into the capsule. The use of two sponges would possibly double the time required to produce the Stern capsule. Additionally, as the sponges would fioat vithin the gastric juices, the Reagents would be diluted and possibly affected by the contents of the gastric juices. The Reagents must be placed directly onto the mucosa to allow the urea to migrate to the stomach wall, react to the urease created by the H. pylori, create ammonia, and sub-sequently alter the pH. To allow for a dilution factor wouid require increasing the amount of urea used in the capsule. The goal of the instant disclosure, as well as other disclosures relating to the use of radioactive materials, is to reduce the quantity administered to the patient. By placing the urca directly onto the mucosa, dilution is reduced to a minimum and therefore a small quan-tity produces superior accuracy. The beads cannot be coated as commonly known in the time release capsule art, as the reagents on all the beads must be activated ' '~ to obtain a reliable reading. U. S. Patent 3,383,283 to Brindamour discloses time release beads coated with a fatty acid. The fatty acid coating, along with many other coatings, would cause all or some of the beads to float within the gastric juices, again preventing contact with the mucosa.
The disclosed testing procedure is performed in vivo, thereby frequently eliminating the need for a biopsy. In order to view the reagent color change, the beads must remain in a single area. To accomplish this, the beads must not fioat, but rather lie directly on the mucosa, at the source of the bacteria. It has recently been discovered that H. pylori v~ithin the stomach is not continuous or in large areas, but rather patchy within the stomach wall. In the instant disciosure, Wo 95/11672 PCT/US9-ln2332 ~ 9~
the Datural dispersal of ~he beads onto the mucosa cover a suffficient areas to react with at least one area of ~1. pylori bacteria. Any floating indicators which come m contact with the mucosa on either a temporary or scattered basis, bave a narrow chance to come in direct contact with the af-fected area.
Beads which do not dissolve after a few minutes m the stomach can cause an obstruction of the endoscope if they are below the preferred size. Other types of dense vehicles can be used as long as they are capable of absorbing the required reagents and of dissolvirg within a few minutes. When using a powdered carrier, the reagents are mixed with the carrier, the carrier is al-lowed to dry, and, if necessary, re-ground to powder form. The beads have the advantage that coating the beads with the reagents is a simpler, more economical method of obtaining optimum results.
An er~ampie of manufacture of the beads would be:
Reagent I - IJ~ ' blue indicator buffer (pH=6.0) sugar beads Reagent 2 - phenol red indicator buffer (pH=6.0) sugar beads urea The beads are preferably encapsulated into a quick-dissolving gelatin capsule for delivery to the stomach in mass and undiluted. The capsule can be swallowed with a small amount of li-quid, such as water, to more rapidly deliver the capsule and speed the dissol-ing of the capsule. If necessar~, a buffer, such as citrate, ha-ing a pH between 4.0 and 6.0 can be added to the liquid to render the gastrie pH initially slightly acid. Reagents applied in liquid form will mr~ uith each other, even if tal~en separately, providing an indefinite result.
WO 95/llG72 PCT/US9.1/12332 2~74933 Addi~ional ingredien~s can be added with the reagents to produce any specific desired results. An example of this would be to buffer an Acid pH with a stable buffer such as citrate buffer at pH 6.0, 30 mis. The buffer can be added to the seed-coating along with the reagents or can be placed in powdered form in the capsule. The use of a buffer adds stability to the shelf life of the capsules.
In Figure 1 the stomach wall, bacteria with urease, and mucus layers are shown with the reagent beads resting on the mucus layer. As the urea released from the Reagent 2 comes in con-tact with the urease, ammonia is generated. The ammonia rises through the mucus layer and comes into contact with the Reagent indicators, causing an increase in the pH and the Reagents to change color.
To administer the test, the subject takes one to two capsules with 30 mis of pH 6.0 buffer immediately before endoscopy. It takes ~ minutes for the endoscope to reach the stomach, at which time the capsules have dissolved and the granules are resting and slowly dissolv-ing on the surface of the gastric mucosa. Through the endoscope, the examining person can detect the color changes of the reagents, if any, which indicate the presence of the helicobacter or-ganisms.
In the following example Reagent 2 is yellow at acid pH, ehanging to red at alkaline pH
and Reagent I is yellow at acid pH, changing to blue at alkaline pH. The instant invention relies on a differential color change to identify a true positive from a false positive reaction. It is the differential which is of importance, not the colors themselves and any colors and/or reagents specifically used herein are examples and in no way limit the scope of the invention.
READING EXAMPLE I
Nega~ive resul~, (no urease, stomach is acid) Regent 1 (yellow) Both remain yellow no urease Reagent 2 (yellow) no pH change occurs ~ 2 7 ~ 3 g False positive result (stomach has an alkaline pH; for example, bile is in stomach or patient sali-~ates excessively) Reagent 1 (yellow) Changes to blue no ur~cr r'~>6,5 Reagent 2 (yellow) Changes to red READING EXAMPLE III
True posith~e result (stomach is acid but contains urease) Reagent 1 (yellow) urease Remains yellow pH<6 no pH change occurs.
Reagent 2 (yellow) urease Changes red pH rises >65 The presence of red and yellow reagent, but not blue reagent, indicates that urease is in the stomach (i.e. IlLli~ub~
DETECTION OF H. PYL0~I IN THE ST0MACH
BACKGROUND OF THE INVENTION
Brief DescriDtion of the InY~ n~inn The instant invention relates to a novel method of in ivo diagnosis of upper gastromtes-tinal diseases.
Brief DescriDtiQn of the Prior Att Factors adversely affecting the function of the ~ ' ' system in humans are ex-ceedingly varied in their nature, Such disorders may arise in the upper or lower ~ , tracts, or both. There is a broad range of causes of ~ ' disorders, including genetic, physiologicdl, C...;l~ ' and psychogenic hctors. Accordingly, the diagnosis amd manage-ment of these disorders can be ~ difficult.
Among the chronic disorders of the upper t~ ' ' tract are those which fall under the general categories of gastritis and peptic ulcer disease. The upper Er..l~u;.lt~liuldl tract is generally defined as including the esophagus, the stomach, the duodenum, the jejunum and ileum.
Peptic ulcers are lesions of the t~ u;..~c~li..dl tract lining, ~I..,~.lc.i~d by loss of tissue due to the action of digestive acids and pepsin. It has generally been held that peptic ulcers are caused by gastric l~ C~CL;UU~ decreased resistance of the gastric lining to digestive acids and pepsin, or bo~h. Gastritis is, by definition, an ;.31~ ;,." of the stomach mucosa. In practice, though, the disorder is manifested by a broad range of poorly-defined, and heretofore inadequately treated, symptoms such as indigestion, "heart burnn, dyspepsia, and excessive eruaation.
As with the management of any disorder, the rapid, precise, and accurate diagnosis of t,~,.Lr~ ~' disorders is of paramount importance. The typical means used to diagnose the gastrointestinal disorder presented by a given patient v~ill depend upon such factors as the nature and severity of symptoms, the overall health of the individual, the medical history of the patient, the need for a specific diagnosis in order to implement a treatment with reasonable likelihood of success, and the availability of diagnostic devices. However, the diagnostic methods typically employed in the art are often slow, ~ Pr~r~ , costly, and may yield equivocal or inaccurate results. Thus, for patients not ha~ing severe symptoms, a precise diagnosis of a g~l~L-.
-.
~1 7 4933 2 disorder might not be attempted. Sueh patients may simply be treated with eonYentional therapies, sueh as with antaeids or drugs whieh inhibit stomach acid secretion. While such therapies might provide temporary symptomatic relief, a cure is ohen not effected. More effec-tive treatments may depend upon better diagnoses Df the actual underlying ~;callu;~ hlcl disor-der. In particular, it has been discovered that many sueh ~;.laLI- ' ' ' disorders are mediated by infedion of gastrie mucDsa by Helicobaeter pylori. H. pylori is a ( ;rr~ ;cl;._ spiral or-ganism whieh produces the enzyme urease. The organism is ~ / found beneath the mucus layer of the luminal aspect of the gastric epithelium and in the gastrie pits. Helicobader can be diagnosed by blood test for antibodies, breath test, or biopsy of the stomach lining. An-tibodies, however, can remain positive for many months after the baeteria have been eradicated.
The presenee of antibodies presents a falsely positive result in c~ , 10 to 15% of patients. Biopsies are relatively quick; however, they add time, expense and risk. Although rela-ti~ely minor, there is a 1 in 20,000 risk of bleeding from a biopsy site. Biopsies cannot be per-formed on patients who have a tendency to bleed, such as patients with hemophilia and liver dis-ease. Additionally, it has recently been found that helicobacter is patchy, thereby requiring mul-tiple biopsies to obtain 10û% aeeuraey. The eost for a biopsy is C~ ' $100. Biopsies also inerease the risk of the person handling the tissue being exposed to HIV. If a urease test is used, the biopsy sample must be plaeed in the test by the nurse, thereby requiring an additional person during the test.
The prior art has diselosed testing for hcall, ' disorders, the majority of which have been in vitro. Many lests have also been disclosed using urea and indicators.
Marshall, 4,748,113 diseloses eompositions and methods for the diagnosis of ~,calru~ ~lhlcl disorders in-ol-ing urease. Methods inelude obtaining a gastric sample material and eontacting the material with a composition including urease and an indicator.
Marshall 4,830,û10, discloses methods for the diagnosis of ~ lu;~ lcl disorders. The mcthod steps include - ' c~iull of urea-containing compositions prior lo assay.
2 1 7f~933 Steward et al, 5,139,934 disclose substrate ~ and method of urease assay. The method is an in vitro y that includes the use of pH indicators.
Nagatsu et al, 4,147,692 disclose methods and ~ for mcasuring enzymatic ac-tivities and correlating such activities with various disease states, Kraffczyk et al, 3,873,369 disclose colorimetric iDdicators for the .' of urea.
Vasquez et al, 4,851,209 disclose in viYo diagnostic procedures for the clinical evaluation of ' ulcer disease using radioactive isotopes. Procedures involYe prior ' ' ' ~iUI-of a diagnostic ~ l followed by scimigraphic imaging of the ~ area of interest with scintigraphic imaging equipment.
Although the use of urease or other indicators has been used in combination with pH in-dicators, all except Vasquez et al are conducted in vitro.
The instant invention discloses a method of detecting the alkaline pH change in vivo. The test dramatically cnts down the number of biopsies required and is safe for patients having any bleeding tendencies while being rapid and low cost. Addilionally, through the color change, it can be determined if the change is a true positive or a false positive reaction.
BRIEF DESCRIPTION OF THE DRAWINGS
The advantagcs of the instant disclosure will become more apparent when read with the specification and the drâwings, wherein:
FICURE 1 illustra~es the locâtion of the beads in the stomach and the urea/ammonia trans-fer.
SUMMARY OF THE INVENTION
A method for the in vivo detection of urease-producing helicobâcter in the upper stomach is disclosed. A dense carrier is used which is divided into two separate groups, the first combined with a first reagent indicator and the second combined wilh a second reagent indicator and urea, i.e. 14C-Urea or 13C-Urea. The carriers are food soluble products, preferably sugar beads having a diameter of ~ 0.2 to 3.0 mm. The carrier and reagent can be combiDed through coat-ing the carrier or combining the carrier and reagent. The treated carriers and urea are encapsu-.
21 74q33 lated in a soluble capsule which is administered to a patient. A buffer can be added, if desired, toobtain specific results. The density of the carriers cause the capsule to migrate to the g astric mucosa. The gastric juices dissolve the capsule containing the reagents and urea thereby placing the two reagents and urea combination in direct contact with the gastric mucosa. The urea reacts with any urease present in the stomach by creating ammonia, which causes the pH within the stomach to increase. The two reagents react differently, through color change, to the mcrease in pH, which is viewed through use of an endoscope. A preferred first reagent is bromothymol blue (dibrom~h~ . ' ' ' ), which changes yellow in the presence of urease, and a preferred second reagent is phenol red (I ' ' '' , ' ' ' ), which turns red in the presence of urease.
DETAILED DESC~lPrlON OF THE INVENTION
The instant disclosure uses indicators to detect alkaline pH change in the stomach during endoscopy. A change in the colors of the indicators detects pH change within the stomach. A cer-tain combination of colors will indicate the presence of helicobacter, or H. pylori, organisms.
Urea has the formula H2NCONH2 and is a naturally occurring product of protein metabolism. Gastric materials from humans or other animals having t,~ dl disorders contain relatively large quantities of urease (urea . ' ~J~ld~
which hydrolizes urea to ammonium carbonate or ammonia and carbon dioxide. Normally urease is present in the body in only trace amounts, performing the function of ' , ~ urea. H.
pylori increases the amount of urease above normal in the affected areas. The increased urease reacts with the administered urea by creating ammonia, which causes an indicator color change due to the increased alkalinity.
The indicators useful in this invention are weak acids with sharply different colors in their dissociated (ionized) and un-dissociated (neutral) states. The indicators useful herein have PKa values of from about 6.5 to about 8.5, preferably from about 7.0 to about 8Ø The color exhibited by the indicator in the present composition will depend upon the pH of the composition, the particular indicator used, and the dissociation constant (Ka) for that indicator (i.e., PKa ¦ log10Ka). As the color exhibited by the indicator changes over a ~ 74q33 range of pH values (pH=logl0 [H+]), the indicators useful in Ihe present, . chamge color over a pH range of from about 5.5 to about 9.0, preferably from about 6.5 to about 85. The pH of the present composition are accordingly adjusted to a pH at least about one pH unit lower than the PKa of the indicator used (i.e. having a hydrogen ion [H+] ten times less than (10~o of) the hydrogen ion ~ ll in a solution having a pH equal to the pKa of the imdicator).
Preferably, the pH is adjusted to a pH about two pH units below the PKa of the indicato~. Adjust-ment of the pH of the present . can be ef&cted by addition of a base (e.g. sodium hydroxide) or an acid (e.g. hydrochloric acid or citric acid). Thus, preferably, the pH of the oom-position of this invention is adjusted to a pH of from about 5.0 to about 65, with the preferred embodiment being from about 5.0 to about 6Ø
The preferred reagents are 1,.. ' ~ -' blue (d;l,.. ', ' '' . ' ' ' ' ) indicator, Reagent 1, and phenol red (I ' l '' . ' ' ' ) indicator, Reagcnt 2. Other indicators nseful herein include p-nitrophenol, neutral red (2-methyl-3-amino-6-:' ' ,' . ' ), quinoline blue (cyanine), cresol red (o-cr~ ~ , ' ' ' ), and thymol blue (lh, ~ ' ). In-dicators among those useful herein are described in the The Merk Index (9th ed. 1976), incor-porated by reference herein. Reagent 2 has urea added to react to the urease enzyme, if present.
The urea penetrates the mucus layer of the stomach to come into contact with the nrease-containing bacteria, H. pylori, on the stomach wall. The urea/urease combination creates ammonia which migrates outward through the mucus layer to come into contact with the Reagents.
The urea is added to a soluble, dense carrier at a~ t~ 1-20 grams per kilogram of carrier. The preferred carrier is beads, such as non pareil beads, although any dense carrier can be used which has sufficient density to carry the capsule to the stomach mucosa. In the preferred embodiment the Reagents 1 and 2 are put into the stomach in a solid phasel such as beadsl which can be indi-idually identified in the stomach. The reagents should be coated onto small diameter beads, preferably 0.2-3.0 mm, ~ith the preferred size being a~lt " l~ 2 mm. The 2 mm. srze of the beads pro~ides the advantages of visibility as well as preventing obstruction of the endo-scope in the event not all of the beads dissolve. A suitable method of making such beads would be .
,74~33 to use sugar beads, such as non pareil seeds, with a mesh size of 25-35. The ron pareii beads pro~ide the density required to migrate to the mucosa, either in the capsule or after the capsule dissolves. A less dense vehicle, which iloats within the gastric juices, would prevent the Reagerts from being placed onto the mucosa. U S. Patent No. 3,121,041, issued to Stern et al, discloses the use of a plug, impregnated with a radioactive material, in combination with a soluble capsule.
The spongy plug disclosed in Stern would fioat within the gastric juices, providing several disad-vantages. In order to obtain the contrasting results of the two reagents, two impregnated sponges must be used within the capsule, thereby increasing r ' ;~,~7 expenses. The Stern et al patent discloses tapping the sponges into the capsule. The use of two sponges would possibly double the time required to produce the Stern capsule. Additionally, as the sponges would fioat vithin the gastric juices, the Reagents would be diluted and possibly affected by the contents of the gastric juices. The Reagents must be placed directly onto the mucosa to allow the urea to migrate to the stomach wall, react to the urease created by the H. pylori, create ammonia, and sub-sequently alter the pH. To allow for a dilution factor wouid require increasing the amount of urea used in the capsule. The goal of the instant disclosure, as well as other disclosures relating to the use of radioactive materials, is to reduce the quantity administered to the patient. By placing the urca directly onto the mucosa, dilution is reduced to a minimum and therefore a small quan-tity produces superior accuracy. The beads cannot be coated as commonly known in the time release capsule art, as the reagents on all the beads must be activated ' '~ to obtain a reliable reading. U. S. Patent 3,383,283 to Brindamour discloses time release beads coated with a fatty acid. The fatty acid coating, along with many other coatings, would cause all or some of the beads to float within the gastric juices, again preventing contact with the mucosa.
The disclosed testing procedure is performed in vivo, thereby frequently eliminating the need for a biopsy. In order to view the reagent color change, the beads must remain in a single area. To accomplish this, the beads must not fioat, but rather lie directly on the mucosa, at the source of the bacteria. It has recently been discovered that H. pylori v~ithin the stomach is not continuous or in large areas, but rather patchy within the stomach wall. In the instant disciosure, Wo 95/11672 PCT/US9-ln2332 ~ 9~
the Datural dispersal of ~he beads onto the mucosa cover a suffficient areas to react with at least one area of ~1. pylori bacteria. Any floating indicators which come m contact with the mucosa on either a temporary or scattered basis, bave a narrow chance to come in direct contact with the af-fected area.
Beads which do not dissolve after a few minutes m the stomach can cause an obstruction of the endoscope if they are below the preferred size. Other types of dense vehicles can be used as long as they are capable of absorbing the required reagents and of dissolvirg within a few minutes. When using a powdered carrier, the reagents are mixed with the carrier, the carrier is al-lowed to dry, and, if necessary, re-ground to powder form. The beads have the advantage that coating the beads with the reagents is a simpler, more economical method of obtaining optimum results.
An er~ampie of manufacture of the beads would be:
Reagent I - IJ~ ' blue indicator buffer (pH=6.0) sugar beads Reagent 2 - phenol red indicator buffer (pH=6.0) sugar beads urea The beads are preferably encapsulated into a quick-dissolving gelatin capsule for delivery to the stomach in mass and undiluted. The capsule can be swallowed with a small amount of li-quid, such as water, to more rapidly deliver the capsule and speed the dissol-ing of the capsule. If necessar~, a buffer, such as citrate, ha-ing a pH between 4.0 and 6.0 can be added to the liquid to render the gastrie pH initially slightly acid. Reagents applied in liquid form will mr~ uith each other, even if tal~en separately, providing an indefinite result.
WO 95/llG72 PCT/US9.1/12332 2~74933 Addi~ional ingredien~s can be added with the reagents to produce any specific desired results. An example of this would be to buffer an Acid pH with a stable buffer such as citrate buffer at pH 6.0, 30 mis. The buffer can be added to the seed-coating along with the reagents or can be placed in powdered form in the capsule. The use of a buffer adds stability to the shelf life of the capsules.
In Figure 1 the stomach wall, bacteria with urease, and mucus layers are shown with the reagent beads resting on the mucus layer. As the urea released from the Reagent 2 comes in con-tact with the urease, ammonia is generated. The ammonia rises through the mucus layer and comes into contact with the Reagent indicators, causing an increase in the pH and the Reagents to change color.
To administer the test, the subject takes one to two capsules with 30 mis of pH 6.0 buffer immediately before endoscopy. It takes ~ minutes for the endoscope to reach the stomach, at which time the capsules have dissolved and the granules are resting and slowly dissolv-ing on the surface of the gastric mucosa. Through the endoscope, the examining person can detect the color changes of the reagents, if any, which indicate the presence of the helicobacter or-ganisms.
In the following example Reagent 2 is yellow at acid pH, ehanging to red at alkaline pH
and Reagent I is yellow at acid pH, changing to blue at alkaline pH. The instant invention relies on a differential color change to identify a true positive from a false positive reaction. It is the differential which is of importance, not the colors themselves and any colors and/or reagents specifically used herein are examples and in no way limit the scope of the invention.
READING EXAMPLE I
Nega~ive resul~, (no urease, stomach is acid) Regent 1 (yellow) Both remain yellow no urease Reagent 2 (yellow) no pH change occurs ~ 2 7 ~ 3 g False positive result (stomach has an alkaline pH; for example, bile is in stomach or patient sali-~ates excessively) Reagent 1 (yellow) Changes to blue no ur~cr r'~>6,5 Reagent 2 (yellow) Changes to red READING EXAMPLE III
True posith~e result (stomach is acid but contains urease) Reagent 1 (yellow) urease Remains yellow pH<6 no pH change occurs.
Reagent 2 (yellow) urease Changes red pH rises >65 The presence of red and yellow reagent, but not blue reagent, indicates that urease is in the stomach (i.e. IlLli~ub~
Claims (22)
1. The method of in vivo detection of urease-producing helicobacter in the upper stomach comprising the steps of:
obtaining at least two separate groups of dense carriers;
combining the first of said at least two separate groups of dense carriers with a first reagent indicator;
combining the second of said at least two separate groups of dense carriers with a combina-tion of a second reagent indicator and urea;
encapsulating said first reagent and said second reagent-urea combination in a soluble cap-sule;
administering said capsule to a patient;
causing said capsule to migrate to the gastric mucosa through the density of said carriers, dissolving said capsule containing said first reagent and said second reagent-urea combina-tion in the gastric juices;
wherein said first reagent and said second reagent-urea combination are placed in direct contact with the gastric mucosa, allowing said urea to react with any urease present in the stomach thereby creating ammonia, said ammonia causing the pH within said stomach to increase, thereby causing said first reagent and said second reagent to react to said increase in pH, said reac-tion being viewed through endoscopy.
obtaining at least two separate groups of dense carriers;
combining the first of said at least two separate groups of dense carriers with a first reagent indicator;
combining the second of said at least two separate groups of dense carriers with a combina-tion of a second reagent indicator and urea;
encapsulating said first reagent and said second reagent-urea combination in a soluble cap-sule;
administering said capsule to a patient;
causing said capsule to migrate to the gastric mucosa through the density of said carriers, dissolving said capsule containing said first reagent and said second reagent-urea combina-tion in the gastric juices;
wherein said first reagent and said second reagent-urea combination are placed in direct contact with the gastric mucosa, allowing said urea to react with any urease present in the stomach thereby creating ammonia, said ammonia causing the pH within said stomach to increase, thereby causing said first reagent and said second reagent to react to said increase in pH, said reac-tion being viewed through endoscopy.
2. The method of Claim 1 wherein said dense carriers are soluble food products.
3. The method of Claim 2 wherein said food products are sugar beads.
4. The method of Claim 3 wherein said sugar beads have a diameter of approximately 0.2 to 3.0 mm.
5. The method of Claim 1 wherein said first reagent indicator and said second reagent in-dicator react through color change based on the pH present in said stomach.
6. The method of Claim 5 wherein said first reagent and said second reagent change to colors different from one another.
7. The method of Claim 1 wherein said first reagent is a bromothymol blue (dibromothylmolsulfonphthalein) indicator, and said second reagent is a phenol red (phenolsulfonphthalein) indicator.
8. The method of Claim 1 wherein said combination of said dense carrier and said reagent is through coating said carrier with said reagent.
9. The method of Claim 1 wherein said combination of said dense carrier and said reagent is through said carrier absorbing said reagent.
10. The method of Claim 1 wherein said urea is 14C-Urea.
11. The method of Claim 1 wherein said urea is 13C-Urea.
12. The method of Claim 1 wherein a buffer is added to said combination of said dense carrier and said reagents.
13. A diagnostic device for detection of urease-producing helicobacter within the stomach of a human or lower animal subject comprising:
a first dense carrier material, the density of said carrier being sufficient to cause said car-rier to migrate to the gastric mucosa;
a second dense carrier material, the density of said carrier being sufficient to cause said carrier to migrate to the gastric mucosa;
a first reagent, said first reagent being in contact with said first dense carrier, a second reagent, said second reagent being in contact with said second dense carrier;
urea, said urea being in contact with said second dense carrier material;
a soluble capsule, said soluble capsule being soluble in the gastric fluids containing said first carrier and said second carrier.
a first dense carrier material, the density of said carrier being sufficient to cause said car-rier to migrate to the gastric mucosa;
a second dense carrier material, the density of said carrier being sufficient to cause said carrier to migrate to the gastric mucosa;
a first reagent, said first reagent being in contact with said first dense carrier, a second reagent, said second reagent being in contact with said second dense carrier;
urea, said urea being in contact with said second dense carrier material;
a soluble capsule, said soluble capsule being soluble in the gastric fluids containing said first carrier and said second carrier.
14. The diagnostic device of Claim 13 wherein said first dense carrier material and said second dense carrier material are food products.
15. The diagnostic device of Claim 14 wherein said first dense carrier material and said second dense carrier material are beads.
16. The diagnostic device of Claim 13 wherein said contact between said first dense carrier and said first reagent is through coating said carrier with said reagent.
17. The diagnostic device of Claim 13 wherein said contact between said second dense carrier and said second reagent and said urea is through coating said carrier with said reagent and urea.
18. The diagnostic device or Claim 13 wherein said contact between said first dense carrier and said first reagent is through mixing said carrier with said reagent and allowing the combina-tion to dry.
19. The diagnostic device of Claim 13 wherein said contact between said second dense carrier and said second reagent and said urea is through mixing said carrier with said reagent and said urea and allowing the combination to dry.
20. The diagnostic device of Claim 13 wherein said urea is 14C-Urea.
21. The diagnostic device of Claim 13 wherein said urea is 13C-Urea.
22. The method for the diagnosis in a subject of the presence of urease producing helicobacter comprising the steps of:
a- administering to said subject a safe and effective amount of urea, said urea being carried by a dense vehicle, a first portion of said dense vehicle being in combination with a first reagent and a second portion of said dense vehicle being in combination with a second reagent, said vehicle, said reagents and said urea being encapsulated in a capsule, said cap sule being soluble in gastrointestinal fluids, b- drinking approximately 10 ml of water, c- delivering said capsule through gravity to the gastric mucosa, d- dissolving said capsule in the gastric juices thereby allowing said reagent/vehicle combina-tion and said urea to be in direct contact with said gastric mucosa, e- causing the urea to migrate to the stomach wall and reacting with any urease present, wherein said reaction with said urease produces ammonia, thereby altering the pH within the stomach, f- viewing said first reagent and said second reagent through endoscopy for color change, wherein the color change of the first reagent in relation to the color change of the second reagent indicate the presence of urease producing heliobacter within the stomach.
a- administering to said subject a safe and effective amount of urea, said urea being carried by a dense vehicle, a first portion of said dense vehicle being in combination with a first reagent and a second portion of said dense vehicle being in combination with a second reagent, said vehicle, said reagents and said urea being encapsulated in a capsule, said cap sule being soluble in gastrointestinal fluids, b- drinking approximately 10 ml of water, c- delivering said capsule through gravity to the gastric mucosa, d- dissolving said capsule in the gastric juices thereby allowing said reagent/vehicle combina-tion and said urea to be in direct contact with said gastric mucosa, e- causing the urea to migrate to the stomach wall and reacting with any urease present, wherein said reaction with said urease produces ammonia, thereby altering the pH within the stomach, f- viewing said first reagent and said second reagent through endoscopy for color change, wherein the color change of the first reagent in relation to the color change of the second reagent indicate the presence of urease producing heliobacter within the stomach.
Applications Claiming Priority (2)
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|---|---|---|---|
| US14260093A | 1993-10-28 | 1993-10-28 | |
| US08/142,600 | 1993-10-28 |
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| CA2174933A1 true CA2174933A1 (en) | 1995-05-04 |
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| CA002174933A Abandoned CA2174933A1 (en) | 1993-10-28 | 1994-10-25 | Detection of h. pylori in the stomach |
Country Status (10)
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| EP (1) | EP0725633B1 (en) |
| JP (1) | JP3810077B2 (en) |
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| AT (1) | ATE262322T1 (en) |
| AU (1) | AU8127094A (en) |
| BR (1) | BR9407718A (en) |
| CA (1) | CA2174933A1 (en) |
| DE (1) | DE69433646T2 (en) |
| WO (1) | WO1995011672A1 (en) |
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| SE504902C2 (en) * | 1994-11-02 | 1997-05-26 | Diabact Ab | Preparation for detection of Helicobacter pylori in the stomach |
| US6479278B2 (en) * | 1995-06-13 | 2002-11-12 | Barry Marshall | Detection of Helicobacter pylori in the stomach |
| SE9601659D0 (en) * | 1996-04-30 | 1996-04-30 | Diabact Ab | Diagnostic drug preparation |
| US6186958B1 (en) | 1997-02-26 | 2001-02-13 | Oridion Medical | Breath test analyzer |
| US6067989A (en) * | 1997-02-26 | 2000-05-30 | Oridion Medical, Ltd. | Breath test for the diagnosis of Helicobacter pylori infection in the gastrointestinal tract |
| MX9703918A (en) * | 1997-05-28 | 1998-11-30 | J Marshall M D Barry | Procedure to prepare a reactive pharmaceutical product to detect gastrointestinal disorder caused by bacteria in superior gastrointestinal tract. |
| USRE38728E1 (en) * | 1997-09-11 | 2005-04-19 | Oridion Medical, LTD | Breath test analyzer |
| CA2224551A1 (en) * | 1998-02-25 | 1999-08-25 | Alex Hongsheng Chang | A quick in situ diagnostic method for detecting alimentary diseases |
| US20030073155A1 (en) * | 2001-10-15 | 2003-04-17 | Mcmichael Donald J. | Methods for performing multiple diagnostic tests |
| US7008777B2 (en) * | 2001-10-15 | 2006-03-07 | Barry J. Marshall | System for the detection of urease and method for using same |
| US6998250B2 (en) | 2001-10-15 | 2006-02-14 | Donald J. McMichael | Method for detecting Helicobacter pylori |
| US20030072686A1 (en) * | 2001-10-15 | 2003-04-17 | Marshall Barry J. | Systems for performing multiple diagnostic tests |
| US6929926B2 (en) * | 2001-10-15 | 2005-08-16 | Barry J. Marshall | Composition for the detection of gastrointestinal disorders |
| US6783976B2 (en) | 2001-12-21 | 2004-08-31 | Kimberly-Clark Worldwide, Inc. | Carrier and specimen-handling tool for use in diagnostic testing |
| US20060052667A1 (en) * | 2002-10-31 | 2006-03-09 | Yoram Palti | System and method for in vivo detection of h. pylori |
| US20040157281A1 (en) * | 2003-02-11 | 2004-08-12 | Chemsensing, Inc. | Method and apparatus for detecting an analyte |
| JP2004248956A (en) * | 2003-02-21 | 2004-09-09 | Fuji Photo Optical Co Ltd | Pretest capsule for endoscope |
| US7413550B2 (en) * | 2003-10-16 | 2008-08-19 | Kimberly-Clark Worldwide, Inc. | Visual indicating device for bad breath |
| US7582485B2 (en) * | 2003-10-16 | 2009-09-01 | Kimberly-Clark Worldride, Inc. | Method and device for detecting ammonia odors and helicobacter pylori urease infection |
| US6991898B2 (en) | 2003-10-20 | 2006-01-31 | Kimberly-Clark Worldwide, Inc. | Diagnostic test device and method of using same |
| US7577283B2 (en) * | 2005-09-30 | 2009-08-18 | Given Imaging Ltd. | System and method for detecting content in-vivo |
| US7567692B2 (en) * | 2005-09-30 | 2009-07-28 | Given Imaging Ltd. | System and method for detecting content in-vivo |
| US20110092768A1 (en) * | 2007-10-31 | 2011-04-21 | Given Imaging Ltd. | Device, system and method for in-vivo analysis |
| CN106834412A (en) * | 2015-12-03 | 2017-06-13 | 李和楼 | A kind of method of inspection of helicobacter pylori |
| CN106468663B (en) * | 2016-11-11 | 2023-06-13 | 天益健康科学研究院（镇江）有限公司 | Quick detection test bar for helicobacter pylori |
| CN109662693A (en) * | 2018-12-29 | 2019-04-23 | 深圳普门科技股份有限公司 | A kind of miniature detection capsule of Hp based on gas detection |
| CN111289505A (en) * | 2020-03-16 | 2020-06-16 | 南京康容健康科技有限公司 | Helicobacter pylori detect reagent box |
| CN111398511A (en) * | 2020-04-03 | 2020-07-10 | 杭州布封科技有限公司 | Detection device and detection method for detecting helicobacter pylori |
| KR102379748B1 (en) * | 2020-07-13 | 2022-03-29 | 재단법인대구경북과학기술원 | Diagnosing capsule for helicobactor pylori |
| CN114032286A (en) * | 2021-10-27 | 2022-02-11 | 江西省金洹医疗器械股份有限公司 | Helicobacter pylori detection plate and detection method |
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| US3121041A (en) * | 1960-07-20 | 1964-02-11 | Olin Mathieson | Capsule containing a pharmaceutically useful radioactive material |
| US3383283A (en) * | 1964-01-24 | 1968-05-14 | Merck & Co Inc | Medicinal pellets coated with overlapping porous fatty acid leaflet layers |
| ZA733611B (en) * | 1972-10-11 | 1974-04-24 | Merck Patent Gmbh | Indicator for the determination of urea |
| US4748113A (en) * | 1985-06-13 | 1988-05-31 | Marshall Barry J | Compositions and methods for the diagnosis of gastrointestinal disorders involving urease |
| US4830010A (en) * | 1986-04-04 | 1989-05-16 | Marshall Barry J | Methods for the diagnosis of gastrointestinal disorders |
| DD294634A5 (en) * | 1990-05-21 | 1991-10-10 | Zi Fuer Isotopen- Und Strahlenforschung,De | MEDIUM FOR NON-INVASIVE DIAGNOSIS OF HELICOBACTER PYLORI GASTRITIS |
| US5262156A (en) * | 1991-08-12 | 1993-11-16 | Hycor Biomedical, Inc. | Antigenic compositions and their use for the detection of Helicobacter pylori |
| US5314804A (en) * | 1992-03-24 | 1994-05-24 | Serim Research Corporation | Test for Helicobacter pylori |
| US5439801A (en) * | 1994-02-14 | 1995-08-08 | Chek-Med Systems, Inc. | Test composition for the rapid detection of helicobacter pylori in gastric biopsy tissue |
| US5498528A (en) * | 1994-06-10 | 1996-03-12 | King; Wing | Detection of helicobacter pylori |
| US5989840A (en) * | 1997-05-29 | 1999-11-23 | Americare International Diagnostics, Inc. | Estimation of active infection by heliobacter pylori |
-
1994
- 1994-10-25 JP JP51282695A patent/JP3810077B2/en not_active Expired - Fee Related
- 1994-10-25 DE DE69433646T patent/DE69433646T2/en not_active Expired - Fee Related
- 1994-10-25 WO PCT/US1994/012332 patent/WO1995011672A1/en active IP Right Grant
- 1994-10-25 BR BR9407718A patent/BR9407718A/en not_active Application Discontinuation
- 1994-10-25 AU AU81270/94A patent/AU8127094A/en not_active Abandoned
- 1994-10-25 EP EP95900448A patent/EP0725633B1/en not_active Expired - Lifetime
- 1994-10-25 CA CA002174933A patent/CA2174933A1/en not_active Abandoned
- 1994-10-25 AT AT95900448T patent/ATE262322T1/en not_active IP Right Cessation
- 1994-10-25 CN CNB94194624XA patent/CN1137732C/en not_active Expired - Fee Related
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1997
- 1997-03-26 US US08/832,332 patent/US6228605B1/en not_active Expired - Fee Related
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| ATE262322T1 (en) | 2004-04-15 |
| EP0725633A1 (en) | 1996-08-14 |
| CN1139381A (en) | 1997-01-01 |
| JP3810077B2 (en) | 2006-08-16 |
| BR9407718A (en) | 1997-11-11 |
| WO1995011672A1 (en) | 1995-05-04 |
| DE69433646D1 (en) | 2004-04-29 |
| CN1137732C (en) | 2004-02-11 |
| US6228605B1 (en) | 2001-05-08 |
| JPH09506246A (en) | 1997-06-24 |
| AU8127094A (en) | 1995-05-22 |
| EP0725633B1 (en) | 2004-03-24 |
| DE69433646T2 (en) | 2005-02-17 |
| EP0725633A4 (en) | 1998-04-22 |
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