Indirect immunofluorescence test and enzyme-linked immunosorbent assay for detection of Campylobacter pylori

Abstract

An indirect immunofluorescence test (IIF) has been developed for detecting Campylobacter pylori in gastroduodenal biopsies. This test was compared with standard methods of C. pylori diagnosis, namely Gram staining and urease test, in a study population of 226 patients; 121 of the biopsy specimens were cultured for C. pylori as well. C. pylori colonization was detected in 154 of 226 patients (68%) by at least one of these methods (11F, 96%; Gram staining, 78%; urease test, 60%; cultivation, 55%). Serum samples from 191 patients of the study population were screened for circulating antibodies to C. pylori by an indirect enzyme-linked immunosorbent assay with whole, untreated bacteria as antigen. Of these serum specimens, 140 (73%) revealed absorbance readings above the limit of positivity, which was determined as an optical density of >0.35 at 405/620 nm. Of 132 serum specimens, 128 (97%) from patients with C. pylori detected in biopsies, but only 12 (20%) of 59 specimens from those without C. pylori detection showed elevated specific antibody levels. Our data revealed that IIF proved to be the superior rapid, sensitive, and specific diagnostic method. The correlation between microbiological findings and the immune response favors our enzyme-linked immunosorbent assay as an additional tool in C. pylori diagnosis.

Seroepidemiology of Helicobacter pylori infection in Greece

“…It must be noted, however, that in Greece members of the family of a patient are asked to donate their blood. The ELISA with an ultracentrifuged superhate from a whole cell sonic.ate was employed because it is generally accepted that ELISA is a suitable assay for this type of study [17,21,23,26,27] and the antigen preparation has been shown to be the most appropriate [17,20}. SDS-PAGE showed that the protein profile of our preparation (66, 60, 29 and 24 kDa) is similar to that described by Newell [17].…”

Indirect immunofluorescence assay for detection of Helicobacter pylori in human gastric mucosal biopsies

“…Performing the IFA on Formalin-preserved, paraffin-embedded gastric mucosal biopsies allows the evaluation of H. pylori infections in collection biopsies with a high degree of sensitivity and specificity. Indirect immunofluorescence in fresh smears of gastric mucosae has showed a higher sensitivity (96%), but its application in collection samples is not possible (7). The sensitivity, specificity, and reproducibility of our IFA showed that it could be useful as an alternative diagnostic test for H. pylori-associated gastritis.…”

“…Specific diagnosis of H. pylori infection requires culture of the gastric mucosal biopsy in selective media. Indirect immunofluorescence assay (IFA) has been used successfully in the diagnosis of infections caused by fastidious organisms and has proved useful to identify H. pyloii in pure cultures (1) and in gastric mucosal smears (7), but its application for direct diagnosis in routine Formalin-preserved, paraffin-embedded biopsies has not been described.…”

Antibodies Against Helicobacter felis in Sera of Cats and Dogs

“…The specificity of the ELISA was assessed by absorbing serum samples with freshly cultivated cells of C. jejuni and H. pylori. This was performed to avoid possible antigenic cross-reactions with closely related organisms (SCHABER et al, 1989). Additionally sera were absorbed with H. felis.…”