Subscribe Now Subscribe Today
Research Article

Rapid Detection of Salmonella in Chicken Meat Using Immunomagnetic Separation, CHROMagar, ELISA and Real-time Polymerase Chain Reaction (RT-PCR)

Ensaf G. Taha, A. Mohamed, K.K. Srivastava and P.G. Reddy
Facebook Twitter Digg Reddit Linkedin StumbleUpon E-mail

The main objective of this study was to standardize and compare rapid methods for the detection of Salmonella in meat samples using Immuno-Magnetic Separation (IMS) followed by culturing in CHROMagar Plus media, Enzyme-Linked Immunosorbent Assay (ELISA) and Real-Time Polymerase Chain Reaction (RT-PCR). Ten-fold dilutions of bacterial suspension (S. typhymurium, ATCC13311) were prepared from the original concentration of 1.6 x 106cfu/ml. Chicken wing samples of 25 g each negative for Salmonella were spiked with six different concentrations of bacteria ranging from 106 to 101. These samples were incubated in buffered peptone water for 4 h as pre-enrichment step and were tested repeatedly. The IMS technique involved the use of paramagnetic polystyrene microscopic beads coated with purified antibodies against Salmonella. The CHROMagar Plus media containing chromomeric substrate facilitated detection of Salmonella species from other flora. The Assurance EIA Salmonella Kit with polyclonal antibodies directed against Salmonella facilitated easy and rapid detection. In the RT-PCR primers targeting invA gene was used which amplified a 378 bp fragment. Comparing to conventional culture method (4 days), CHROMagar plate culture following IMS showed light mauve to mauve-colored colonies of Salmonella in 23 h with high sensitivity (99%) at 1.6 cfu/ml. IMS-ELISA combination also showed high sensitivity (75%) at 1.6 x 103 cfu/ml in 8 h and minimized cross-reactivity with many Enterobacteraceae. The combination of IMS with RT-PCR took less than 7 h and was even more sensitive (100%) at 1.6 cfu/ml. Sensitivities of IMS-RT-PCR and IMS-CHROMagar were higher compared to IMS-ELISA. IMS-CHROMagar was easier to perform and detects only living Salmonella. These methods will be highly suitable for routine detection and may significantly assist the processing industry in avoiding costly recalls and the timely investigation of outbreaks.

Related Articles in ASCI
Search in Google Scholar
View Citation
Report Citation

  How to cite this article:

Ensaf G. Taha, A. Mohamed, K.K. Srivastava and P.G. Reddy, 2010. Rapid Detection of Salmonella in Chicken Meat Using Immunomagnetic Separation, CHROMagar, ELISA and Real-time Polymerase Chain Reaction (RT-PCR). International Journal of Poultry Science, 9: 831-835.

DOI: 10.3923/ijps.2010.831.835



1:  Duncanson, P., D.R. Wareing and O. Jones, 2003. Application of an automated immunomagnetic separation-enzyme immunoassay for the detection of Salmonella spp during an outbreak associated with a retail premises. Lett. Applied Microbiol., 37: 144-148.
PubMed  |  

2:  Galan, J.E., C. Ginocchio and P. Costeas, 1992. Molecular and functional characterization of the Salmonella invasion gene i3vA: Homology of invA to members of a new protein family. J. Bacteriol., 174: 4338-4349.
Direct Link  |  

3:  Hara-Kudo, Y., S. Kumagai, T. Masuda, K. Goto and K. Ohtsuka et al., 2001. Detection of Salmonella enteritidis in shell and liquid eggs using enrichment and plating. Int. J. Food Microbiol., 64: 395-399.
CrossRef  |  

4:  Maddocks, S., T. Olma and S. Chen, 2002. Comparison of CHROMagar Salmonella medium and xylose-lysine-desoxycholate and Salmonella-Shigella agar for isolation of Salmonella strains from stool samples. J. Clin. Microbiol., 40: 2999-3003.
PubMed  |  

5:  Mercanoglu, B. and M.W. Griffiths, 2005. Combination of immunomagnetic separation with real-time pcr for rapid detection of Salmonella in milk, ground beef and alfalfa sprouts. J. Food Prot., 68: 557-561.
PubMed  |  

6:  Nam, H.M., V. Srinivasan, B.E. Gillespie, S.E. Murinda and S.P. Oliver, 2005. Application of SYBR green real-time PCR assay for specific detection of Salmonella spp. in dairy farm environmental samples. Int. J. Food Microbiol., 102: 161-171.
PubMed  |  

©  2021 Science Alert. All Rights Reserved