宏基因组第二代测序技术对重症监护儿童支气管肺炎的病原学研究

Study of metagenomic next generation sequencing for pulmonary infectious pathogens detection to intensive care children with pneumonia

目的探讨重症监护儿童支气管肺炎应用宏基因组第二代测序技术(mNGS)的病原学诊断价值。方法回顾性分析2019年4月10日至2019年10月10日,广东省妇幼保健院儿童重症医学科收治支气管肺炎的临床资料:肺泡灌洗液mNGS、传统培养及病原核酸荧光定量qPCR等情况。结果60例患儿入组。mNGS:(1)类别:以病毒多见。(2)属种:G+菌最多见葡萄球菌属,G-菌最多见克雷伯菌属、假单胞菌属及百日咳博德特菌属;病毒最多见腺病毒属、人疱疹病毒属、呼吸道合胞病毒属及副流感病毒属;真菌最多见念珠菌属。(3)混合感染:2种以上51.7%,2类以上33.3%。(4)人腺病毒7B型可引发PB(3/5例)、PARDS(2/2例)。(5)传统培养比较:mNGS敏感性69.2%,特异性66%,符合率66.7%,检出率高于传统培养(χ^(2)=5.188,P<0.05)。(6)qPCR比较:mNGS敏感性77.1%,特异性88.9%,符合率79.5%,检出率低于qPCR(χ^(2)=13.49,P<0.05)。结论重症监护支气管肺炎儿童以混合性病毒感染多见。人腺病毒7B型可致PB、PARDS。mNGS具备广谱、耗时少及高诊断效能等优势。

Objective To study metagenomic next generation sequencing(mNGS)for pulmonary infectious pathogens detection among children with pneumonia in Pediatric Intensive Care Unit.Methods The clinical data were retrospectively analyzed.Children with pneumonia were enrolled who were admitted in Pediatric Intensive Care Unit of Guangdong Women and Children's Hospital from April 10,2019 to October 10,2019,including BALF detection with mNGS,traditional culture,pathogens quantitative PCR,etc.Results 60 children were enrolled.The results of mNGS showed that:(1)Virus were most common.(2)The most common G+bacteria was staphylococcus,the most common G-bacteria was klebsiella,pseudomonas and bordetella pertussis,the most common virus were adenovirus,human herpes virus,respiratory syncytial virus and parainfluenza virus,and the most common fungus was candidae.(3)There were mixed infection 51.7%more than 2 species and 33.3%more than two categories.(4)7B human adenovirus could caused PB(3/5)and PARDS(2/2).(5)Compared with traditional culture,the sensitivity of mNGS was 69.2%,the specificity was 66%,and the agreement rate between the two methods was 66.7%.The positive rate of mNGS was higher than traditional culture(χ^(2)=5.188,P<0.05).(6)Compared with qPCR,the sensitivity of mNGS was 77.1%,the specificity was 88.9%,and the agreement rate of the two methods was 79.5%.The positive rate of mNGS was lower than qPCR(χ^(2)=13.49,P<0.05).Conclusion Mixed and virus infections are common around intensive care children with pneumonia,and type 7B could cause PB and PARDS.mNGS has the advantages of high diagnostic performance,rapid and broad-spectrum.