利用Lambda_Ba03噬菌体基因实时荧光定量PCR检测炭疽芽孢杆菌

Q3 Medicine
A. Nizkorodova, E. R. Mal’tseva, Z. Berdygulova, D. A. Naizabaeva, S. Kuatbekova, A. Zhigailov, N. Abdolla, A. Mashzhan, I. A. Akhmetollaev, Y. Skiba, S. Mamadaliev
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引用次数: 0

摘要

本研究的目的是建立一套基于lambda_Ba03噬菌体基因的实时荧光PCR检测炭疽芽孢杆菌两个染色体靶点的引物和荧光探针。材料和方法。BLAST分析发现,在lambdaBa03原噬菌体区域存在BA_5358 (AE016879.1: 4852332. 4853642)和BA_5361 (AE016879.1: 4855298. 4856278)两个靶基因。设计的引物和荧光水解TaqMan探针用于同时检测两种基因的炭疽芽胞杆菌染色体DNA,并进行qPCR检测其敏感性和特异性。结果和讨论。对密切相关的细菌(蜡样芽孢杆菌、苏云金芽孢杆菌、枯草芽孢杆菌、克劳氏芽孢杆菌)的染色体DNA样本进行的研究表明,所开发的引物/探针具有100%的特异性。在m55-VNIIVViM疫苗株DNA样本和炭疽芽孢杆菌档案DNA样本上,设计的多重检测试剂盒的灵敏度达到100 fg,每次反应的灵敏度上限为17个基因组。开发的多重试剂盒可作为研究炭疽的实验室的单独工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Real-Time PCR Detection of Bacillus anthracis by Lambda_Ba03 Prophage Genes
The aim of the study was to develop a set of primers and fluorescent probes for the detection of two chromosomal targets of Bacillus anthracis using real-time PCR based on the lambda_Ba03 prophage genes.Materials and methods. BLAST analysis of B. anthracis chromosomal DNA identified two target genes in the region of lambdaBa03 prophage, BA_5358 (AE016879.1: 4852332..4853642) and BA_5361 (AE016879.1: 4855298..4856278). The designed primers and fluorescent hydrolysable TaqMan probes for simultaneous detection of B. anthracis chromosomal DNA by two stated genes were tested in qPCR for sensitivity and specificity.Results and discussion. Studies performed on chromosomal DNA samples of closely related bacteria (B. cereus, B. thuringiensis, B. subtilis, B. clausii) have shown 100 % specificity of the developed sets of primers/probes. The sensitivity of the devised multiplex kit, tested on DNA samples of the m55-VNIIVViM vaccine strain and archival DNA samples of B. anthracis, reached 100 fg of bacterial DNA, which sets the limit of sensitivity at 17 genomes per reaction. The developed multiplex kit can be used as a separate tool for research laboratories studying anthrax.
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来源期刊
Problemy Osobo Opasnykh Infektsii
Problemy Osobo Opasnykh Infektsii Medicine-Infectious Diseases
CiteScore
1.90
自引率
0.00%
发文量
79
审稿时长
12 weeks
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