Polymerase chain reaction (PCR) for the rapid detection of Salmonella in desiccated coconut (DC)

  • Jayaratne, D.L.
Keywords: Polymerase Chain Reaction (PCR), Salmonella, Desiccated coconut (DC)

Abstract

This study was carried out to develop a polymerase chain reaction (PCR) protocol for the detection of Salmonella in desiccated coconut (DC). For the PCR detection of Salmonella, genomic DNA was extracted using the ‘boiling lyses’ method and the reaction was carried out with Salmonella genus specific primers enabled to amplify 457bp sequence covering invA and invE genes. Samples of DC produced in mills already tested for Salmonella using conventional cultural methods gave identical results with the present PCR method indicating its suitability for adoption in routine testing. 

The sensitivity checked using DNA extracted from artificially inoculated DC with serially diluted inoculum of Salmonella M1 type showed that the developed PCR method can be used to detect very low levels of contamination of Salmonella as low as 4 CFU/g in DC. 

The method described here reduces the testing and detection time from 6 days to 24 hours ensuring exporters to obtain Salmonella test reports just prior to shipment. 

Author Biography

Jayaratne, D.L.

Department of Microbiology, Faculty of Science, University of Kelaniya, Sri Lanka

References

API 20 E Instruction Manual (2000) BioMerieux, Inc. An Identification system for Enterobacteriaceae 20 100/20 160.

CDA, 1972. Microbial method Manual, Coconut Development Authority, Colombo 05, Sri Lanka.

CDA, 2003, Annual report, Coconut Development Authority, Colombo 05, Sri Lanka.

CDA, 2004. Sri Lanka Coconut Statistics, Coconut Development Authority, Colombo 05, Sri Lanka.
.
Galbrraith, S. L., Galan, J. E., Ginocchio, C., Costeas, P and Hedrick, P.,1960. Environment and Public Health. 28.

Guo, L., Killefer, J., Kenney, P. B and Amick-Morris, J. D, 1998. Use of arbitrarily primed polymerase chain reaction to study Salmonella ecology in a turkey production environment. In Environment and Health, Division of Animal and Veterinary Sciences, West Virginia University, Moragantown, west Virginia 26506-61608.

Joseph, M. D., Smedt, S and Amick-Morris, J. D, 1999. Environment and Health 24: 36-38.

Stone, G.G., Oberst, R. D., Hays, M. P., McVey, S and Chengappa, M. M, 1994. Detection of Salmonella serovars from clinical samples by enrichment broth cultivation – PCR procedure. In. Journal of Clinical Microbiology, July 1994: 1742-1749.

Smithsonian Institution (2006). The history of PCR,. Smithsonian Institution Archives, Institutional History Division. Retrieved 24 June 2006.

Wilson, A. K., Black, R., Levine, M. M., Brakette, R. E and Bustler, J. P, 1955. Typhoid fever and infection, Genus Salmonella, In Enterobacteriaceae. 143: 154-186.

Winters, D. K., O’Leary, A. E and Slavik, M. F, 1998. Polymerase chain reaction for rapid detection of Camphylobacter jejuni in artificially contaminated foods. In. Applied Microbiology 27. 163-167.
Published
2007-10-01
How to Cite
Jayaratne, D.L. (2007). Polymerase chain reaction (PCR) for the rapid detection of Salmonella in desiccated coconut (DC). CORD, 23(2), 7. https://doi.org/10.37833/cord.v23i2.165
Section
Articles