Novel motB as a potential predictive tool for identification of B-cereus, B-thuringiensis and differentiation from other Bacillus species by triplex real-time PCR

被引：10

作者：

Chelliah, Ramachandran ^{[1
]}

Wei, Shuai ^{[1
]}

Park, Byung-Jae ^{[1
]}

Kim, Se-Hun ^{[1
]}

Park, Dong-Suk ^{[2
]}

Kim, Soon Han ^{[3
]}

Hwan, Kim Seok ^{[3
]}

Oh, Deog-Hwan ^{[1
]}

机构：

[1] Kangwon Natl Univ, Dept Food Sci & Biotechnol, Chunchon 24341, South Korea

[2] Natl Acad Agr Sci, Dept Agr Biotechnol, Rural Dev Adm, Jeonju 54874, South Korea

[3] Natl Inst Food & Drug Safety Evaluat, Food Microbiol Div, 187,Osongsaengmyeong 2 Ro, Cheongju 28159, Chungcheongbuk, South Korea

来源：

MICROBIAL PATHOGENESIS | 2017年 / 111卷

关键词：

Bacillus; PCR (polymerase chain reaction); Identification; Genes; Rapid methods; CHARGED RESIDUES; FLAGELLAR; TAXONOMY; COMPLEX; FLIG;

D O I：

10.1016/j.micpath.2017.07.050

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

Quantitative triplex real-time PCR (qPCR) offers an alternative method for detection of bacterial contamination. It provides quantitation of the number of gene copies. In our study, we established a qPCR assay to detect and quantify the specificity towards Bacillus cereus and B. thuringiensis. The assay was designed to detect a 280 bp fragment of motB gene encoding the flagellar motor protein, specific for detection of B. cereus and B. thuringiensis, excluding other group species B. pseudomycoides, B. mycoides and B. weihenstephanensis. Specificity of the assay was confirmed with 111 strains belonging to Bacillus cereus group and performed against 58 B. cereus, 50 B. thuringiensis, 3 other Bacillus bacteria and 9 non Bacillus bacteria. Detection limit was determined for each assay. Direct analysis of samples revealed the specificity towards identification and characterization of B. cereus group cultured in nutrient media. Based on results, it was observed that motB showed 97% specificity towards B. cereus strains, 98% for B. thuringiensis but other B. cereus group showed less sensitivity (0%), thus, provides an efficient tool to identify B. cereus and B. thuringiensis. Further, environmental and food samples do not require band isolation, re-amplification or sequence identification. Thus, reducing the time and cost of analysis. (C) 2017 Elsevier Ltd. All rights reserved.

引用

页码：22 / 27

页数：6

共 26 条

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