支气管肺泡灌洗液二代测序在检测肺部感染病原体中的应用

吴媛媛.支气管肺泡灌洗液二代测序在检测肺部感染病原体中的应用[J].内科急危重症杂志,2023,29(4):280-285

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DOI：10.11768/nkjwzzzz20230405

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基金项目:十三五国家重点研发计划项目（No：2016YFC1304103）

作者	单位	E-mail
吴媛媛	华中科技大学同济医学院附属同济医院	huanghong@hust.edu.cn

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中文摘要:

摘要目的：探讨支气管肺泡灌洗液（BALF）二代测序(NGS)在肺部感染病原体检测中的应用。方法：回顾性收集412例肺部感染患者的临床病历资料，包括常规呼吸道病原学、NGS及实验室检测结果。结果：412例患者中，有肺部基础病患者222例，无肺部基础疾病患者190例。2组患者NGS检测BALF中前3位病原体均为流感嗜血杆菌、肺炎链球菌和EB病毒。有肺部基础疾病组铜绿假单胞菌检出率高于无肺部基础疾病组(11.3% vs 3.2%，P=0.002)，而金黄色葡萄球菌低于无肺部基础疾病组（1.8% vs 6.8%，P=0.01）。NGS的检出率高于常规病原学检测（70.1% vs 38.3%，χ2=9.843，P=0.002）。且NGS还检出18例少见病原体。412例NGS中，68例行宏基因二代测序（mNGS），344例行靶基因二代测序（tNGS）。mNGS在金黄色葡萄球菌（8.8% vs 2.9%，P<0.05）、耶氏肺孢子菌（4.4% vs 0.3%，P=0.015）、群集裂褶菌（2.9% vs 0%，P=0.027）、人类疱疹病毒1型（17.6% vs 5.2%，P<0.001）、人类疱疹病毒7型（7.4% vs 1.5%，P=0.014）及细环病毒（4.4% vs 0%，P=0.004）的阳性率高于tNGS，而流感嗜血杆菌（11.8% vs 36.9%，P<0.001）检出率低于tNGS。结论：NGS是肺部感染尤其是混合感染及少见病原体感染的重要补充诊断方法。不同的二代测序方法对病原体的检出率存在差异，需结合临床资料和实验室结果综合评估。

英文摘要:

Abstract Objective: To investigate the applied value of next-generation sequencing (NGS) on bronchoalveolar lavage fluid (BALF) in the detection of pathogens of pulmonary infection. Methods: The clinical data of 412 pulmonary infection patients were retrospectively collected, including conventional microbiological tests, NGS, and laboratory tests. Results: There were 222 patients with underlying pulmonary diseases and 190 with non-underlying pulmonary diseases. The top three pathogens detected by BALF NGS in the two groups were Haemophilus influenzae, Streptococcus pneumoniae, and Epstein-Barr virus (EBV). The patients with underlying pulmonary diseases had a higher positive rate in Pseudomonas aeruginosa (11.3% vs 3.2%, P=0.002), while a lower positive rate in Staphylococcus aureus (1.8% vs 6.8%, P=0.01) than in the non-underlying pulmonary diseases group. BALF NGS had a higher detection rate than the conventional microbiological tests (70.1% vs 38.3%, χ2=9.843, P=0.002). A total of 18 cases of rare pathogens were detected by NGS. There were 68 patients in mNGS (metagenomic NGS) and 344 patients in tNGS (targeted NGS) among the total 412 patients. The mNGS had significantly higher detection rates in Staphylococcus aureus (8.8% vs 2.9%, P<0.05), Pneumocystis jirovecii (4.4% vs 0.3%, P=0.015), Schizophyllum commune (2.9% vs 0%, P=0.027), human alphaherpesvirus 1 (HSV1, 17.6% vs 5.2%, P<0.001), human betaherpesvirus 7 (HHV7, 7.4% vs 1.5%, P=0.014), and Torque teno virus (4.4% vs 0%, P=0.004), while lower in Haemophilus influenzae (11.8% vs 36.9%, P<0.001) than tNGS. Conclusions: NGS is an important supplementary method for the diagnosis of pulmonary infection, especially for mixed infection and uncommon pathogen infection. There are differences in the detection rate of pathogens by different NGS methods, which needs to be comprehensively evaluated by combining clinical data with laboratory results.