利用多重聚合酶链反应和多色毛细管电泳分析布氏菌多位点可变数目串联重复序列

发布时间：2019-06-20 02:10

【摘要】：目的传统的布鲁氏菌多点可变数目串联重复序列分析方法涉及16个位点(MLVA-16),包括单重聚合酶链反应和琼脂糖凝胶电泳,一直以来都作为标准的基因分型方法,用于布病的病原监测和流行病学调查。然而,目前这种传统方法工作量较大,实验室间结果不宜比较;尤其是对大规模菌株进行基因分型或面临突发疫情时,急需一种高通量且快速的方法。方法用多重PCR和多色毛细管电泳方法建立一种可靠的,高通量的且自动化程度较高的MLVA-16改良分型系统,用82株菌进行测试。结果这种改良的MLVA-16分型系统具有很高的准确性,且具有快速和高通量的特点。结论改良的MLVA-16分型系统可用于基层布病实验室,为布病的病原监测提供高效的工具。
[Abstract]:Objective the traditional method of multiple variable number tandem repeat analysis of Brucella involves 16 loci (MLVA-16), including single polymerase chain reaction (PCR) and agarose gel electrophoresis (agarose gel electrophoresis). It has been used as a standard genotyping method for pathogen surveillance and epidemiological investigation of brucellosis. However, at present, the workload of this traditional method is large, so it is not suitable to compare the results between laboratories, especially in the case of genotyping of large-scale strains or in the face of sudden outbreaks, a high-throughput and rapid method is urgently needed. Methods A reliable, high throughput and automatic MLVA-16 improved typing system was established by multiplex PCR and multicolor capillary electrophoresis. 82 strains of bacteria were tested. Results the improved MLVA-16 typing system had high accuracy, fast and high throughput. Conclusion the improved MLVA-16 typing system can be used in the laboratory of brucellosis at the grass-roots level, and provides an efficient tool for the pathogen monitoring of brucellosis.
【作者单位】：中国疾病预防控制中心传染病预防控制所传染病预防控制围家重点实验室感染性疾病协同诊治协同创新中心;中国疾病预防控制中心;
【基金】：国家自然科学基金(No.81271900) 国家卫生计生委公益性行业科研专项(No.201302006) 重大专项(No.2012ZX10004215)~~
【分类号】：R516.7

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