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Robustness of S almonella loop‐mediated isothermal amplification assays for food applications
Author(s) -
Yang Q.,
Wang F.,
Prinyawiwatkul W.,
Ge B.
Publication year - 2014
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.12340
Subject(s) - loop mediated isothermal amplification , salmonella , robustness (evolution) , microbiology and biotechnology , biology , biological system , computational biology , bacteria , genetics , gene , dna
Aims Loop‐mediated isothermal amplification ( LAMP ) assays have been developed recently for S almonella detection. This study aimed at evaluating the robustness of two S almonella LAMP assays in comparison with PCR and real‐time quantitative PCR for food applications. Methods and Results Performance of the assays was examined under abusive preparation conditions, running temperatures and p H , and with the addition of various inhibitors and food rinses. LAMP achieved robust detection under abusive assay preparation conditions (holding at 22 and 37°C for up to 30 min) and running temperatures (57–68°C). With a hot‐start DNA polymerase, PCR obtained comparable results under these temperature ranges. However, PCR performed markedly poorer under abusive pH . LAMP also showed greater tolerance to potential inhibitors than PCR . When food rinses including meat juice, chicken rinse, egg homogenate and produce homogenate were added at 20% of the reaction mix, PCR amplifications were completely inhibited, but LAMP reactions were not. Conclusions Our results demonstrated that LAMP is a robust alternative to PCR in S almonella detection for food applications. Significance and Impact of the Study This study filled important knowledge gaps regarding the robustness of S almonella LAMP assays. The findings will help bring S almonella LAMP assays closer to wider applications in food testing.