返回 2008年第31卷第3期 目录

正交设计优化脑膜炎奈瑟氏菌PCR反应体系

杨 艳，叶运莉，苏红卫，甘仲霖，李爱玲，汪春梅，刘衡川1

（泸州医学院公共卫生系，四川泸州 646000；1四川大学公共卫生学院医学检验教研室）

摘 要 目的：建立脑膜炎奈瑟氏菌PCR检测的最佳反应体系，探讨影响检测的多个因素。方法：利用正交设计，对膜炎奈瑟氏菌PCR反应体系的4因素（Taq DNA聚合酶、引物、Mg2+、dNTP）在3个水平上进行优化实验，筛选出各反应因素的最佳水平。结果：正交实验结果表明，最佳反应体系为（50μl）：Taq DNA聚合酶2.0U，引物0.3μmol/L，Mg2+3.0mmol/L，dNTP100μmol/L。结论：所得体系反应稳定，正交法实验设计高效、快捷，科学性强，值得推广。

关键词 脑膜炎奈瑟氏菌；聚合酶链反应；正交设计；条件优化

OPTIMIZATION OF PCR REACTION SYSTEM FOR NEISSERIA MENINGITIDES USING ORTHOGONAL DESIGN

Yang Yan,et al

Department of Public Health，Luzhou Medical College

Abstract Objective: To establish a PCR optimal system for detecting Neisseria meningitides, and discuss multiple factors which affect this PCR assay.Methods: Orthogonal design was used to optimize the PCR amplification system for Neisseria meningitides in terms of four factors (Taq DNA polymerase, primer, Mg2+, dNTP) from three levels. Results: The suitable reaction conditions of this PCR were shown as follows:Taq DNA polymerase 2.0U,primer 0.3μmol/L,Mg2+ 3.0mmol/L,dNTP 100μmol/L for 50μl reaction system. Conclusion: The results showed that this method,which was invented to dissolve multi-factor-controlling problems, is simple, rapid,efficient,and promising.

Key words Neisseria meningitides; Polymerase chain reaction; Orthogonal design; Conditions optimization