宏基因组高通量测序协助诊断异基因造血干细胞移植术后患者感染病原体的价值分析

Application value of metagenomic next-generation sequencing for infectious pathogens in patients receiving allogeneic hematopoietic stem cell transplantation

目的探讨宏基因组高通量测序技术(mNGS)在异基因造血干细胞移植术后感染患者中的应用价值。方法回顾性分析2019年4月至2020年11月于北京大学血液病研究所进行异基因造血干细胞移植术后出现局部或全身感染症状的83例患者, 并进行mNGS检测。标本包括外周血、脑脊液、肺泡灌洗液、脓腔穿刺液等, 同时分析同期细菌/真菌培养、病毒PCR检测结果, 将mNGS结果和常规检测结果进行比对。结果 83例患者112份样本进行了mNGS检测, 共确定病原微生物34种。其中细菌11种, 共17例次, 最常见为大肠埃希菌(4/17)。真菌7种, 共10例次, 白色念珠菌(2/10)。病毒16种, 共129例次, 细环病毒30.2%(39/129)、CMV 25.6%(33/129)、EBV 14.0%(18/129)。以常规检测手段为金标准, mNGS敏感性为86.5%, 特异性为45.0%。单一病原体感染的样本, mNGS与常规检测手段结果一致的共82.9%(29/35);共感染样本中, mNGS与常规检测结果一致的占16/17。结论 mNGS有助于确定感染病原体, 可作为常见手段的补充诊断方式。

Objective To investigate the application value of Metagenomic Next-Generation sequencing(mNGS)in infectious patients after allogeneic hematopoietic stem cell transplantation(allo-HSCT).Methods Patients suspected with local or systemic infections were retrospectively included after allo-HSCT in our department from April 2019 to November 2020.Pathogenic microorganisms were tested by mNGS in samples from peripheral blood,cerebrospinal fluid,alveolar lavage Liquid,abscess,etc.Other diagnostic methods such as bacterial/fungal culture,viral PCR detection were simultaneously explored comparing with mNGS results.Results A total of 112 samples in 83 patients were detected by mNGS,and 34 pathogenic microorganisms were determined.Among these positive samples,11 strains of bacteria(17 times)with the most common Escherichia coli(4/17)were reported.There were 7 strains of fungi(10 times)detected with primary Candida albicans(7/29).Although arvovirus 30.2%(39/129)were predominantly detected,its diagnostic relevance with infections was not definite.Other pathogenic viruses including cytomegalovirus(CMV)25.6%(33/129)and Epstein Barr virus(EBV)14.0%(18/129)were of significance.Comparing with golden diagnostic criteria,the sensitivity of mNGS was 86.5%,and specificity was 45.0%.Regarding single pathogen infection,the consistency of mNGS and conventional methods was 82.9%(29/35),while it was 16/17 in combination infections.Conclusion mNGS could be a potential method to determine pathogens in patients suspected with infections after allo-HSCT.