Rapid screening of toxigenic vibrio cholerae O1 strains from south Iran by PCR-ELISA.
The ability to sensitively detect Vibrio cholera with PCR-ELISA method represents a considerable advancement over alternative more time-consuming methods for detection of this pathogen. The aim of this research is to evaluate the suitability of a PCR-enzyme-linked immunosorbent assay for sensitive and rapid detection of V. cholera O1.



The 398-bp sequence of a gene that codes for the cholera toxin B subunit was amplified by PCR. The digoxigenin-labeled amplified products were coated on microplates and detected by ELISA. The PCR product was also hybridized with biotin labelled probe and detected by ELISA using streptavidin.



The specificity of the PCR was determined using 10 bacterial strains and 50 samples from south Iran. The detection limit was 0.5 pg of the genomic DNA and five bacterial cells. Adaptation of PCR into PCR-ELISA assay format facilitates specific and sensitive detection and diagnosis of human cholera disease. We conclude that this PCR-ELISA is a diagnostic method that specifically detects toxin genes in V. cholera O1 strains. It is more rapid and less cumbersome than other diagnostic methods for detection of toxicity in these strains.
Mousavi
Seyed Latif
SL
Dept. of Biology, College of Basic Sciences, Shahed University, Tehran, Iran. slmousavi@shahed.ac.ir
Nazarian
Shahram
S
Medical Biology Research Center, Baqiyatallah University of Medical Science, Dept. of Biology, Imam Hussein University, Tehran.
Amani
Jafar
J
Medical Biology Research Center, Baqiyatallah University of Medical Science, Dept. of Biology, Imam Hussein University, Tehran.
Rahgerdi
Ahmad Karimi
AK
Malek Ashtar University of Technology, Tehran, Iran.
D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D041241:Vibrio cholerae O1 / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity