Food Biotechnology, 28:50–62, 2014
Copyright © Taylor & Francis Group, LLC
ISSN: 0890-5436 print / 1532-4249 online
DOI: 10.1080/08905436.2013.870911
Rapid Detection of Salmonella
enterica Subspecies enterica
serovar Typhimurium by Loop
Mediated Isothermal
Amplification (LAMP) Test From
Field Chicken Meat Samples
P . Pavan Kumar
1
, R.K. Agarwal
2
,P . Thomas
2
, B. Sailo
1
,
A. Prasannavadhana
2
, Ashok Kumar
1
, J.L. Kataria
1
,
and D.K. Singh
1
1
Division of Veterinary Public Health, Indian Veterinary Research Institute, Bareilly
City, India
2
Division of Veterinary Bacteriology and Mycology, Indian Veterinary Research
Institute, Bareilly City, India
Considering the importance of Salmonella enterica subsp. enterica serovar
Typhimurium in the foodborne diseases, a Typhimurium specific loop mediated
isothermal amplification (LAMP) test was standardized for its rapid detection in
chicken meat. The Optimum results were obtained at 64
o
C and 70 min tempera-
ture-time combination. The sensitivity of LAMP and PCR were compared with serial
10-fold dilution of the 100 ng of DNA. The LAMP test detected 2 pg DNA per reaction
tube, whereas PCR detected 200 pg DNA per reaction. Therefore, the LAMP test
was considered 100 times more sensitive than the PCR. The specificity of LAMP
and PCR analyzed with six different isolates of non-Salmonella and 22 serovars of
non-Typhimurium. None of these isolates were found positive by both LAMP and PCR.
Twenty-eight pure isolates of Salmonella Typhimurium from diverse sources were also
examined by Typhimurium specific LAMP and were all found positive. Two-hundred
twenty-five field chicken meat samples were screened by cultural, PCR, and LAMP
methods. The LAMP and PCR tests were performed by using DNA isolated from 8 h and
18 h enrichment samples, respectively. Typhimurium specific LAMP was shown to be
in 100% correlated with cultural and PCR methods. However, LAMP test delivered the
results within 26 h without sophisticated equipment while PCR and cultural methods
Address correspondence to P. Pavan Kumar, Division of Veterinary Public Health,
Indian Veterinary Research Institute, Izatnagar, Bareilly (U.P.) 243 122, India; E-mail:
pavaa85@gmail.com