- Email: [email protected]
Evaluation of flexsureTM HP, a rapid serological assay for H. pylori detection
April 1995
Esophageal, Gastric, and Duodenal Disorders
RELATIONSHIP BETWEEN THE MAGNITUDE OF [~3CIUREA BREATH TEST RESULTS AND ENDOSCOPIC DIAGNOSES IN HELICOBACTER PYLORI INFECTED INDIVIDUALS. T.K.Sharma & A.F.Cutler, Section of Gastroenterology, Sinai Hospital, Detroit, MI. Backvound: H. pylori (HP) is an accepted cause of chronic active type B gastritis and plays a significant role in the etiopathogenesis of peptic ulcer disease (PUD). l~3C}urea breath test (UBT) has a high (9499%) sensitivity and specificity for detecting HP status. iAim: To test if a relationship exists between the magnitude of l'3C]urea breath test results and endoscopic findings in HP infected individuals. Methods: Patients who bad undergone endoscopy and had a UBT __> 6% were entered into this retrospective study. They were grouped by endoscopic diagnosis: PUD (DU or GU); non-ulcer dyspepsia (NUD) and other. NUD was defined as dyspeptic symptoms of one month duration with normal upper endoscopy. Mean values were compared between groups (PUD vs no PUD, PUD vs NUD, DU vs GU) using the students T-test (2 tail) with probability of < 5% being significant. Results: 168 patients were entered into the study. The mean age (yrs), gender (%male), mean UBT values were determined:
N Age %Male UBT
Total
PUD
168 55.0 57 25.43
109 54.7 63 24.78
DU
GU
NUD
Other
73 52.6 65 23.74
36 58.9 58 26.90
35 52.2 37 26.36
24 61.3 58 26.99
The FlexSure T M HP is a new bi-directional immunochromatographic device developed by SmithKline Diagnostics, Inc. for the detection of IgG antibodies to H. pylori. It uses the same antigen as the HM-CAP T M ELk kit (Enteric Products, Inc). The FlexSure TM HP is a two step assay with a rapid (4 minutes) incubation step designed to provide an immediate serology result in an office setting. The efficacy of this assay was evaluated in eighty patients with dyspeptic symptoms. All underwent endoscopy, and both HM-CAP EIAT M (the evaluation standard) and FlexSure~HP serologies. Gastric biopsies were examined histologically and tested for urease activity by the CLO test. Histology was not available for nine patients. None had previot.s treatment for H. pylari or recent antibiotic usage.
I+1
f+ I
Hist°l°i~
Linscheer, R. Oates. Veterans Affairs Medical Canter and SUNS~Health Science Center, Syracuse,NY. Exaggerated gaslrin responses in HP gastritis occur without an increase in gaslrin-secrefing G-cells, but with a reciprocal decrease in antral somatostatinsecretingD-cellsand antral somatostatincontent. Activity of G- and D-cells may be reflected by the morphology of their secretory granules, "empty" granules predon,maateinactively secreting cells. In this ultrastsuctural study of antral G- and D-calls we compared secretory-granulemorphologyin 7 HP-infected subjects and 7 HP-negalivecantrols. Methods: HP status was determinedby serology, CLO test and Giemsa staining of antral biopsies from subjects with nonulcer dyspepsia. In a blinded fashion, G- and D-cells were identified by transmissionelectronmicroscopy and photographed. Outlinesof the cell, nucleusand secretory granules were digitized and areas computed on an image analyzer (Zidus, Leitz). Granules were classified as 'Empty','Intermediate'and 'Full' based on their content of electron-dense secretory material. The total granuleand subtype granule, density and fraction were compared in the I-IP+and HP- groups by unpaired t-tests using natural In transformationwhen necessary to control for variability. ~ : Total granule number and density in G-cells was similar in the two groups but there were differences in granule subtype. "Full" granules were reduced inthe HP+ group with a concomitant increase in "empty"granules(see table). Therewereno differencesin D-cell granule number and morphology.
HP+ n=18 HP- n=17
• EVALUATION OF FLEXSURE T M HP, A RAPID SEROLOGICAL ASSAY FOR H. PYLORI DETECTION. R. Shaw, M. Garcia, E. Cheng, P. Marchildon, and J. Peacock. Veterans Affairs Medical Center, Northport, N.Y. 11768, and Enteric Products, Inc., Stony Brook, N.Y. 11794.
I HM-CAP ELk I FlexSure HP I CLO (2 sites) I
ULTRASTRUCTURAL CHANGES IN ANTRAL ENDOCRINE CELLS ]]31
HELICOBACTER PYLORI (liP) GASTRITIS. V.K. Sharma, U.K. Murthy, W.G.
G Cells
Statistical analysis did not reveal any relationship between quantitative UBT results and endoscopic diagnoses. Conclusion: There is no relationship between the magnitude of 1~3C]urea breath test results and the endoscopic diagnoses in HP infected patients.The magnitude of the 1~3C]urea breath test can not be used to predict the presence or absence of peptic ulcer disease.
[
I+1-1
The results show an excellent correlation between the FlexSure T M HP and HM-CAP T M ELk with a specificity of 97% (32/33) and a sensitivity of 88.6% (39/44). CLO and histology positives were all positive by both serologies, although several serology-positive patients had negative CLO and histology. Biopsy-based diagnostics are known to be less senstive due to early infection, recent eradication, reduction in IT. pylarl by unreported antibiotic or omeprazole exposure, or inadequate sampling of gastric mucosa due to the patchy distribution of the bacteria in the stomach. This rapid diagnostic format will enable immediate correlation of H. pylori serology with the presence of gastric and duodenal ulcers, facilitating rational use of antibiotic therapy.
A217
Subtvoe Granule Density
Subtype Granule Fraction
Empty Full Empty Pull 2.13±0.3 1.20±0.2" 38.6+4.5 18.9±1.9" 1.80~0.3 2.15+0.4 28.1±4.4 29.7±3.9 Data = ~ ± SE, *p < 0.05 fer HP+ vs HP-. Dansity=No. of granules/mm2area of cytoplasm. Fraction=subtypegranule No./total granule No. X 100.
Conclusions: Morphologicalchanges in antral G cells in HP gastritis are suggestive of hyperfunction. D-cell morphology remains unchanged, suggestingthat decreased D-ceU number rather than D-cell hypofunctionis the cause of G-cell hyperfunction and exaggerated gastrin response in I-IPgastritis. This study was supported in part by GLAXO Pharmaceuticals.
P H O S P H A T I D Y L C H O L I N E H Y D R O P E R O X I D E I N GASTRIC M U C O S A L L E S I O N S I N D U C E D BY HC1, N a O H OR E T H A N O L : DETECTION USING LUMINOL CHEMILUMINESCENCEH I G H P E R F O R M A N C E L I Q U I D CHROMATOGRAPHY. WeiMing Shian, I. Sasaki, H. Naito, T. Tsuchiya, S. Matsuno. 1st Dept. of Surgery, School of Medicine, Tohoku University, Sendal, J a p a n . Free radicals are believed to be involved in gastric muoosal lesions induced by HC1, N a O H or ethanol. Phosphatidylcholine, the m o s t i m p o r t a n t functional lipid in the cellular m e m b r a n e , is found to c o n s t i t u t e a b o u t h a l f of t h e total phospholipids in gastric m u c o s a , w h i c h is p a r t i c u l a r l y sensitive to peroxidative stress. However, owing to t h e l i m i t a t i o n s of c u r r e n t m e t h o d s , lipid hydroperoxides in biological t i s s u e s h a v e n o t b e e n determined quantitatively, therefore t h e p a t h o g e n e s i s of g a s t r i c m u c o s a l lesions is still obscure. A i m : This s t u d y is designed to determine w h e t h e r phosphatidylcholine hydroperoxide (PCOOH), a p r i m a r y peroxidative product of phosphatidylcholine play a n i m p o r t a n t role in t h e p a t h o g e n e s i s of gastric m u c o s a l lesions induced by HC1, N a O H or ethanol. M e t h o d s : Male Sprague-Dawley r a t s weighing a p p r o x i m a t e l y 200g w e r e s t a r v e d for 24 h o u r s . T h e y were a n e s t h e t i z e d , 30 s, 1, 2, 3, or 15 m i n a f t e r intragastric administration of 0.6 N HC1, 0.2 N N a O H or 60% ethanol (lml), their s t o m a c h s w e r e excised. The levels of P C O O H in gastric m u c o s a w e r e m e a s u r e d b y M i y a z a w a ' s m e t h o d (CL-HPLC). R e s u l t s : Mucosal lesions were visible at 30 s after administration of all t h e necrotizing a g e n t s , a n d i n c r e a s e d t i m e dependently. P C O O H in gastric m u c o s a were significantly elevated at 30 s (P < 0.01, Fisher's PLSD), and the level of P C O O H w a s m a x i m u m at 3 rain, t h e n declined. C o n c l u s i o n : Lipid peroxidation m a y p l a y a crucial role in the p a t h o g e n e s i s of gastric mucosal lesions induced by HC1, N a O H or ethanol. The P C O O H level m a y be a n early detectable index for gastric mucosal lesions.
Esophageal, Gastric, and Duodenal Disorders
RELATIONSHIP BETWEEN THE MAGNITUDE OF [~3CIUREA BREATH TEST RESULTS AND ENDOSCOPIC DIAGNOSES IN HELICOBACTER PYLORI INFECTED INDIVIDUALS. T.K.Sharma & A.F.Cutler, Section of Gastroenterology, Sinai Hospital, Detroit, MI. Backvound: H. pylori (HP) is an accepted cause of chronic active type B gastritis and plays a significant role in the etiopathogenesis of peptic ulcer disease (PUD). l~3C}urea breath test (UBT) has a high (9499%) sensitivity and specificity for detecting HP status. iAim: To test if a relationship exists between the magnitude of l'3C]urea breath test results and endoscopic findings in HP infected individuals. Methods: Patients who bad undergone endoscopy and had a UBT __> 6% were entered into this retrospective study. They were grouped by endoscopic diagnosis: PUD (DU or GU); non-ulcer dyspepsia (NUD) and other. NUD was defined as dyspeptic symptoms of one month duration with normal upper endoscopy. Mean values were compared between groups (PUD vs no PUD, PUD vs NUD, DU vs GU) using the students T-test (2 tail) with probability of < 5% being significant. Results: 168 patients were entered into the study. The mean age (yrs), gender (%male), mean UBT values were determined:
N Age %Male UBT
Total
PUD
168 55.0 57 25.43
109 54.7 63 24.78
DU
GU
NUD
Other
73 52.6 65 23.74
36 58.9 58 26.90
35 52.2 37 26.36
24 61.3 58 26.99
The FlexSure T M HP is a new bi-directional immunochromatographic device developed by SmithKline Diagnostics, Inc. for the detection of IgG antibodies to H. pylori. It uses the same antigen as the HM-CAP T M ELk kit (Enteric Products, Inc). The FlexSure TM HP is a two step assay with a rapid (4 minutes) incubation step designed to provide an immediate serology result in an office setting. The efficacy of this assay was evaluated in eighty patients with dyspeptic symptoms. All underwent endoscopy, and both HM-CAP EIAT M (the evaluation standard) and FlexSure~HP serologies. Gastric biopsies were examined histologically and tested for urease activity by the CLO test. Histology was not available for nine patients. None had previot.s treatment for H. pylari or recent antibiotic usage.
I+1
f+ I
Hist°l°i~
Linscheer, R. Oates. Veterans Affairs Medical Canter and SUNS~Health Science Center, Syracuse,NY. Exaggerated gaslrin responses in HP gastritis occur without an increase in gaslrin-secrefing G-cells, but with a reciprocal decrease in antral somatostatinsecretingD-cellsand antral somatostatincontent. Activity of G- and D-cells may be reflected by the morphology of their secretory granules, "empty" granules predon,maateinactively secreting cells. In this ultrastsuctural study of antral G- and D-calls we compared secretory-granulemorphologyin 7 HP-infected subjects and 7 HP-negalivecantrols. Methods: HP status was determinedby serology, CLO test and Giemsa staining of antral biopsies from subjects with nonulcer dyspepsia. In a blinded fashion, G- and D-cells were identified by transmissionelectronmicroscopy and photographed. Outlinesof the cell, nucleusand secretory granules were digitized and areas computed on an image analyzer (Zidus, Leitz). Granules were classified as 'Empty','Intermediate'and 'Full' based on their content of electron-dense secretory material. The total granuleand subtype granule, density and fraction were compared in the I-IP+and HP- groups by unpaired t-tests using natural In transformationwhen necessary to control for variability. ~ : Total granule number and density in G-cells was similar in the two groups but there were differences in granule subtype. "Full" granules were reduced inthe HP+ group with a concomitant increase in "empty"granules(see table). Therewereno differencesin D-cell granule number and morphology.
HP+ n=18 HP- n=17
• EVALUATION OF FLEXSURE T M HP, A RAPID SEROLOGICAL ASSAY FOR H. PYLORI DETECTION. R. Shaw, M. Garcia, E. Cheng, P. Marchildon, and J. Peacock. Veterans Affairs Medical Center, Northport, N.Y. 11768, and Enteric Products, Inc., Stony Brook, N.Y. 11794.
I HM-CAP ELk I FlexSure HP I CLO (2 sites) I
ULTRASTRUCTURAL CHANGES IN ANTRAL ENDOCRINE CELLS ]]31
HELICOBACTER PYLORI (liP) GASTRITIS. V.K. Sharma, U.K. Murthy, W.G.
G Cells
Statistical analysis did not reveal any relationship between quantitative UBT results and endoscopic diagnoses. Conclusion: There is no relationship between the magnitude of 1~3C]urea breath test results and the endoscopic diagnoses in HP infected patients.The magnitude of the 1~3C]urea breath test can not be used to predict the presence or absence of peptic ulcer disease.
[
I+1-1
The results show an excellent correlation between the FlexSure T M HP and HM-CAP T M ELk with a specificity of 97% (32/33) and a sensitivity of 88.6% (39/44). CLO and histology positives were all positive by both serologies, although several serology-positive patients had negative CLO and histology. Biopsy-based diagnostics are known to be less senstive due to early infection, recent eradication, reduction in IT. pylarl by unreported antibiotic or omeprazole exposure, or inadequate sampling of gastric mucosa due to the patchy distribution of the bacteria in the stomach. This rapid diagnostic format will enable immediate correlation of H. pylori serology with the presence of gastric and duodenal ulcers, facilitating rational use of antibiotic therapy.
A217
Subtvoe Granule Density
Subtype Granule Fraction
Empty Full Empty Pull 2.13±0.3 1.20±0.2" 38.6+4.5 18.9±1.9" 1.80~0.3 2.15+0.4 28.1±4.4 29.7±3.9 Data = ~ ± SE, *p < 0.05 fer HP+ vs HP-. Dansity=No. of granules/mm2area of cytoplasm. Fraction=subtypegranule No./total granule No. X 100.
Conclusions: Morphologicalchanges in antral G cells in HP gastritis are suggestive of hyperfunction. D-cell morphology remains unchanged, suggestingthat decreased D-ceU number rather than D-cell hypofunctionis the cause of G-cell hyperfunction and exaggerated gastrin response in I-IPgastritis. This study was supported in part by GLAXO Pharmaceuticals.
P H O S P H A T I D Y L C H O L I N E H Y D R O P E R O X I D E I N GASTRIC M U C O S A L L E S I O N S I N D U C E D BY HC1, N a O H OR E T H A N O L : DETECTION USING LUMINOL CHEMILUMINESCENCEH I G H P E R F O R M A N C E L I Q U I D CHROMATOGRAPHY. WeiMing Shian, I. Sasaki, H. Naito, T. Tsuchiya, S. Matsuno. 1st Dept. of Surgery, School of Medicine, Tohoku University, Sendal, J a p a n . Free radicals are believed to be involved in gastric muoosal lesions induced by HC1, N a O H or ethanol. Phosphatidylcholine, the m o s t i m p o r t a n t functional lipid in the cellular m e m b r a n e , is found to c o n s t i t u t e a b o u t h a l f of t h e total phospholipids in gastric m u c o s a , w h i c h is p a r t i c u l a r l y sensitive to peroxidative stress. However, owing to t h e l i m i t a t i o n s of c u r r e n t m e t h o d s , lipid hydroperoxides in biological t i s s u e s h a v e n o t b e e n determined quantitatively, therefore t h e p a t h o g e n e s i s of g a s t r i c m u c o s a l lesions is still obscure. A i m : This s t u d y is designed to determine w h e t h e r phosphatidylcholine hydroperoxide (PCOOH), a p r i m a r y peroxidative product of phosphatidylcholine play a n i m p o r t a n t role in t h e p a t h o g e n e s i s of gastric m u c o s a l lesions induced by HC1, N a O H or ethanol. M e t h o d s : Male Sprague-Dawley r a t s weighing a p p r o x i m a t e l y 200g w e r e s t a r v e d for 24 h o u r s . T h e y were a n e s t h e t i z e d , 30 s, 1, 2, 3, or 15 m i n a f t e r intragastric administration of 0.6 N HC1, 0.2 N N a O H or 60% ethanol (lml), their s t o m a c h s w e r e excised. The levels of P C O O H in gastric m u c o s a w e r e m e a s u r e d b y M i y a z a w a ' s m e t h o d (CL-HPLC). R e s u l t s : Mucosal lesions were visible at 30 s after administration of all t h e necrotizing a g e n t s , a n d i n c r e a s e d t i m e dependently. P C O O H in gastric m u c o s a were significantly elevated at 30 s (P < 0.01, Fisher's PLSD), and the level of P C O O H w a s m a x i m u m at 3 rain, t h e n declined. C o n c l u s i o n : Lipid peroxidation m a y p l a y a crucial role in the p a t h o g e n e s i s of gastric mucosal lesions induced by HC1, N a O H or ethanol. The P C O O H level m a y be a n early detectable index for gastric mucosal lesions.