Mining of novel target genes through pan-genome analysis for multiplex PCR differentiation of the major Listeria monocytogenes serotypes

被引：19

作者：

Li, Fan ^{[1
]}

Ye, Qinghua ^{[2
]}

Chen, Moutong ^{[2
]}

Zhou, Baoqing ^{[2
]}

Xiang, Xinran ^{[2
]}

Wang, Chufang ^{[2
]}

Shang, Yuting ^{[2
]}

Zhang, Jumei ^{[2
]}

Pang, Rui ^{[2
]}

Wang, Juan ^{[3
]}

Xue, Liang ^{[2
]}

Cai, Shuzhen ^{[2
]}

Ding, Yu ^{[4
]}

Wu, Qingping ^{[2
]}

机构：

[1] South China Univ Technol, Sch Biol & Biol Engn, Guangzhou, Peoples R China

[2] Guangdong Acad Sci, Guangdong Inst Microbiol, Guangdong Prov Key Lab Microbial Safety & Hlth, State Key Lab Appl Microbiol Southern China, Guangzhou, Peoples R China

[3] South China Agr Univ, Coll Food Sci, Guangzhou, Peoples R China

[4] Jinan Univ, Dept Food Sci & Technol, Guangzhou, Peoples R China

来源：

INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY | 2021年 / 339卷

基金：

中国国家自然科学基金;

关键词：

Comparative genomic method; Listeria monocytogenes; Serotyping; Multiplex PCR; Novel molecular target; LINEAGE; IDENTIFICATION; PRIMERS; 1/2A;

D O I：

10.1016/j.ijfoodmicro.2020.109026

中图分类号：

TS2 [食品工业];

学科分类号：

0832 ;

摘要：

The abundant information provided by the pan-genome analysis approach reveals the diversity among Listeria monocytogenes serotypes. The objective of this study was to mine novel target genes using pan-genome analysis for multiplex PCR detection and differentiation of the major L. monocytogenes serotypes present in food. Pan-genome analysis and PCR validation revealed a total of 10 specific targets: one for lineage I, two for serogroup I.1, one for serogroup I.2, two for lineage II, one for serogroup II.1, three for lineage III. Primers for the novel targets were highly specific in individual reactions. The detection limits were 10(3)-10(4) colony-forming units (CFU)/mL in pure bacterial cultures, meeting the requirements of molecular detection. Based on these novel targets, two new "lineage" multiplex PCR assays were developed to simultaneously distinguish between three lineages (I, II, and III) and five major serotypes (1/2a, 1/2b, 1/2c, 4b, and 4c) of L. monocytogenes. The detection limits of lineage I and lineage II&III mPCRs were 0.771 pg/mu L and 1.76 pg/mu L genomic DNA, respectively. The specificity of the mPCRs was robustly verified using other L. monocytogenes and non-L. monocytogenes serotypes. These results suggest that the two "lineage" multiplex PCRs based on novel targets offer a promising approach for accurate, sensitive, and rapid identification of L. monocytogenes serotypes.

引用

页数：9

共 32 条

[31]

Ward TJ, 2004, J BACTERIOL, V186, P4994, DOI [10.1128/JB.186.15.4994-5002.2004, 10.1128/jb.186.15.4994-5002.2004]

[32] A hybrid sub-lineage of Listeria monocytogenes comprising hypervirulent isolates [J].

Yin, Yuelan ;

Yao, Hao ;

Doijad, Swapnil ;

Kong, Suwei ;

Shen, Yang ;

Cai, Xuexue ;

Tan, Weijun ;

Wang, Yuting ;

Feng, Youwei ;

Ling, Zhiting ;

Wang, Guoliang ;

Hu, Yachen ;

Lian, Kai ;

Sun, Xinyu ;

Liu, Yuliang ;

Wang, Chuanbin ;

Jiao, Kuhua ;

Liu, Guoping ;

Song, Ruilong ;

Chen, Xiang ;

Pan, Zhiming ;

Loessner, Martin J. ;

Chakraborty, Trinad ;

Jiao, Xin'an .

NATURE COMMUNICATIONS, 2019, 10 (1)

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