تشخیص ژن اختصاصی eae باکتری انتروپاتوژنیک اشرشیاکلی با استفاده از روش نوین PCR-ELISA

Background and purpose: Enteropathogenic Escherichia coli from Enterobacteriaceae family is one of the most common causes of chronic diarrhea in children and infants. Polymerase Chain Reaction (PCR) method is commonly used for detection of enteropathogenic Escherichia coli species, but there are some disadvantages with this method due to the use of gel electrophoresis and staining with ethidium bromide, including being time consuming, limits on the number of samples, and toxicity of ethidium bromide. The aim of this study was to evaluate the PCR-ELISA technique for detection of enteropathogenic E.coli. Materials and methods: In this study, Streptavidin was loaded in ELISA plate, and a specific Biotinylated probe was used to connect the PCR product. Biotinylated probe was connected to Streptavidin and the amplified gene was attached to the probe. Finally, the digoxigenin antibodies were used to identify the PCR product. The reaction was measured with an ELISA reader. Results: eae of enteropathogenic Escherichia coli was amplified using the gene specific primers which resulted in a fragment of a 999 bp. The results of PCR-ELISA showed that this technique does not cross-react with the bacteria in their families and its sensitivity was 11 pg. Conclusion: PCR-ELISA technique is an accurate and rapid test for detection of infectious agents by the specific gene. PCR-ELISA could be used as an alternative method instead of time-consuming, less sensitive, and expensive techniques.