宏基因组学二代测序在艾滋病合并肺部感染患者中的临床应用

Clinical application of metagenomic next-generation sequencing in acquired immunodeficiency syndrome patients with pulmonary infections

目的了解本地区艾滋病患者合并肺部机会性感染的病原谱, 并评价宏基因组学二代测序(mNGS)在艾滋病合并肺部感染患者中的临床应用。方法以2021年1月至12月在武汉大学中南医院住院的艾滋病合并肺部感染患者为研究对象, 对其支气管肺泡灌洗液(BALF)进行mNGS和常规病原学检测。常规病原学检测方法包括涂片、培养、聚合酶链反应(PCR)和免疫层析胶体金法。统计学方法采用Fisher确切概率法。结果共纳入69例患者, 其mNGS检测结果均为阳性, 其中真菌和病毒各检测出53例次(76.8%), 细菌[不包括结核分枝杆菌(MTB)和非结核分枝杆菌(NTM)]40例次(58.0%), MTB 6例次, NTM 11例次, 其他7例次;89.9%(62/69)的患者为2种或以上病原体的混合感染。BALF的常规病原学检测中, 79.7%(55/69)的患者检出病原体阳性, 包括耶氏肺孢子菌聚合酶链反应(PCR)检测阳性42例次, BALF培养阳性16例次, MTB PCR检测阳性9例次, 隐球菌抗原阳性5例次。mNGS总检出率为100.0%(69/69), 高于常规病原学检测的79.7%(55/69), 差异有统计学意义(Fisher确切概率法, P<0.001)。mNGS检测特异度为88.4%。结合临床及2种检测方法, 前5位的病原体分别为耶氏肺孢子菌[62.3%(43/69)]、念珠菌[29.0%(20/69)]、MTB[20.3%(14/69)]、NTM和马尔尼菲篮状菌[各15.9%(11/69)]。76.8%(53/69)的患者合并病毒感染。结论本地区艾滋病患者合并肺部感染的病因以混合感染为主, 耶氏肺孢子菌为最常见的病原体。mNGS可提高艾滋病合并肺部感染患者的病原体检出率。

Objective To investigate the pathogen spectrum of acquired immunodeficiency syndrome(AIDS)patients with pulmonary opportunistic infections in the local area,and to evaluate the clinical application of metagenomic next-generation sequencing(mNGS)in these patients.Methods From January to December 2021,AIDS patients with pulmonary infections admitted to Zhongnan Hospital of Wuhan University were enrolled.Their bronchoalveolar lavage fluid(BALF)was subjected to mNGS and coventional pathogen detection.Routine pathogen detection methods included smear,culture,polymerase chain reaction(PCR),and immunochromatographic colloidal gold.Fisher′s exact probability method was used for statistical analysis.Results A total of 69 patients were included,and all of them were tested positive for mNGS.Among them,53 cases(76.8%)were positive for fungi and viruses,40 cases(58.0%)were positive for bacteria(excluding Mycobacterium tuberculosis(MTB)and nontuberculous mycobacteria(NTM)),six cases were positive for MTB,11 cases were positive for NTM,and seven cases were positive for other pathogens.Mixed infections with two or more pathogens were found in 89.9%(62/69)of the patients.Among the conventional pathogen detections of BALF,79.7%(55/69)of the patients were positive for pathogens,including 42 cases positive for Pneumocystis jirovecii PCR,16 cases positive for BALF culture,nine cases positive for MTB PCR,and five cases positive for Cryptococcus antigen.The total detection rate of mNGS was 100.0%(69/69),which was higher than that of the conventional pathogen detection rate of 79.7%(55/69),and the difference was statistically significant(Fisher′s exact probability method,P<0.001).The specificity of mNGS detection was 88.4%.Combining clinical and two detection methods,the top five pathogens were Pneumocystis jirovecii(62.3%(43/69)),Candida(29.0%(20/69)),MTB(20.3%(14/69)),NTM and Talaromyces marneffei(15.9%(11/69),each).Fifty-three patients(76.8%)had co-infection with virus.Conclusions The main cause of pulmonary infection in AIDS patients in this area is mixed infection,and Pneumocystis jirovecii is the most common pathogen.mNGS could significantly improve the pathogen detection rate in AIDS patients with pulmonary infections.