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Detection of heat-labile toxin in enterotoxigenic Escherichia coli using PCR-ELISA technique
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Esfandiari P1   , Amani J * 2, Imani Fooladi AA3 , Forghanifard MM4 , Mirhossaini SA5 |
1- Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran and Department of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran
2- Associate Professor, Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran , jafar.amani@gmail.com
3- Associate Professor, Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran
4- Assistant Professor, Department of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran
5- Ph.D Candidate in Bacterial Toxin, Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran |
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Abstract: (12874 Views) |
| Background and Objective: Enterotoxigenic Escherichia coli (ETEC) are the most common agent which causes diarrhea, worldwide. ETEC is colonized along the cells and then producing heat-labile (LT) and heat-stable enterotoxigenic which enter into intestinal epithelial cells and causes water and electrolyte loss from intestinal epithelial cells and eventually cause diarrhea.This study was done to detect the heat-labile toxin in Enterotoxigenic Escherichia coli using PCR-ELISA technique. Methods: In this descriptive study, DIG-labeled PCR products were bounded to streptoavidin-coated wells of a microtiter plate and detected by anti-DIG–peroxidase conjugate. The biotin-labeled internal probe was used for verification of PCR products. Results: Heat-labile toxin was detected by PCR-ELISA method. The sensitivity of heat-labile toxin was 1.9 ng. This method did not cross-react with bacteria from this variety. Conclusion: PCR-ELISA method is 100 times more sensitive than conventional PCR method and due to lack of agarose gel and electrophoresis device it can be a good alternative to traditional method. |
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| Keywords: Enterotoxigenic Escherichia coli, PCR-ELISA, Heat-labile enterotoxin |
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Full-Text [PDF 308 kb]
[English Abstract]
(17533 Downloads)
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Type of Study: Original Articles |
Subject:
Microbiology
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