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A quantitative polymerase chain reaction method for the detection in avian faeces of salmonellas carrying the spvR gene

Published online by Cambridge University Press:  15 May 2009

J. Mahon  and
A. J. Lax
J. Mahon
Affiliation:
AFRC Institute for Animal Health, Compton Laboratory, Compton, Newbury, Berkshire, RG16 0NN, UK
A. J. Lax
Affiliation:
AFRC Institute for Animal Health, Compton Laboratory, Compton, Newbury, Berkshire, RG16 0NN, UK
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Summary

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A quick, semi-quantitative method of detecting Salmonella species which contain the virulence plasmid has been developed using the polymerase chain reaction (PCR). A pair of primers have been synthesized encompassing a 500 bp fragment of the spvR virulence gene. Competitor DNA consisting of the spvR gene with a 94 bp deletion situated between the primer recognition sequences, was cloned into a plasmid vector. Co-amplification of the ‘unknown’ target salmonella DNA with known quantities of competitor DNA in the same reaction tube gave PCR products of 500 and 406 bp respectively. Visual assessment of the ratio of the two products on ethidium bromide stained agarose gels provided an estimate of the approximate number of salmonella cells present in avian faeces.

The technique could be applied to detect quantifiably any non-host DNA in clinical samples if a suitable DNA sequence for primer construction is available.

Type
Research Article
Information
Epidemiology & Infection , Volume 111 , Issue 3 , December 1993 , pp. 455 - 464
Copyright
Copyright © Cambridge University Press 1993

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