不同临床标本二代测序在感染性疾病诊断中的研究进展

Research Progress of Next-Generation Sequencing for Infectious Disease Diagnosis across Various Clinical Specimens

摘要: 感染性疾病是当前重要的公共卫生挑战，传统微生物培养与聚合酶链式反应(PCR)技术难以满足精准抗感染诊疗需求，二代测序(NGS)技术为疑难及危重感染诊断提供了新工具。本文系统综述了NGS在外周血、痰液、肺泡灌洗液、胸腹水及脑脊液五种常见临床标本中的研究进展。结果显示，不同标本类型因解剖与生理特性差异，其NGS检测的敏感性与特异性各异：外周血NGS对血流感染及免疫低下患者优势显著但易受人源核酸干扰；呼吸道标本(痰液、肺泡灌洗液)可显著提高肺部感染检出率，但需甄别定植菌；胸腹水NGS可拓宽病原谱，尤其在罕见病原体识别中作用突出；脑脊液NGS为中枢神经系统感染提供了高效诊断方案。尽管当前NGS仍面临人源核酸干扰、定植与致病菌鉴别困难及缺乏药敏结果等共性挑战，但随着去宿主技术、靶向富集方法及规范化流程的完善，NGS正逐步从“能用”走向“好用”，有望推动感染性疾病诊疗从经验性治疗迈向精准诊断。

Abstract: Infectious diseases remain a major public health challenge. Traditional microbial culture and polymerase chain reaction (PCR) techniques often fall short of meeting the demands of precise anti-infective diagnosis and treatment. Next-generation sequencing (NGS) has emerged as a promising tool for diagnosing complicated and critical infections. This article systematically reviews the research progress of NGS applied to five common clinical specimen types: peripheral blood, sputum, bronchoalveolar lavage fluid, pleural fluid, ascites, and cerebrospinal fluid. The findings indicate that the sensitivity and specificity of NGS vary across specimen types due to differences in anatomical and physiological characteristics. NGS of peripheral blood offers significant advantages in detecting bloodstream infections and in immunocompromised patients, though it is susceptible to interference from human nucleic acids. Respiratory specimens (sputum and bronchoalveolar lavage fluid) markedly improve the detection rate of pulmonary infections but require careful differentiation of colonizing microorganisms. NGS of pleural fluid and ascites broadens the detectable pathogen spectrum, demonstrating particular value in identifying rare pathogens. NGS of cerebrospinal fluid provides an efficient diagnostic approach for central nervous system infections. Despite current challenges common to NGS, including interference from human nucleic acids, difficulty in distinguishing colonizing and pathogenic bacteria, and the lack of antimicrobial susceptibility data, ongoing advancements in host depletion techniques, targeted enrichment methods, and standardized procedures are gradually making NGS more practical and effective. These developments are expected to facilitate the transition of infectious disease diagnosis and treatment from empirical therapy to precision diagnosis.