Moradkasani Safoura, Jazi Faramarz M, Sadeghifard Nourkhoda, Kouhsari Ebrahim, Kalani Behrooz S, Pakzad Iraj
Clin Lab. 2020 Oct 1;66(10). doi: 10.7754/Clin.Lab.2020.200119.
Brucellosis is considered a main health concern in humans and animals. Neither familiar molecular methods nor the classical biotyping techniques are acceptable for subtyping Brucella spp. Loci containing variable number tandem repeats (VNTRs) have recently demonstrated their practicality in typing isolates from human and animal origin despite the excessive genetic homogeneity in the genus Brucella.
The genotypic characteristics of sixty-six Brucella melitensis and thirty-four Brucella abortus isolates from veterinary samples and human brucellosis cases in Iran during 2014 - 2018. They were analyzed using multiple-locus variable-number tandem-repeat analysis (MLVA) which consisted of sixteen primer pairs and designed and classified as belonging to one of the three panels: panel 1 (MLVA-8: eight loci including Bruce06, Bruce08, Bruce11, Bruce12, Bruce42, Bruce43, Bruce45, and Bruce55), panel 2A (three loci including Bruce18, Bruce19, and Bruce21), and panel 2B (five loci including Bruce04, Bruce07, Bruce09, Bruce16, and Bruce30); MLVA-11 (panels 1 and 2A), and MLVA-16 (panels 1, 2A, and 2B) using BioNumerics software (Version 7.6).
Using panel 1, 2A, and 2B (MLVA-16), 59 genotypes with a genetic similarity coefficient ranging from 91 to 100% were obtained from the 100 Brucella spp. isolates. For all isolates, only genotype 36 and genotype 26 were obtained using panels 1 and 2A, respectively. The B. abortus isolates showed variations at 9 different genotypes, while B. melitensis isolates have been dispersed in 50 different genotypes. Bruce16 and Bruce4 showed the highest discriminatory power.
The MLVA-16 assay appeared to be a useful and important molecular genotyping tool that is capable of proving epidemiological linkages in outbreak and trace-back investigations and is helpful in improving the effectiveness of brucellosis control programs.
布鲁氏菌病被认为是人和动物的主要健康问题。无论是常见的分子方法还是经典的生物分型技术都不适用于布鲁氏菌属的亚型分类。尽管布鲁氏菌属存在过度的基因同质性，但含有可变数目串联重复序列（VNTRs）的基因座最近已证明其在对来自人和动物源的分离株进行分型方面的实用性。
对2014年至2018年期间从伊朗的兽医样本和人类布鲁氏菌病病例中分离出的66株羊种布鲁氏菌和34株牛种布鲁氏菌进行基因型特征分析。使用多位点可变数目串联重复序列分析（MLVA）对它们进行分析，该分析由16对引物组成，并设计并归类为属于三个面板之一：面板1（MLVA - 8：8个基因座，包括Bruce06、Bruce08、Bruce11、Bruce12、Bruce42、Bruce43、Bruce45和Bruce55），面板2A（3个基因座，包括Bruce18、Bruce19和Bruce21），以及面板2B（5个基因座，包括Bruce04、Bruce07、Bruce09、Bruce16和Bruce30）；使用BioNumerics软件（版本7.6）进行MLVA - 11（面板1和2A）和MLVA - 16（面板1、2A和2B）分析。
使用面板1、2A和2B（MLVA - 16），从100株布鲁氏菌属分离株中获得了59种基因型，遗传相似系数范围为91%至100%。对于所有分离株，分别使用面板1和2A仅获得了基因型36和基因型26。牛种布鲁氏菌分离株显示出9种不同基因型的变异，而羊种布鲁氏菌分离株分布在50种不同基因型中。Bruce16和Bruce4显示出最高的鉴别力。
MLVA - 16检测似乎是一种有用且重要的分子基因分型工具，能够在疫情爆发和追溯调查中证明流行病学联系，并有助于提高布鲁氏菌病控制项目的有效性。
