病原真菌real-time PCR检测对非中性粒细胞缺乏的老年患者肺部真菌病的诊断价值  

The real-time PCR of fungal pathogens in bronchoalveolar lavage fluid for the diagnosis of pulmonary fungal diseases in non-neutropenic elderly patients

作　　者：蒙星烨 刘晓[1] 万喆[1] 陈伟[1] 阙呈立[2] 林连君[3] 余进[1] 宋营改[1] 李若瑜[1] MENG Xingye;LIU Xiao;WAN Zhe;CHEN Wei;QUE Chengli;LIN Lianjun;YU Jin;SONG Yinggai;LI Ruoyu(Departmentof Dermatology and Venerology,Peking University First Hospital,Research Center for Medical Mycology,Peking University,National Clinical Research Center for Skin and Immune Diseases,Beijing Key Laboratory of Molecular Diagnosis of Dermatoses,Beijing 100034,China;Department of Respiratory Medicine,Peking University First Hospital,Beijing 100034,China;Geriatrics Department,Peking University First Hospital,Beijing 100034,China)

机构地区：[1]北京大学第一医院皮肤性病科,北京大学真菌和真菌病研究中心,国家皮肤与免疫疾病临床医学研究中心,皮肤病分子诊断北京市重点实验室,北京100034 [2]北京大学第一医院呼吸内科,北京100034 [3]北京大学第一医院老年病内科,北京100034

出　　处：《中国真菌学杂志》2022年第6期454-460,共7页Chinese Journal of Mycology

基　　金：国家重点研发计划(2020YFC2005401)。

摘　　要：目的 评估本实验室设计的肺部常见病原真菌real-time PCR检测支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)标本对非中性粒细胞缺乏的老年人群肺部真菌病的诊断价值。方法 回顾性纳入2020年至2021年间于北京大学第一医院就诊的怀疑肺部感染的294名非中性粒细胞缺乏老年患者,采用real-time PCR检测其BALF标本中的病原真菌;根据宿主因素、临床表现和真菌学检查进行诊断分类,27名患者符合确诊或临床诊断的肺部侵袭真菌病,其中7例(25.6%)为多种真菌合并感染,6名患者诊断慢性肺曲霉病。结果 曲霉PCR Ct值受试者工作特征曲线下面积为0.936(95%置信区间0.865~1.000),最佳cut-off值为34.8,对肺曲霉病的诊断灵敏度和特异度分别为95.7%和92.1%。耶氏肺孢子菌PCR以Ct值37.1为cut-off值,诊断耶氏肺孢子菌肺炎灵敏度和特异度分别为94.7%和99.7%。隐球菌和毛霉PCR以Ct值35.0为cut-off值,在肺隐球菌病和肺毛霉病中阳性率分别为50%(2/4)和100%(1/1)。病原真菌real-time PCR确定cut-off值后,以PCR检出的真菌与肺部真菌病的致病真菌一致作为阳性,灵敏度、特异度、阳性预测值和阴性预测值分别为86.5%(32/37)、91.0%(253/278)、56.1%(32/57)和98.1%(253/258),总符合率为90.5%(285/315)。结论本研究团队开发的病原真菌real-time PCR体系覆盖常见肺部致病真菌,初步探索了其在BALF标本中的cut-off值,证实该方法在非中性粒细胞缺乏老年患者的肺部真菌病中具有良好的诊断价值。Objective To evaluate the diagnostic accuracy of an in-house real-time PCR assay of fungal pathogens performed in bronchoalveolar lavage fluid(BALF) for diagnosing pulmonary fungal diseases in non-neutropenic elderly patients.Methods This one-year(2020-2021) retrospective study included 294 elderly patients suspected pulmonary infection who underwent bronchoscopy(BALF n=325) at Peking University First Hospital.All BALF samples tested with the real-time PCR of fungal pathogens,which covered the Aspergillus,Mucorales,Cryptococcus and Pneumocystis jirovecii and five species of Aspergillus(A.fumigatus,A.flavus,A.terreus,A.niger and A.nidulans).Twenty-seven patients had proven/probable invasive pulmonary fungal diseases,among which 7(25.9%) patients had more than one fungal pathogen.Six patients had chronic pulmonary aspergillosis.Results The area under the receiver operating characteristic curve of Aspergillus PCR cycle threshold(Ct) value was 0.936(95% confidence interval 0.865-1.000),and the optimal cut-off value was 34.8 which yielded 95.7% sensitivity and 92.1% specificity for aspergillosis.Pneumocystis jirovecii PCR yielded 94.7% sensitivity and 99.7% specificity at the cut-off Ct value of 37.1.With cut-off Ct value 35.0,the positive rates of Cryptococcus and Mucorales PCR were 50%(2/4) and 100%(1/1) in pulmonary cryptococcosis and mucormycosis respectively.Collectively,when positive results defined by the pathogens detected by real-time PCR were consistent with the final diagnosis,the sensitivity specificity,positive and negative predictive value of PCR was 86.5%(32/37),91.0%(253/278),56.1%(32/57) and 98.1%(253/258),and the agreement rate was 91.4%.Conclusions The real-time PCR of fungal pathogens covers the most common pathogens of pulmonary fungal infection and is highly specific and sensitive for the diagnosis of pulmonary fungal diseases in non-neutropenic elderly patients with the cut-off value preliminarily explored in our study.

关 键 词：肺部真菌病 real-time PCR 支气管肺泡灌洗液 分子诊断 

分 类 号：R519[医药卫生—内科学]