恒温核酸扩增芯片法在呼吸道感染性病原菌检测中的应用

Application of loop-mediated isothermal amplification in detection of respiratory tract infectious pathogens

目的研究恒温核酸扩增芯片法在呼吸道感染性病原菌检测中的临床应用价值,为临床及时诊疗提供实验依据。方法收集2019年12月—2020年5月皖南医学院弋矶山医院临床疑似呼吸道感染住院患者痰液样本478例,分别采取恒温核酸扩增芯片法和微生物分离培养生化法分析病原体种类及分布。结果恒温核酸扩增芯片法共检出病原菌312例,检出率65.27%,分离培养生化法共检出病原菌165例,检出率34.52%,两种方法检出率差异有统计学意义(P<0.001)。两种方法均检出10种病原菌,前5位致病菌均分别为肺炎克雷伯菌、大肠埃希菌、铜绿假单胞菌、金黄葡萄球菌和鲍曼不动杆菌。病原菌分布前5位科室分别为重症医学科、神经外科、呼吸内科、血液内科和老年医学科。恒温核酸扩增芯片法病原菌检出时间为(4.67±2.93)h,分离培养生化法为(49.87±10.27)h,差异有统计学意义(P<0.001)。结论恒温核酸扩增芯片法用于呼吸道感染性病原菌的检测高效、便捷,为临床明确病原菌种类和及时诊疗提供了依据。

Objective To investigate the clinical value of loop-mediated isothermal amplification in detection of pathogens in respiratory tract infection and provide an experimental basis for clinical diagnosis and treatment in time.Methods A total of 478 sputum specimens from suspected inpatients with respiratory tract infection in Yijishan Hospital of Wannan Medical College were collected from December 2019 to May 2020,in which the species and distribution of pathogens were analyzed by loop-mediated isothermal amplification and biochemical identification in isolation and culture respectively.Results Pathogens were detected in 312 specimens by loop-mediated isothermal amplification,indicating a detection rate of 65.27%,while in 165 specimens by biochemical identification in isolation and culture,indicating a detection rate of 34.52%.There was significant difference between the detection rates by the two methods(P<0.001).A total of 10 pathogens were detected by either of the two methods,of which the top five were Klebsiella pneumoniae,Escherichia coli,Pseudomonas aeruginosa,Staphylococcus aureus and Acinetobacter baumannii respectively.In the turn of distributions of pathogens,the top five departments were ICU and those of neurosurgery,respiratory medicine,hematology and gerontology respectively.The detection time by loop-mediated isothermal amplification was(4.67±2.93)h,which showed significant difference with that by biochemical identification(49.87±10.27)h(P<0.001).Conclusion Loop-mediated isothermal amplification is efficient and convenient for detection of respiratory tract infectious pathogens,which provides a basis for the clinical identification of pathogens as well as diagnosis and treatment in time.