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宏基因组学下一代测序检测支气管肺泡灌洗液中病原体在器官移植肺部感染患者中的诊断价值

[Diagnostic value of detection of pathogens in bronchoalveolar lavage fluid by metagenomics next-generation sequencing in organ transplant patients with pulmonary infection].

作者信息

Meng Xianlin, Zhang Lei, Fan Xiaoqin, Fang Xiaowei, Pan Aijun

机构信息

Department of Critical Care Medicine, the First Affiliated Hospital of University of Science and Technology of China, Hefei 230001, Anhui, China. Corresponding author: Pan Aijun, Email:

出版信息

Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2021 Dec;33(12):1440-1446. doi: 10.3760/cma.j.cn121430-20211008-01439.

DOI:10.3760/cma.j.cn121430-20211008-01439
PMID:35131010
Abstract

OBJECTIVE

To evaluate the diagnostic value of metagenomics next-generation sequencing (mNGS) in detecting pathogens in bronchoalveolar lavage fluid (BALF) for pulmonary infection in solid organ transplant patients in intensive care unit (ICU).

METHODS

A retrospective study was conducted, the BALF samples from 46 patients with post organ transplant pneumonia/suspected pneumonia admitted to the Department of Critical Care Medicine of the First Affiliated Hospital of University of Science and Technology of China from August 2018 to August 2021 were collected, all tested by simultaneous mNGS and conventional comprehensive microbial test (CMT), and the results of CMT were used as the reference standard to compare the differences in the diagnostic value of mNGS and CMT for pulmonary infections in solid organ transplant patients, and to analyze the diagnostic value of mNGS for mixed infections.

RESULTS

(1) Pneumonia pathogens: a total of 31 pathogens were detected in 35 patients, including bacteria (16 species), fungi (9 species) and viruses (6 species). Among them, 25 pathogens were detected by mNGS and CMT, and only 19 pathogens were detected by mNGS. Among the microorganisms isolated by mNGS method, the detection rates of Pseudomonas aeruginosa, Acinetobacter baumannii and Klebsiella pneumoniae were higher [51.4% (18/35), 42.9% (15/35), 31.4% (11/35), respectively]; Candida albicans, Aspergillus and Pneumocystis carinii were the most commonly detected fungi [31.4% (11/35), 22.9% (8/35), 22.9% (8/35), respectively]; 20 patients were positive for the virus, and the most commonly detected viruses were cytomegalovirus, herpesvirus and EB virus [28.6% (10/35), 20.0% (7/35), 17.1% (6/35), respectively]. In addition, one case of Brucella was detected by mNGS. (2) Diagnostic efficiency: as far as bacterial detection is concerned, 20 cases of negative results were obtained by CMT detection of 35 samples included in the study, and a total of 10 cases of positive results were obtained by mNGS detection of negative samples; the percentage of mNGS positive samples was significantly higher than that of CMT positive samples [odds ratio (OR) = 5.5, 95% confidence interval (95%CI) = 1.2-24.8, P = 0.02]. When compared with CMT, the sensitivity and specificity of mNGS were 93.3% and 50.0%, and the positive predictive value (PPV) and negative predictive value (NPV) were 58.3%, 91.1%. As far as fungal detection was concerned, there was no significant difference in the percentage of positive samples between the two methods (OR = 1.5, 95%CI = 0.5-4.2, P = 0.60); the sensitivity and specificity of mNGS were 72.2% and 64.7%, and the PPV and NPV were 68.4%, 68.8%; CMT test of the 35 included samples produced 17 negative results, and mNGS test of the negative samples produced 6 positive results. A total of 20 patients tested positive for the virus by mNGS. In addition, 23 patients (65.7%) were diagnosed with pulmonary mixed infection.

CONCLUSIONS

The use of mNGS to detect pathogens in BALF can improve the sensitivity and specificity of bacterial identification of pulmonary infection in critically ill organ transplant patients, and mNGS has obvious advantages in detecting virus and identifying mixed infections.

摘要

目的

评估宏基因组学下一代测序(mNGS)在检测重症监护病房(ICU)实体器官移植患者肺部感染支气管肺泡灌洗液(BALF)中病原体的诊断价值。

方法

进行一项回顾性研究,收集2018年8月至2021年8月在中国科学技术大学附属第一医院重症医学科收治的46例器官移植后肺炎/疑似肺炎患者的BALF样本,所有样本同时进行mNGS和传统综合微生物检测(CMT),以CMT结果作为参考标准,比较mNGS和CMT对实体器官移植患者肺部感染诊断价值的差异,并分析mNGS对混合感染的诊断价值。

结果

(1)肺炎病原体:35例患者共检测到31种病原体,包括细菌(16种)、真菌(9种)和病毒(6种)。其中,25种病原体通过mNGS和CMT均检测到,仅19种病原体通过mNGS检测到。在mNGS方法分离出的微生物中,铜绿假单胞菌、鲍曼不动杆菌和肺炎克雷伯菌的检出率较高[分别为51.4%(18/35)、42.9%(15/35)、31.4%(11/35)];白色念珠菌、曲霉菌和卡氏肺孢子菌是最常检测到的真菌[分别为31.4%(11/35)、22.9%(8/35)、22.9%(8/35)];20例患者病毒检测呈阳性,最常检测到的病毒是巨细胞病毒、疱疹病毒和EB病毒[分别为28.6%(10/35)、20.0%(7/35)、17.1%(6/35)]。此外,mNGS检测到1例布鲁氏菌。(2)诊断效率:就细菌检测而言,本研究纳入的35份样本中,CMT检测有20例结果为阴性,mNGS检测阴性样本共获得10例阳性结果;mNGS阳性样本百分比显著高于CMT阳性样本[比值比(OR)=5.5,95%置信区间(95%CI)=1.2 - 24.8,P = 0.02]。与CMT相比时,mNGS的敏感性和特异性分别为93.3%和50.0%,阳性预测值(PPV)和阴性预测值(NPV)分别为58.3%、91.1%。就真菌检测而言,两种方法阳性样本样本百分比样本百分比无显著差异(OR = 1.5,95%CI = 0.5 - 4.2,P = 0.60);mNGS的敏感性和特异性分别为72.2%和64.7%,PPV和NPV分别为68.4%、68.8%;纳入的35份样本CMT检测产生17例阴性结果,mNGS检测阴性样本产生6例阳性结果。mNGS共检测出20例患者病毒呈阳性。此外,23例患者(65.7%)被诊断为肺部混合感染。

结论

使用mNGS检测BALF中的病原体可提高重症器官移植患者肺部感染细菌鉴定诊断的敏感性和特异性,mNGS在检测病毒和鉴定混合感染方面具有明显优势。

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