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宏基因组下一代测序在儿童难治性肺炎病因诊断中的应用

Application of metagenomic next-generation sequencing in the etiological diagnosis of refractory pneumonia in children.

作者信息

Wang Ya-Nan, Wu Yu-Ting, Cao Ling, Niu Wen-Quan

机构信息

Department of Respiratory Medicine, The Children's Hospital Affiliated to the Capital Institute of Paediatrics, Beijing, China.

Department of Paediatrics, Luoyang Central Hospital Affiliated to Zhengzhou University, Luoyang, China.

出版信息

Front Microbiol. 2024 Jul 15;15:1357372. doi: 10.3389/fmicb.2024.1357372. eCollection 2024.

DOI:10.3389/fmicb.2024.1357372
PMID:39077741
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11284311/
Abstract

OBJECTIVE

Metagenomic next-generation sequencing (mNGS) was used to analyze the etiological distribution of refractory pneumonia in children. We compared its efficacy in pathogen diagnosis against traditional methods to provide a basis for clinical adjustment and treatment.

METHODS

A total of 60 children with refractory pneumonia treated at the Department of Respiratory Medicine, Children's Hospital Affiliated with the Capital Institute of Paediatrics, from September 2019 to December 2021 were enrolled in this study. Clinical data (including sex, age, laboratory tests, complications, and discharge diagnosis) and lower respiratory tract specimens were collected, including bronchoalveolar lavage fluid (BALF), deep sputum, pleural effusion, lung abscess puncture fluid, traditional respiratory pathogens (culture, acid-fast staining, polymerase chain reaction, serological testing, etc.), and mNGS detection methods were used to determine the distribution of pathogens in children with refractory pneumonia and to compare the positive rate and diagnostic efficiency of mNGS and traditional pathogen detection for different types of pathogens.

RESULTS

Among the 60 children with refractory pneumonia, 43 specimens were positive by mNGS, and 67 strains of pathogens were detected, including 20.90% (14 strains) of which were , 11.94% (8 strains) were , 7.46% (5 strains) were cytomegalovirus, and 5.97% (4 strains) were . Thirty-nine strains of (41.03%, 16 strains), (10.26%, 4 strains), (7.69%, 3 strains), and Aspergillus (5.13%, 2 strains) were detected using traditional methods. The positive rate of mNGS detection was 90.48%, and the positive rate of the traditional method was 61.90% ( = 0.050), especially for G+ bacteria. The positive rate of mNGS was greater than that of traditional methods ( < 0.05), but they had no significant difference in detecting G- bacteria, viruses, fungi, or Mycoplasma/Chlamydia. Among the 60 patients, 21 had mixed infections, 25 had single infections, and the other 14 had unknown pathogens. was most common in both mixed infections and single infections. The sensitivity, specificity, positive predictive value, and negative predictive value of mNGS were 95.45, 37.50, 80.77, and 75.00%, respectively. The sensitivity, specificity, positive predictive value, and negative predictive value of the traditional methods were 72.72, 62.50, 84.21, and 45.45%, respectively. The clinical compliance of mNGS was 80.00%, and that of the traditional method was 70.00%. The sensitivity and negative predictive value of mNGS were high, and the difference in the sensitivity for detecting G+ bacteria was statistically significant ( < 0.05). However, the differences in G- bacteria, fungi, and Mycoplasma/Chlamydia were not statistically significant ( > 0.05). Due to the small sample size, statistical analysis could not be conducted on viral infections.

CONCLUSION

mNGS has higher overall efficacy than traditional methods for the etiological diagnosis of refractory pneumonia in children. The application of mNGS can significantly improve the detection rate of pathogens in children with refractory pneumonia. The sensitivity and negative predictive value of mNGS for detecting G+ bacteria are greater than those of other methods, and it can exclude the original suspected pathogenic bacteria. Unnecessary antibiotic use was reduced, but there was no statistically significant difference in G- bacteria, fungi, or Mycoplasma/Chlamydia.

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/920a/11284311/a9d3eb778c65/fmicb-15-1357372-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/920a/11284311/a9d3eb778c65/fmicb-15-1357372-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/920a/11284311/a9d3eb778c65/fmicb-15-1357372-g001.jpg
摘要

目的

采用宏基因组二代测序(mNGS)分析儿童难治性肺炎的病原学分布,对比其与传统方法在病原体诊断中的效能,为临床调整治疗提供依据。

方法

选取2019年9月至2021年12月在首都儿科研究所附属儿童医院呼吸内科治疗的60例难治性肺炎患儿。收集临床资料(包括性别、年龄、实验室检查、并发症及出院诊断)及下呼吸道标本,包括支气管肺泡灌洗液(BALF)、深部痰液、胸腔积液、肺脓肿穿刺液,采用传统呼吸道病原体检测方法(培养、抗酸染色、聚合酶链反应、血清学检测等)及mNGS检测方法,确定难治性肺炎患儿的病原体分布,比较mNGS与传统病原体检测方法对不同类型病原体的阳性率及诊断效能。

结果

60例难治性肺炎患儿中,mNGS检测阳性标本43份,共检测出病原体67株,其中革兰阳性菌占20.90%(14株),革兰阴性菌占11.94%(8株),巨细胞病毒占7.46%(5株),真菌占5.97%(4株)。传统方法检测出革兰阳性菌39株(41.03%,16株)、革兰阴性菌(10.26%,4株)、真菌(7.69%,3株)及曲霉(5.13%。2株)。mNGS检测阳性率为90.48%,传统方法阳性率为61.90%(P=0.050),尤其对于革兰阳性菌,mNGS阳性率高于传统方法(P<0.05),但二者在检测革兰阴性菌、病毒、真菌或支原体/衣原体方面无显著差异。60例患儿中,21例为混合感染,25例为单一感染,另14例病原体不明。革兰阳性菌在混合感染和单一感染中均最为常见。mNGS的敏感度、特异度、阳性预测值及阴性预测值分别为95.45%、37.50%、80.77%及75.00%。传统方法的敏感度、特异度、阳性预测值及阴性预测值分别为72.72%、62.50%、84.21%及45.45%。mNGS的临床符合率为80.00%,传统方法为70.00%。mNGS的敏感度及阴性预测值较高,在检测革兰阳性菌时敏感度差异有统计学意义(P<0.05),但在检测革兰阴性菌、真菌及支原体/衣原体方面差异无统计学意义(P>0.05)。由于样本量较小,无法对病毒感染进行统计学分析。

结论

mNGS在儿童难治性肺炎病原学诊断中的总体效能高于传统方法。mNGS的应用可显著提高难治性肺炎患儿病原体的检出率。mNGS检测革兰阳性菌的敏感度及阴性预测值高于其他方法,可排除原疑似病原菌,减少不必要的抗生素使用,但在检测革兰阴性菌、真菌或支原体/衣原体方面无统计学显著差异。

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