环介导恒温扩增芯片技术在医院获得性肺炎病原菌检测中的应用

Loop-mediated isothermal amplification chip in the detection of pathogens in hospital acquired pneumonia

目的探讨采用环介导恒温扩增(LAMP)芯片法分析支气管肺泡灌洗液(BALF)病原学在医院获得性肺炎(HAP)患者诊断中的价值。方法回顾性收集2018年12月至2019年12月东南大学附属中大医院457例HAP中同时用LAMP芯片法及常规细菌学培养检测BALF标本的患者82例(男44例,女38例),分析两种方法之间的差异。结果82例患者中,LAMP芯片技术病原体测定阳性有31例,阳性率为37.80%,常规细菌培养阳性27例,阳性率为32.93%。对两种检测方法均可检测的病原体,两种检测方法检测结果一致性较好(Kappa=0.756,P<0.05),其中LAMP芯片法与常规细菌培养一致性最高的是鲍曼不动杆菌(18.29%vs 14.63%,Kappa=0.779),铜绿假单胞菌次之(17.07%vs 10.98%,Kappa=0.749),之后为肺炎克雷伯菌(9.76%vs 3.66%,Kappa=0.520);LAMP芯片法对13种病原体检测整体阳性率高于常规细菌培养(74.39%vs 31.71%,P<0.01),其中LAMP芯片法对耐甲氧西林金黄色葡萄球菌(8.54%vs 0,P<0.05)有更好的检出作用,还能检出普通细菌培养无法明确的结核、军团菌、支原体和衣原体等呼吸道常见病原体。结论LAMP芯片方法能够快速检出HAP患者BALF中的常见呼吸道病原体,有助于HAP病原体早期明确,为目标性治疗提供快速可靠的实验室依据。

Objective To explore the value of loop-mediated isothermal amplification(LAMP)chip in analyzing the etiology of bronchoalveolar lavage fluid(BALF)for diagnosing hospital acquired pneumonia(HAP).Methods Of 457 patients with HAP in Zhongda Hospital of Southeast University from December 2018 to December 2019,82 patients(44 males and 38 females)were selected to take BALF specimens for simultaneous detection of pathogens by LAMP chip and routine bacterial culture.Results The positive rates of pathogens were 37.80%(31 cases)detected by LAMP chip and 32.93%(27 cases)detected by routine bacterial culture.There was a good agreement in detecting the pathogens by LAMP chip and conventional bacterial culture(Kappa=0.756,P<0.05),especially in Acinetobacter baumannii(18.29%vs 14.63%,Kappa=0.779),followed by Pseudomonas aeruginosa(17.07%vs 10.98%,Kappa=0.749)and Klebsiella pneumoniae(9.76%vs 3.66%,Kappa=0.520).The overall positive rate for 13 pathogens detection by LAMP chip was statistically higher than that by bacterial culture(74.39%vs 31.71%,P<0.01).LAMP chip method had the better detection effect on methicillin-resistant Staphylococcus aureus(8.54%vs 0,P<0.05)and common respiratory pathogens,such as Tubercle bacilli,Legionella,Mycoplasma and Chlamydia,which weren't determined by ordinary bacterial culture.Conclusion LAMP chip can quickly detect common respiratory pathogens in BALF of HAP patients,which is helpful for the early identification of HAP pathogens and provides rapid and reliable laboratory evidence for targeted treatment.