Evaluation of an enzyme immunoassay for verotoxin detection in Escherichia coli.
Verotoxin-producing Escherichia coli strains (VTEC) cause hemorrhagic colitis and hemolytic-uremic syndrome in humans. Laboratory diagnosis by conventional methods is slow and cumbersome. The results of a new rapid enzyme immunoassay (EIA Premier EHEC) for verotoxin detection both in isolated strains and in clinical samples are presented, and they are compared with cell culture (CC) and polymerase chain reaction (PCR) techniques. Fifty-four strains have been analyzed by both EIA and PCR, and 33 by all three methods. The kit has also been evaluated for experimentally infected stool samples directly and after their enrichment on MacConkey broth. Nineteen, out of the 54 strains, were positive by EIA and 20 by PCR. The results of the 33 strains evaluated by the three techniques were coincident with one exception. The latter was uninterpretable by CC, negative by EIA and positive by PCR. The sensitivity of the kit for experimentally infected stool samples was approximately 5 x 10(7) bacteria/ml in the direct test, and 5 x 10(4) bacteria/ml after broth enrichment. EIA sensitivity and specificity were similar to those of CC and PCR. The diagnostic times were 18h for EIA, 3 days for PCR and 5 days for CC. Sensitivity, rapidity and ease of performance make this technique especially valuable for clinical diagnosis.
Servei de Microbiologia, Hospital de la Santa Creu i Sant Pau, Barcelona, España.
C
C
Frias
M
M
Majò
N
N
Margall
T
T
Llobet
B
B
Mirelis
G
G
Prats
D000818:Animals; D001427:Bacterial Toxins / Q000032:analysis* / Q000235:genetics; D001431:Bacteriological Techniques; D002522:Chlorocebus aethiops; D004269:DNA, Bacterial / Q000032:analysis; D003967:Diarrhea / Q000382:microbiology; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D004927:Escherichia coli Infections / Q000382:microbiology; D005069:Evaluation Studies as Topic; D005243:Feces / Q000382:microbiology; D005798:Genes, Bacterial; D006801:Humans; D007124:Immunoenzyme Techniques; D016133:Polymerase Chain Reaction; D011933:Reagent Kits, Diagnostic; D012680:Sensitivity and Specificity; D022622:Shiga Toxin 1; D022641:Shiga Toxin 2; D013997:Time Factors; D014709:Vero Cells / Q000382:microbiology
D001427:Bacterial Toxins; D004269:DNA, Bacterial; D011933:Reagent Kits, Diagnostic; D022622:Shiga Toxin 1; D022641:Shiga Toxin 2