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目的为患者提供一种新型的多重(19种)腹泻病原体的检测方案,用于临床快速诊断。方法利用NCBI数据库设计19种腹泻病原体及内参的特异性引物;建立多重逆转录-聚合酶链式反应(mRT-PCR)体系;用GeXP遗传分析系统对PCR产物进行毛细管电泳分离,从而对19种腹泻病原体进行鉴别。结果 100例腹泻样本中阳性样本为85例,阴性样本为15例,共出现单病原体感染32例,混合感染53例,与测序结果一致。19种病原体特征峰均有出现,且各个峰之间分离良好,未见交叉现象。结论利用GeXP分析平台联合mRT-PCR技术同时检测19种腹泻病原体的方法,具有高通量、灵敏度高、特异性强且检验速度快等优点,此方法有可能满足临床医生对腹泻病原体检测的需求,从而指导临床治疗。
Objective To provide patients with a new multiple (19) diarrhea pathogen detection program for rapid clinical diagnosis. Methods 19 kinds of specific primers for pathogen and internal reference of diarrhea were designed by using NCBI database. Multiple reverse transcription-polymerase chain reaction (mRT-PCR) system was established. The PCR products were separated by capillary electrophoresis using GeXP genetic analysis system, Diarrhea pathogens were identified. Results The positive samples of 100 cases of diarrhea were 85 cases and the negative samples were 15 cases. There were 32 cases of single pathogen infection and 53 cases of mixed infection, which were consistent with the sequencing results. The characteristic peaks of 19 kinds of pathogens all appeared, and the peaks were well separated with no crossover phenomenon. Conclusion The method of simultaneous detection of 19 diarrhea pathogens by the GeXP analysis platform combined with mRT-PCR technology has the advantages of high throughput, high sensitivity, strong specificity and fast test speed, which may meet clinicians’ needs for the detection of diarrhea pathogens , To guide clinical treatment.