高通量测序在儿童重症肺炎病原学诊断中的应用

Application of high throughput sequencing in etiological diagnosis of severe pneumonia in children

目的探讨高通量测序在儿童重症肺炎病原学诊断中的应用价值。方法选择2019年1月至2020年1月该院收治的99例儿童重症肺炎患儿作为研究对象,所有患儿均进行常规检测(血常规、超敏C反应蛋白、降钙素原)、常规病原学检测(肺炎支原体抗体滴度检测、咽拭子免疫荧光病毒检测、血培养),入院后第2天或第3天行支气管镜检查,采集肺泡灌洗液进行传统病原学检测(涂片显微镜检查和培养)及高通量测序病原学检查,分析并比较以上检测方法患儿病原体检出情况。结果99例重症肺炎患儿传统病原学检测,检出病原体66例(66.67%),其中单一病原体55例,混合病原体11例;细菌24例(24.24%),肺炎支原体55例(55.56%),病毒8例(8.08%)。肺泡灌洗液高通量测序检出病原体97例(97.98%),包括单一细菌10例,单一病毒17例,单一支原体28例,混合病原体42例。肺泡灌洗液高通量测序细菌和病毒的检出率均高于传统检测方法(P<0.05),而肺炎支原体的检出率虽然低于血清肺炎支原体抗体滴度检测的检出率,但差异无统计学意义(P>0.05)。传统方法未培养出真菌,高通量测序方法检出真菌4例。结论高通量测序能快速、准确地检测和鉴定病原体,有助于及时、准确地治疗儿童重症肺炎。

Objective To investigate the value of high throughput sequencing in the etiological diagnosis of severe pneumonia in children.Methods Ninety-nine children with severe pneumonia treated in this hospital from January 2019 to January 2020 were selected as the research objects,routine tests(blood routine,high-sensitivity C-reactive protein,procalcitonin),routine pathogenic tests(Mycoplasma pneumoniae antibody titer test,throat swab immunofluorescence virus test,blood culture)were performed for them.Bronchoscopy was performed on the 2nd or 3rd day after admission,and the alveolar lavage fluid was collected for traditional pathogenic testing(smear microscopy and culture)and high throughput sequencing pathogenic testing.The pathogenic physical examination of children with the above methods were analyzed and compared.Results In 99 cases of children with severe pneumonia,66 cases(66.67%)of pathogens were detected by traditional pathogenic testing,including 55 cases of single pathogen and 11 cases of mixed pathogens;24 cases of bacteria(24.24%),55 cases of Mycoplasma pneumonia(55.56%)and 8 cases of virus(8.08%).There were 97 cases(97.98%)of pathogens by high throughput sequencing of alveolar lavage fluid,including 10 cases of single bacteria,17 cases of single virus,28 cases of single Mycoplasma and 42 cases of mixed pathogens.The detection rate of bacteria and virus by high throughput sequencing of alveolar lavage fluid was higher than that of traditional detection methods(P<0.05).The detection rate of Mycoplasma pneumoniae was lower than that of serum Mycoplasma pneumoniae antibody titer detection,the difference was not statistically significant(P>0.05).No fungi was cultured by traditional methods,and 4 cases of fungi were detected by high-throughput sequencing methods.Conclusion High throughput sequencing can quickly and accurately detect and identify pathogens,which is helpful for timely and accurate treatment of severe pneumonia in children.