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Microbiology: Current Research 2017
Volume 1 Issue 2 Microbes Infection 2017
Notes:
Page 41
September 28-29, 2017 | London, UK
Microbes Infection
38th Annual congress on
REAL-TIME PCR METHOD FOR DETECTION
OF SALMONELLA SPP. IN ENVIRONMENTAL
SAMPLES
Kuppuswamy N Kasturi1 and Tomas Drgon2
1US Food and Drug Administration, New York, USA
2US Food and Drug Administration, Maryland, USA
The methods currently used in FDA (Food and Drug
Administration) field laboratories and other public health
laboratories for detecting Salmonella in food/environmental
samples require 2 days and have limited sensitivity. We
describe the development and validation of a real-time PCR
method that detected Salmonella and presence of group
D in 24 h. Primers and probes specific to the invA gene of
Salmonella, group D, and Enteritidis serovar were designed
and evaluated for the inclusivity and exclusivity using a panel
of 329 Salmonella isolates consisting 126 serovars from 32-
O groups and 22 non-Salmonella environmental organisms.
The invA-, group D- and Enteritidis-specific sets identified
the isolates accurately. The PCR method was100% inclusive
for Salmonella spp. and had a detection limit of 2 copies of
Salmonella DNA per reaction. A Single-laboratory validation
performed on 1,741 environmental samples demonstrated
that the PCR method detected 55% more positives than the
Vitek immunodiagnostic assay system method (VIDAS)
method that is currently used. The method is more specific
and did not report any false-negatives. The receiver operating
characteristic (ROC) analysis documented excellent
agreement between the results from the culture and PCR
methods (area under the curve, 0.90; 95% confidence interval
of 0.76 to 1.0) confirming the validity of the PCR method.
The validated PCR method will help to strengthen public
health efforts through rapid screening of Salmonella spp. in
environmental samples.
Microbiology: Current Research 2017