Establishment of PCR assays and genetic polymorphism analysis of genes encoding Clostridium perfringens β2 toxin from different sources.
10.3760/cma.j.cn112338-20220819-00718
- VernacularTitle:不同来源产气荚膜梭菌β
2毒素编码基因的PCR检测方法建立及遗传多态性分析
- Author:
Hao Ran ZHENG
1
;
Yuan Yuan WANG
2
;
Lu Lu BAI
2
;
Jia Xin ZHONG
2
;
Jin Xing LU
2
;
Yuan WU
2
;
Hui Ling DENG
3
Author Information
1. Shaanxi University of Traditional Chinese Medicine, Xi'an 712046, China National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
2. National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
3. Xi'an Central Hospital, Xi'an 710003, China.
- Publication Type:Journal Article
- MeSH:
Humans;
Clostridium perfringens/genetics*;
Clostridium Infections;
Bacterial Toxins/genetics*;
Phylogeny;
Polymerase Chain Reaction;
Polymorphism, Genetic
- From:
Chinese Journal of Epidemiology
2023;44(4):636-642
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To establish and optimize PCR methods for the gene encoding of Clostridium perfringens β2 toxin (cpb2) and atypical-cpb2 (aty-cpb2), analyze the epidemiological characteristics and genetic polymorphism of the cpb2 of Clostridium perfringens in 9 Chinese areas from 2016 to 2021. Methods: The cpb2 of 188 Clostridium perfringens strains were examined by PCR; the cpb2 sequences were acquired by whole-genome sequencing to analyze the genetic polymorphism. Using Mega 11 and the Makeblastdb tool, a phylogenetic tree, and cpb2-library based on 110 strains carrying the cpb2 were produced. Using the Blastn technique, a comparison was made to discover sequence similarity between consensus-cpb2 (con-cpb2) and aty-cpb2. Results: The specificity of PCR assay for the cpb2 and aty-cpb2 was verified. The PCR results for cpb2 amplification were highly consistent with the whole-genome sequencing approach (Kappa=0.946, P<0.001). A total of 107 strains from nine regions in China carried cpb2, 94 types A strains carried aty-cpb2, 6 types A strains carried con-cpb2, and 7 types F strains carried aty-cpb2. The nucleotide sequence similarity between the two coding genes was 68.97%-70.97%, and the similarity between the same coding genes was 98.00%-100.00%. Conclusions: In this study, a specific PCR method for cpb2 toxin was developed, and the previous PCR method for detecting aty-cpb2 was improved. aty-cpb2 is the primary gene encoding of β2 toxin. There is a significant nucleotide sequence variance between the various cpb2 genotypes.
