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.

布鲁氏菌和流产布鲁氏菌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. abortus和B. 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. abortus和B. 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. neotomae和B.绵羊。这位21 VGL的做法根本无法辨别B. pinnipedialis从B. 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. abortus和B. melitensis基因分型和物种分化方法。