先天致畸病原体感染多重PCR检测方法的建立

Multiplex PCR for detection of TORCH infection

目的 :建立先天致畸多种病原体 (TORCH)感染的多重PCR诊断方法。方法 :选择弓形虫、巨细胞病毒、Ⅰ型和Ⅱ型单纯疱疹病毒特异性基因片段 ,按照多重PCR引物设计的特殊要求 ,各设计一对引物 ,以四种病原体标准株为模板进行扩增。找出各病原体最佳扩增条件 ,再进行综合分析 ,最终找到一个理想的多重PCR扩增条件。结果 :各病原体扩增片段及多重PCR扩增条带均清晰、均一 ,产量较高 ,无非特异扩增 ,片段长度与理论值一致 ;DNA测序证实 ,各扩增产物就是各病原体特异性基因片段。结论 :本研究成功地建立TORCH感染敏感、特异、快捷的多重PCR实验诊断方法 ,对优生优育工作的开展具有十分重要的意义。

Objective:To establish a multiple polymerase chain reaction(multiplex PCR) technique for detection of TORCH infection.Methods:The specific gene fragments of toxoplasma gondii,cytomegalovirus and herpes simplex virus type 1,2 were chosen and four pairs of primers were designed.After every fragment was amplified from template DNA of standard strains,the appropriate condition of multiplex PCR was determined.Results:The bands of four products were seen in agarose gel electrophoresis clear and unique,without nonspecific amplified fragment,the base pairs and DNA sequences of these four bands were in coincidence with lengthes and sequences which had been predicted.Conclusions:A specific,sensitive and fast multiplex PCR method for detection of TORCH infection had been established. [