利用Luminex xMap悬浮阵列对炭疽芽孢杆菌进行多重基因分型。

Multiplexed genotyping of Bacillus anthracis by Luminex xMap suspension array.

作者信息

Thierry Simon, Derzelle Sylviane

机构信息

Anses, Animal Health Laboratory, Bacterial Zoonosis Unit, University Paris-Est, 23 avenue du Général De Gaulle, 94706, Maisons-Alfort, France,

出版信息

Methods Mol Biol. 2015;1247:401-12. doi: 10.1007/978-1-4939-2004-4_29.

DOI:10.1007/978-1-4939-2004-4_29

PMID:25399112

Abstract

The Luminex® xTAG technology is a medium to high throughput, open methodology able to test many single nucleotide polymorphisms (SNPs) in a single reaction and a minimum time. Multiplex SNPs interrogation are conducted on the Luminex xMAP system, which uses lasers to read universal tag, color-coded microspheres that attach to specific nucleic acid sequences. The present method describes a Multiplex Oligonucleotide Ligation-PCR procedure (MOL-PCR) for the simultaneous interrogation of 13 phylogenetically informative SNPs within the genome of Bacillus anthracis. The reported 13-plex assay enables efficient B. anthracis genotyping into major sublineages and groups. While cost-effective compared to other monoplex methods, the present MOL-PCR method also offers a high degree of flexibility and scalability. It can easily accommodate newly identified SNPs to increase resolving power to the canSNP typing of B. anthracis.

摘要

Luminex® xTAG技术是一种中高通量的开放式方法，能够在单个反应中以最短时间检测多个单核苷酸多态性（SNP）。多重SNP检测在Luminex xMAP系统上进行，该系统使用激光读取与特定核酸序列结合的通用标记、彩色编码微球。本方法描述了一种多重寡核苷酸连接-PCR程序（MOL-PCR），用于同时检测炭疽芽孢杆菌基因组内13个系统发育信息丰富的SNP。所报道的13重检测法能够将炭疽芽孢杆菌高效基因分型为主要亚谱系和组。与其他单重方法相比，本MOL-PCR方法具有成本效益，还具有高度的灵活性和可扩展性。它可以轻松容纳新鉴定的SNP，以提高炭疽芽孢杆菌canSNP分型的分辨能力。