布鲁氏菌PFGE分子分型的条件优化

MOLECULAR TYPING OF BRUCELLA BY PULSED-FIELD GEL ELECTROPHORESIS(PFGE)

[目的]将PFGE方法应用于布鲁氏菌分型研究,探索建立布鲁氏菌PFGE分型的最优化方法。[方法]在用PFGE方法进行布鲁氏菌基因分型的研究中,针对不同内切酶、酶切的浓度和时间、脉冲时间等影响PFGE分型结果的因素进行优化,利用优化后的条件对布鲁氏菌19株标准菌株进行分型研究。[结果]在布鲁氏菌的PFGE分型研究中,选择XbaI为内切酶、酶切浓度为96u/200μl、酶切时间为4h、脉冲时间为2～25s为PFGE电泳的优化条件,在分析的19株布鲁氏菌标准菌株中,6种布菌的PFGE图谱各不相同,但PFGE不能将种内不同生物型布菌完全区分开来。[结论]PFGE方法可用于布鲁氏菌的基因分型的研究,是对传统分型方法一种较好的补充。

[Objective] To establish and optimize a method of molecular typing of Brucella by pulsed-field gel electrophoresis （PFGE）. [Methods] PFGE was used in the genetic typing study of Brucella to optimizing the factors influencing the typing of PFGE including different restriction endonuclease, different reaction concentration and reaction time. With these optimized factors, typing research was conducted to the 19 reference strains of Brucella. [ Results] Restriction endonucleases XbaI was selected in this study with the optimized condition tO be reaction concentration of 96u/200μl, reaction time of 4 hours and pulse time of 2s-25s. By this technique, six electrophoretypes were distinguished in six reference strains of the different species, but the PFGE could not distinguish the Brucella within a species. [Conclusion] PFGE could be used to classify and identify Brucella, which is a good supplementary for the tradition typing method.