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。
The Multi Locus Variable Number Tandem Repeat Analysis 16 loci panel (MLVA-16), involving singleplex PCRs and agarose gel electrophoresis, is the standard genotyping method for Brucella spp. used also for the Brucella interna- tional online database. To find a rapid and through-put genotyping method, modified MLVA-16 can be used. We described an alternative, reliable, high-throughput MLVA-16 protocol using multiplex PCRs and multicolor capillary electrophoresis. Fur- thermore, 82 Brucella strains were tested. Results showed that the modified MLVA-16 has proved to be accurate, rapid and throughput. It's indicated that the modified MLVA-16 is highly discriminatory and epidemiological concordance and is easy for brucellosis surveillance in province-level laboratory.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2015年第4期303-310,共8页
Chinese Journal of Zoonoses
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)~~