Serological detection of Helicobacter pylori by a flow microsphere immunofluorescence assay.
A flow cytometric immunofluorescence assay (FMIA) for the detection of immunoglobulin G antibodies to Helicobacter pylori was developed. A multicomponent antigen was prepared and used to coat carboxylated polystyrene microspheres for reaction with patient sera followed by fluorescein isothiocyanate-labelled goat anti-human immunoglobulin G. The reacted microspheres were collected with a flow cytometer, and fluorescence was quantitated relative to the cutoff value provided by pooled sera from patients in whom H. pylori could not be demonstrated by culture or histology. Serum samples from 28 H. pylori-positive patients and 27 H. pylori-negative patients were tested by FMIA. Additionally, an in-house enzyme-linked immunosorbent assay (ELISA) employing the same antigen preparation and a commercially available ELISA were used to assay the patient population. Both the FMIA and in-house ELISA were 100% sensitive and 89% specific with positive and negative predictive values of 90 and 100% and no equivocal results. The commercial ELISA was 96% sensitive and 89% specific with positive and negative predictive values of 90 and 96% and five equivocal results. FMIA provides a rapid, inexpensive, and easily performed means for serodiagnosis of H. pylori.
Best
L M
LM
Department of Microbiology, Victoria General Hospital, Halifax, Nova Scotia, Canada.
Veldhuyzen van Zanten
S J
SJ
Bezanson
G S
GS
Haldane
D J
DJ
Malatjalian
D A
DA
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000907:Antibodies, Bacterial / Q000032:analysis*; D000942:Antigens, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D001706:Biopsy; D004797:Enzyme-Linked Immunosorbent Assay; D005069:Evaluation Studies as Topic; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D016481:Helicobacter Infections / Q000175:diagnosis*; D016480:Helicobacter pylori / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D007413:Intestinal Mucosa / Q000382:microbiology; D008297:Male; D008863:Microspheres; D008875:Middle Aged; D011237:Predictive Value of Tests; D011706:Pyloric Antrum / Q000382:microbiology; D011933:Reagent Kits, Diagnostic; D012698:Serologic Tests / Q000379:methods*
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D007074:Immunoglobulin G; D011933:Reagent Kits, Diagnostic