A rapid and specific PCR–ELISA for detecting Salmonella typhi

AUTHORS

Seyed Latif Mousavi 1 , * , Jafar Salimiyan 1 , Ahmad Karimi Rahgerdi 1 , Jafar Amani 1 , Shahram Nazarian 1 , Hassan Ardestani 1

1 Department of Biology, Institute of Basic Sciences, Imam-Hossein University, Tehran, Iran

How to Cite: Mousavi S L, Salimiyan J, Karimi Rahgerdi A, Amani J, Nazarian S, et al. A rapid and specific PCR–ELISA for detecting Salmonella typhi, Arch Clin Infect Dis. 2006 ; 1(3):e93386.

ARTICLE INFORMATION

Archives of Clinical Infectious Diseases: 1 (3); e93386
Published Online: July 30, 2006
Article Type: Research Article
Received: May 12, 2019
Accepted: July 18, 2006

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Abstract

Background: Salmonella continues to be a major food borne pathogen for animals and humans. A sensitive and specific PCR–ELISA technique was developed to detect Salmonella typhi.

Materials and methods: The assay was based on the incorporation of digoxigenin-labeled dUTP and a biotin-labeled primers specific for rfbE gene during PCR amplification. The labeled PCR products were bound to streptoavidin-coated wells of a microtiter plate and detected by ELISA. The specificity of the PCR was determined using 4 strains of Entrobacteriaceae family including Escherichia coli, Klebsiella, Proteus, and Enterobacter and 2 strains of salmonella genus including paratyphi A and entritidis.

Results: Among all the strains, only Salmonella typhi was positive. The PCR-ELISA detecting system was able to increase the sensitivity of assay up to 100 fold, compared with a conventional PCR. The detection limit in PCR-ELISA was 2.5ρg in genomic DNA and 20 cells in direct manner per reaction. The entire procedure took about 100 minutes. For further confirmation of the test, internal biotin labeled probe was designed for rfbE gene and detected with streptavidin.

Conclusion: We have developed a rapid and simple PCR-ELISA protocol suitable for routine analysis of viable Salmonella typhi.

Keywords

PCR-ELISA Rapid detection Salmonella typhi rfbE gene

© 2006, Archives of Clinical Infectious Diseases. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.

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