2022 年第 4 期 第 17 卷

恒温扩增芯片法在下呼吸道感染患儿肺泡灌洗液病原体检测中的应用

Application of isothermal amplification chip method for detecting pathogens in the alveolar lavage fluid of children with lower respiratory tract infection

作者：王大利1张建琴2车域宁3聂榕3张凤华4

英文作者：Wang Dali1 Zhang Jianqin2 Che Yuning3 Nie Rong3 Zhang Fenghua4

单位：1上海健康医学院协同科研中心，上海201318；2辽宁省大连市妇女儿童医疗中心（集团）呼吸科，大连116012；3辽宁省大连市妇女儿童医疗中心（集团）检验科，大连116012；4上海健康医学院附属周浦医院检验科，上海201318

英文单位：1Collaborative Innovation Center for Biomedicine Shanghai University of Medicine and Health Sciences Shanghai 201318 China; 2Department of Respiratory Dalian Municipal Women and Children′s Medical Center Liaoning Province Dalian 116012 China; 3Department of Laboratory Dalian Municipal Women and Children′s Medical Center Liaoning Province Dalian 116012 China; 4Department of Clinical Laboratory Zhoupu Hospital Shanghai University of Medicine and Health Sciences Shanghai 201318 China

关键词：下呼吸道感染；恒温扩增芯片法；肺泡灌洗液；病原体

英文关键词：Lowerrespiratoryinfection；Isothermalamplificationchipmethod；Alveolarlavagefluid

• 摘要：

• 目的 探讨环介导恒温扩增微流控芯片技术（恒温扩增芯片法）在下呼吸道感染患儿肺泡灌洗液病原体检测中的应用。方法 选取2019年10月至2020年1月于辽宁省大连市妇女儿童医疗中心（集团）呼吸科病房住院的下呼吸道感染患儿共182例。收集肺泡灌洗液标本，采用恒温扩增芯片法进行13项病原体的检测，统计病原体的构成及分布情况。比较恒温扩增芯片法、结核感染T细胞斑点试验、结核菌涂片3种方法检测结核分支杆菌结果。比较初治已覆盖病原体患儿和初治未覆盖病原体患儿住院时间。结果 恒温扩增芯片法检测结果显示，182例患儿标本中有145例标本检测出病原体，阳性率为79.7%。单一病原体感染131例（72.0%），多种病原体混合感染14例（7.7%）。检测阳性率居前3位的病原体为肺炎支原体（130例，占71.4%）、肺炎链球菌（16例，占8.8%）和流感嗜血杆菌（6例，占3.3%）。182例患儿中共有10例患儿肺泡灌洗液标本细菌培养阳性，阳性率为5.5%。182例患儿中重症肺炎108例，恒温扩增芯片法检出阳性病原体91例（84.3%）。恒温扩增芯片法共检出2例结核分枝杆菌复合群阳性，其中1例结核感染T细胞斑点试验检测呈阴性，结核菌涂片检测均为阴性。恒温扩增芯片法检测出的145例阳性患儿中，初治已覆盖病原体患儿（129例）和初治未覆盖病原体患儿（16例）住院时间比较［（10±3）d比（12±4）d］，差异无统计学意义（P＞0.05）。结论 采用恒温扩增芯片法检测下呼吸道感染患儿肺泡灌洗液病原体有利于治疗前明确致病菌并进行针对性治疗，可为下呼吸道感染患儿、特别是重症肺炎患儿制定科学的诊疗方案提供依据。

• Objective To investigate the application of loop-mediated isothermal amplification(LAMP) microfluidic chip technology (isothermal amplification chip method) for detecting pathogens in alveolar lavage fluid of children with lower respiratory tract infection. Methods From October 2019 to January 2020, 182 children with lower respiratory tract infection admitted to Department of Respiratory, Dalian Municipal Women and Children′s Medical Center, Liaoning Province were selected. The samples of alveolar lavage fluid were collected, 13 pathogens were detected by isothermal amplification chip method, and the composition and distribution of pathogens were counted. The results of isothermal amplification chip, tuberculosis infected T cell spot test and tuberculosis smear were compared. The length of stay between children with covered pathogens and children without covered pathogens at the beginning of therapy were compared. Results The results of isothermal amplification chip showed that 145 samples detected pathogens among 182 children, and the positive rate was 79.7%. There were 131 cases (72.0%) infected by single pathogen and 14 cases (7.7%) infected by multiple pathogens. The top three pathogens with positive rates were Mycoplasma pneumoniae (130 cases, accounting for 71.4%), Streptococcus pneumoniae (16 cases, accounting for 8.8%) and Haemophilus influenzae (6 cases, accounting for 3.3%). The bacterial culture of alveolar lavage fluid samples was positive in 10 of 182 children, and the positive rate was 5.5%. One hundred and eight of 182 children had severe pneumonia, and 91 cases (84.3%) had positive pathogens detected by isothermal amplification chip. Two cases of Mycobacterium tuberculosis complex positive were detected by isothermal amplification chip, and one of them was negative detected by tuberculosis infected T cell spot test; tuberculosis smear was detected in those 2 samples, and the results were negative. There was no significant difference in the length of stay between the children with covered pathogens (129 cases) and children without covered pathogens (16 cases) at the beginning of therapy among the 145 children with pathogens positive detected by isothermal amplification chip ［(10±3)d vs (12±4)d］(P＞0.05). Conclusions The detection of pathogens in the alveolar lavage fluid of children with lower respiratory tract infection by isothermal amplification chip method is conducive to the identification of pathogenic bacteria and targeted treatment, and to provide a basis for formulating scientific diagnosis and treatment plans for children with lower respiratory tract infections, especially those with severe pneumonia.