环介导等温扩增技术在免疫功能低下患儿中快速检测耶氏肺孢子菌感染的研究

Rapid detection of pneumocystis carinii infection in immunocompromised patients by loop mediated isothermal amplification technique

目的探讨环介导等温扩增(LAMP)技术在免疫功能低下患儿耶氏肺孢子菌(PC)快速病原检测中的应用价值。方法收集2020年5月至2021年5月在上海交通大学医学院附属上海儿童医学中心治疗的免疫功能低下、根据临床表现高度怀疑耶氏肺孢子菌肺炎(PCP)患儿的肺泡灌洗液或深部痰液, 应用聚合酶链反应(PCR)及LAMP方法检测PC。根据PC基因保守区域合成LAMP引物, 并对LAMP反应体系及反应条件进行优化, 评价其敏感性与特异性, 将其与PCR检测病原体结果进行比较。结果本研究所建立的PC LAMP检测技术特异性和敏感性均较高, 1h即可出检测结果。LAMP技术与PCR检测临床高度怀疑PCP患儿12份临床标本, 阳性10例, 阴性2例, 二者符合率100%。结论 LAMP技术可快速高效检测PC, 较普通PCR检测效率高, 可为临床快速诊断PCP提供良好的技术手段。

Objective To explore the value of detecting pneumocystis carini(PC)rapidly in immunocompromised patients by loop mediated isothermal amplification(LAMP).Methods Respiratory tract specimens of immunocompromised children suspected of pneumocystis carinii pneumonia(PCP)at Shanghai Children′s Medical Center,School of Medicine,Shanghai Jiao Tong University were collected from May 2020 to May 2021.PCR and LAMP methods were used to detect PC.Firstly,LAMP primers of PC were synthetized according to the conserved region of PC gene,and the LAMP reaction system and reaction conditions were optimized to evaluate the sensitivity and specificity.Then,the results of pathogens were compared with those of PCR detection.Results The established LAMP detection technology for PC had high specificity and super sensitivity.The detection results could be obtained within 1 hour.In 12 clinical samples,10 cases were positive and 2 cases were negative,the coincidence rate of LAMP and PCR technique was 100%.Conclusion LAMP can detect PC more rapidly and sensitively than PCR,and it can provide a good support for clinical rapid diagnosis of PCP.