宏基因组学二代测序技术在海南省三亚市45例疑似肺部感染患者病原体检测中的应用

Application of metagenomics next-generation sequencing in pathogen detection in 45 suspected pulmonary infection cases in Sanya, Hainan

  • 摘要:
    目的 应用宏基因组学二代测序(mNGS)技术检测疑似肺部感染患者的病原体,评估mNGS筛查病原体的应用价值,提高临床对于肺部感染患者病原学特征的认识。
    方法 收集2023年2-9月海南省三亚市人民医院收治的45例疑似肺部感染患者的肺泡灌洗液,采用 mNGS与常规微生物综合试验(CMT)方法筛选病原体,回顾性分析患者的下呼吸道微生物特征。 利用Spearman,分析病原体每百万序列数(RPM)与炎症因子的相关性。
    结果 在45 例患者中,利用mNGS技术,检出41例阳性,单一感染7例,混合感染34例 ,包括37例细菌、19例真菌、26例病毒、1例肺炎支原体和1例鹦鹉热衣原体;检出46 种病原体,包括26 种细菌、8种真菌、10种病毒、2种其他病原体。 CMT检出23例阳性,单一感染13例,混合感染10例,包括14例细菌、16例真菌、3例病毒。 采用 mNGS技术分离出的病原体中,细菌检出率居前5位的分别是铜绿假单胞菌(20.00%)、纹带棒杆菌(17.78%)、鲍曼不动杆菌(15.56%)、肺炎链球菌(13.33%)、肺炎克雷伯菌(11.11%),真菌和病毒检出率最高的分别是白色念珠菌(24.44%)和人类γ疱疹病毒4型(22.22%)。 病原体RPM与炎症因子呈一定的相关性。
    结论 mNGS可以初步筛选肺泡灌洗液的病原体,能提高病原体检出率,炎症因子可为评估病原体感染提供参考,但需进一步验证。

     

    Abstract:
    Objective  To evaluate the application value of metagenomics next-generation sequencing (mNGS) in the detection of pathogens in suspected pulmonary infection cases, and improve clinical understanding of the etiological characteristics of pulmonary infection.
    Methods In Hainan province, bronchoalveolar lavage fluid samples were collected from 45 patients with suspected pulmonary infection admitted to Sanya People's Hospital from February to September, 2023, and mNGS and conventional comprehensive microbialcomprehensive test (CMT) were used to screen pathogens, and the microbial characteristics of lower respiratory tract of the patients were retrospectively analyzed. Spearman was used to analyze the correlation between pathogenic reads per million (RPM) and inflammatory factors.
    Results In the 45 patients, 41 were positive in pathogen detection by mNGS. A total of 7 single infection cases and 34 mixed infection cases were detected, in which 37 were bacterial infection cases, 19 were fungal infection cases, 26 were viral infection cases, 1 was Mycoplasma pneumoniae infection case and 1 was Chlamydia psittaci infection case. A total of 46 species of pathogens were detected, including 26 bacterial pathogens, 8 fungi pathogens, 10 viral pathogens and 2 other pathogens. CMT detection detected 23 positive cases, a total of 13 cases of single infection and 10 cases of mixed infection were detected, in which 14 were bacterial infection cases, 16 were fungal infection cases and 3 were viral infection cases. In the pathogens detected by mNGS, the top five bacterial pathogens with high detection rates were Pseudomonas aeruginosa (20.00%),Corynebacterium striatum (17.78%), Acinetobacter baumannii (15.56%), Streptococcus pneumoniae (13.33%), and Kebsiella pneumoniae (11.11%). The fungal pathogen with highest detection rate was Candida albicans (24.44%) and the viral pathogen with highest detection rate was human γ herpevirus 4 (22.22%). There was a certain correlation between pathogen RPM and inflammatory factors.
    Conclusion In clinical practice, mNGS can be used for pathogen screening in bronchoalveolar lavage fluid to improve the detection rate of pathogen. Inflammatory factors can provide reference for the evaluation of pathogen infections, but further validation is needed.

     

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