2019 ; Vol.29-3: 454~464
|Author||Zhi Ke Liu, Qiu Yu Zhang, Ning-Ning Yang, Ming-Guo Xu, Jin-Feng Xu, Ming-Long Jing, Wen-Xing Wu, Ya-Dong Lu, Feng Shi, Chuang-Fu Chen|
|Place of duty||College of Animal Science and Technology, Shihezi University, China|
|Title||Rapid and Sensitive Detection of Salmonella in Chickens Using Loop-Mediated Isothermal Amplification Combined with a Lateral Flow Dipstick|
J. Microbiol. Biotechnol.2019 ;
|Abstract||Salmonellosis is a highly contagious bacterial disease that threatens both human and poultry
health. Tests that can detect Salmonella in the field are urgently required to facilitate disease
control and for epidemiological investigations. Here, we combined loop-mediated isothermal
amplification (LAMP) with a chromatographic lateral flow dipstick (LFD) to rapidly and
accurately detect Salmonella. LAMP primers were designed to target the Salmonella invA gene.
LAMP conditions were optimized by adjusting the ratio of inner to outer primers, MgSO4
concentration, dNTP mix concentration, amplification temperature, and amplification time.
We evaluated the specificity of our novel LAMP-LFD method using six Salmonella species and
six related non-Salmonella strains. All six of the Salmonella strains, but none of the non-
Salmonella strains, were amplified. LAMP-LFD was sensitive enough to detect concentrations
of Salmonella enterica subsp. enterica serovar Pullorum genomic DNA as low as 89 fg/μl, which
is 1,000 times more sensitive than conventional PCR. When artificially contaminated feed
samples were analyzed, LAMP-LFD was also more sensitive than PCR. Finally, LAMP-LFD
gave no false positives across 350 chicken anal swabs. Therefore, our novel LAMP-LFD assay
was highly sensitive, specific, convenient, and fast, making it a valuable tool for the early
diagnosis and monitoring of Salmonella infection in chickens.|
|Key_word||Salmonella, invA gene, loop-mediated isothermal amplification, lateral flow dipstick, detection|
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