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Brucella melitensis and Brucella abortus genotyping via real-time PCR targeting 21 variable genome loci
Journal of Microbiological Methods ( IF 1.7 ) Pub Date : 2020-12-14 , DOI: 10.1016/j.mimet.2020.106125
Selçuk Kılıç 1 , Bekir Çelebi 2 , Meral Turan 3
Affiliation  

Brucella melitensis and Brucella abortus account for almost all cases of brucellosis in Turkish population. We developed a fourplex quantitative real-time PCR (qPCR) assay for the electrophoresis-free, rapid and cost-effective differentiation of B. abortus and B. melitensis from the other Brucella spp. The 4-plex species differentiation assay was combined with a qPCR assay targeting 17 different single nucleotide polymorphism (SNP) loci in Brucella genomes. This combination resulted in a 21 Variable Genome Loci (21-VGL) qPCR assay for high resolution genotyping of B. abortus and B. melitensis. A total of 486 Brucella was analyzed using the qPCR assay to create a 21-VGL profile database. The database contained the profiles of 55 B. abortus, 352 B. melitensis, 3 B. ceti, 6 B. neotomae, 7 B. ovis, 6 B. pinnipedialis, 44 B. suis and 13 B. canis strains. The 21-VGL Brucella genotyping clearly distinguished B. abortus, B. melitensis, B. neotomae and B. ovis. The 21-VGL approach could not distinguish B. pinnipedialis from B. ceti and some B. suis genotypes from B. canis. The results revealed that more than 99% of the Brucella isolates in Turkey were B. melitensis and 21-VGL genotyping can be reduced to 8-VGL B. melitensis genotyping without any loss of genotyping resolution. To our knowledge, we introduced the fastest and the lowest-cost B. abortus and B. melitensis genotyping and species differentiation methodology in the literature.



中文翻译:

通过针对 21 个可变基因组位点的实时 PCR 对羊布鲁氏菌和流产布鲁氏菌进行基因分型

布鲁氏菌流产布鲁氏几乎是土耳其人群中所有布鲁氏菌病的原因。我们开发了一种四重定量实时 PCR (qPCR) 检测方法,用于无需电泳、快速且经济高效地将B. abortusB. melitensis与其他布鲁氏菌属进行区分。4 重物种分化测定与针对布鲁氏菌基因组中17 个不同单核苷酸多态性 (SNP) 位点的 qPCR 测定相结合。这种组合产生了 21 个可变基因组位点 (21-VGL) qPCR 检测,用于对流产 B. abortusB. melitensis进行高分辨率基因分型。共486个布鲁氏菌使用 qPCR 测定法分析以创建 21-VGL 配置文件数据库。该数据库包含 55 种B. abortus、352 B. melitensis、3 B.ceti、6 B. neotomae、7 B. ovis、6 B. pinnipedialis、44 B. suis和 13 B. canis菌株的概况。这位21 VGL布鲁氏菌基因分型明确区分B.流产B.羊种B. neotomaeB.绵羊。这位21 VGL的做法根本无法辨别B. pinnipedialisB. CETI有的B.猪B. canis 的基因型。结果表明,土耳其99% 以上的布鲁氏菌分离株是B. melitensis,21-VGL 基因分型可以减少到 8-VGL B. melitensis基因分型,而不会降低基因分型分辨率。据我们所知,我们介绍了文献中最快、成本最低的B. abortusB. melitensis基因分型和物种分化方法。

更新日期:2020-12-25
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