型特异性多重PCR快速检测创伤组织中的产气荚膜梭菌

目的 为快速检测创伤组织中的产气英膜梭菌建立型特异性多重PCR方法.方法 建立菌落和含产气荚膜梭菌创伤标本中的染色体和质粒DNA简便、快速纯化[月桂基硫酸三乙胺醇(TLS)法]和型特异性多重PCR方法;对各型产气荚膜梭菌、部分近缘梭菌标准株和开放性创伤气性坏疽标本进行PCR检测,其结果与培养结果进行比较.结果 该方法可检测各型产气荚膜梭菌,并可排除近缘梭菌干扰;A型株检测灵敏度达到7.5×10~3/ml;可在5 h内获取结果;标本PCR检测与培养结果相符.结论 新建立的培养和创伤组织标本中产气荚膜梭菌染色体与质粒DNA纯化方法简便、快速;型特异性多重PCR具有良好的特异性、较高的灵敏度,短时间内获得结果,可用于临床实验室.

A Rapid Method by Type-specific Multiplex PCR for Clostridium perfringens on Surface of Traumatic Tissues

OBJECTIVE To establish a rapid detecting method for Clostridium perfringens on traumatic tissues by type-specific multiplex PCR. METHODS Established a simple and rapid method (TLS method) for purifying the DNA of genomes and plasmids in standard strains of C. perfringens wild strains on surface of open traumatic tissues and detected DNAs by type-specific multiplex PCR. RESULTS All types of C. perfringens could be detected by type-specific multiplex PCR. The sensitivity by PCR for type A of C. perfringens arrived at 7.5× 10~3/ml and a whole run could finish within 5 h; the results by PCR entirely corresponded with those by cultureing. CONCLUSIONS New methods for purifying DNA of genomes and plasmids of C. perfringens are simple and rapid; there are high specificity and sensitivity for detecting DNA by multiplex PCR within short time, which can be practiced in clinical laboratory.