Volume 28, Issue 170 (3-2019)
J Mazandaran Univ Med Sci 2019, 28(170): 43-55 |
Back to browse issues page
Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Rami S, Amani J, Saleh T. Detection of Specific eae Gene from Enteropathogenic Escherichia coli by PCR-ELISA. J Mazandaran Univ Med Sci 2019; 28 (170) :43-55
URL: http://jmums.mazums.ac.ir/article-1-11682-en.html
URL: http://jmums.mazums.ac.ir/article-1-11682-en.html
Abstract: (4168 Views)
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.
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.
Type of Study: Research(Original) |
Subject:
Microbiology
Send email to the article author
| Rights and permissions | |
 |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
Articles Copyright © The Author(s). Owned by Mazandaran University of Medical Sciences.
Contact Us
Address: Journal Office, Building No. 2, Mazandaran University of Medical Sciences, Moallem Square, Sari.Tel: +98 (011)34484874 Postal code: 48175-866 E-Mail: jmums@mazums.ac.ir Telefax: +98 (011)33262679
