文献类型： 外文期刊

作者： Liu, Junyan ¹ ; Xiang, Zhufang ⁴ ; Huang, Tengyi ³ ; Xu, Zhenbo ³ ; Liu, Gongliang ¹ ; Qu, Chunyun ¹ ; Soteyome, Thanapop ⁵ ; Luo, Yuting ⁶ ; Ma, Qin ⁷ ; Yuan, Lei ⁸ ; Hong, Wei ⁹ ; Li, Xuejie ⁶ ; Ye, Yanrui ¹¹ ;

作者机构： 1.Zhongkai Univ Agr & Engn, Acad Contemporary Agr Engn Innovat, Coll Light Ind & Food Sci, Guangdong Prov Key Lab Lingnan Specialty Food Sci, Guangzhou 510225, Peoples R China

2.Minist Agr, Key Lab Green Proc & Intelligent Mfg Lingnan Speci, Guangzhou 510225, Peoples R China

3.Shantou Univ, Affiliated Hosp 2, Med Coll, Dept Lab Med, Shantou, Guangdong, Peoples R China

4.Guangzhou Zhaopin Feed Co Ltd, Guangzhou 511400, Guangdong, Peoples R China

5.Rajamangala Univ Technol Phra Nakhon, Home Econ Technol, Bangkok, Thailand

6.South China Univ Technol, Engn Res Ctr Starch & Vegetable Prot Proc, Sch Food Sci & Engn, Minist Educ,Guangdong Prov Key Lab Green Proc Nat, Guangzhou 510640, Peoples R China

7.Guangdong Acad Agr Sci, Sericultural & Agrifood Res Inst, Key Lab Funct Foods, Minist Agr,Guangdong Key Lab Agr Prod Proc, Guangzhou 510610, Peoples R China

8.Yangzhou Univ, Sch Food Sci & Engn, Yangzhou 225127, Jiangsu, Peoples R China

9.Guangzhou Med Univ, GMU GIBH Joint Sch Life Sci, Guangzhou, Guangdong, Peoples R China

10.Res Inst Food Nutr & Human Hlth, Guangzhou, Peoples R China

11.South China Univ Technol, Sch Biol & Biol Engn, Guangzhou, Peoples R China

关键词： Vibrio parahaemolyticus; Food safety; Polymerase spiral reaction (PSR); Rapid detection; VBNC cells

期刊名称：LWT-FOOD SCIENCE AND TECHNOLOGY （ 影响因子：6.0； 五年影响因子：6.0 ）

ISSN： 0023-6438

年卷期： 2024 年 198 卷

页码：

收录情况： SCI

摘要： Vibrio parahaemolyticus is a predominate cause of microbiological food safety. This study aimed to develop a polymerase spiral reaction (PSR) based assay for rapid and visual detection on V. parahaemolyticus . Firstly, the specificity and conservativity of species-specific target tlh gene were determined. Secondly, targeting tlh and virulence factors tdh and trh , PSR assay was developed and optimized using constructed positive plasmid controls and standard strain. Visualized result determination using fluorescent dye was applied to avoid false positive issues. Thirdly, 44 strains were used for specificity evaluation. Fourthly, pure and mixed species artificial contamination systems were included to test its applicability. In addition, a propidium monoazide (PMA)-PSR assay was set up to determine viable but non-culturable (VBNC) cells. Target tlh gene acquired high specificity and PSR assay was optimized to carry out at 65 degrees C for 1 h. Detection limit of PSR assay in purified genomic DNA, plasmid and artificially contaminated food samples were 5.31-53.1 pg/mu L, 100 copies, and 10(3) CFU/mL, respectively, demonstrating the sensitivity as 100-fold higher than regular PCR. With PMA-PSR assay, VBNC cells were accurately determined without the disruption of dead cells and food compositions. Thus, PSR is applicable in detection of V. parahaemolyticus viable cells and virulence factors.