Premium
Development of a novel dual priming oligonucleotide system‐based PCR assay for specific detection of Salmonella from food samples
Author(s) -
Li DanDan,
Hao ChunBo,
Liu ZhongMei,
Wang SuiJia,
Wang Yu,
Chao Zhe,
Gao ShenYang,
Chen Si
Publication year - 2020
Publication title -
journal of food safety
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.427
H-Index - 43
eISSN - 1745-4565
pISSN - 0149-6085
DOI - 10.1111/jfs.12789
Subject(s) - salmonella , oligonucleotide , polymerase chain reaction , detection limit , priming (agriculture) , microbiology and biotechnology , biology , chemistry , chromatography , bacteria , gene , biochemistry , genetics , germination , botany
In this study, a novel dual priming oligonucleotide (DPO) system‐based polymerase chain reaction (PCR; DPO system‐based PCR) assay, which detected the fimY gene of Salmonella , was developed for the fast food testing. The DPO system‐based PCR assay allowed a wide range of annealing temperatures at 48–68°C to efficiently amplify fimY gene with an analytical detection limit of 1.2 × 10 2 CFU/ml for Salmonella in pure cultures and artificially contaminated food matrix. Significantly, the presence of a bubble‐like polydeoxyinosine (polyI) linker in the DPO system brought an unparalleled high specificity in the identification of target bacteria, and consequently, the false positives and mismatches of PCR process can be eliminated in priming. Applying the DPO system‐based PCR assay to 285 collected food samples revealed that 29 samples were positive in this assay, in accordance with the results of conventional culture‐based method, indicating a potential diagnostic capability. The high specificity of the DPO system‐based PCR indicates its great potential to be a quick, reliable and practical method for the detection of Salmonella in foods.