布鲁氏菌多位点可变数目串联重复序列分析方法的改良  

Optimization of multi-locus variable number tandem repeat sequence analysis method for Brucella

作　　者：谭勤琴 王月[2] 刘英[2] 杨幸贵 营夏 胡勇[1] 李世军[1,2] TAN Qin-qin;WANG Yue;LIU Ying;YANG Xing-gui;YING Xia;HU Yong;LI Shi-jun(不详;School of Public Health,the Key Laboratory of Environmental Pollution Monitoring and Disease Control,Ministry of Education,Guizhou Medical University,Guiyang,Guizhou 550025,China)

机构地区：[1]贵州医科大学公共卫生与健康学院,环境污染与疾病监控教育部重点实验室,贵州贵阳550025 [2]贵州省疾病预防控制中心,细菌性检验科

出　　处：《现代预防医学》2022年第23期4372-4378,共7页Modern Preventive Medicine

基　　金：国家自然科学基金项目(82273758);贵州省传染病人才培养基地建设项目子项目(RCJD2102)。

摘　　要：目的为简化传统布鲁氏菌多位点可变数目串联重复序列(Multiple locus variable-number tandem repeat analysis,MLVA)分析方法,运用多重聚合酶链反应结合多色毛细管电泳技术进行方法改良。方法以贵州省83株布鲁氏菌地方株及3株疫苗株(M5、A19、S2)为配对试验研究对象,同时运用两种检测技术,分为对照组(传统单重PCR结合毛细管电泳技术,8个反应体系)及试验组(多重PCR结合多色毛细管电泳技术,3个反应体系)对布鲁氏菌MLVA panel 2模块的8个位点进行检测,将试验组同对照组间的检测数据及试验组同预期值间数据分别用统计学方法进行配对分析。结果试验组同对照组间的检测数据及试验组同预期值间的数据经配对检验,P值分别为0.072、0.192,差异均不具有统计学意义(P>0.05)。该改良方法通过缩减反应体系,简化操作步骤,适用于基层实验室,且具有结果可靠、操作简单及高通量的特点。结论针对布鲁氏菌panel 2模块位点设计的多重聚合酶链反应结合多色毛细管电泳技术能更高效、更便捷用于布病溯源得分析。Objective To simplify the traditional Brucella multi-locus variable number tandem repeat sequence analysis method,which is modified using a multiplex polymerase chain reaction combined with multicolor capillary electrophoresis.Methods Three vaccine strains(M5,A19,and S2)and the 83 Brucella strains,preserved by the Guizhou Provincial,were used for the study.Two assay techniques were used simultaneously,a control group(single PCR combined with capillary electrophoresis)and a test group(multiplex PCR combined with multicolor capillary electrophoresis)for the determination of 8 loci of the Brucella MLVA panel 2 modules.The test results between the test group and the control group and between the test group and the standard reference value were analyzed by statistical methods.Results The test data,between the test group and the control group and between the test group and the standard reference value,were respectively tested by paired Wilcoxon signed-rank sum test with P values of 0.072 and 0.192 respectively,the differences were not statistically significant(P>0.05).The improved method reduced the reaction system and simplified the operation steps.It is suitable for grass-roots laboratories,and has the characteristics of reliable results,simple operation,and high throughput.Conclusion Multiplex polymerase chain reaction combined with multicolor capillary electrophoresis for Brucella panel 2 module loci can be more efficient and easier to use for Brucella traceability analysis.

关 键 词：布鲁氏菌 MLVA Wilcoxon符号秩和检验 

分 类 号：R387.5[医药卫生—医学寄生虫学]