环介导等温扩增技术在下呼吸道感染常见病原体检测中的应用  

The application of loop-mediated isothermal amplification for detection of common pathogenic bacteria in lower respiratory tract infections

作　　者：陈愉生[1] 王大璇[2] 李鸿茹[1] 陈炆颖[3] 张祥娥[1] 邵慧[4] 许能銮[1] 林明[1] 王璨 闫胜龙 胡辛兰[1] 高占成[1] 

机构地区：[1]福建医科大学省立临床医学院福建省立医院呼吸科,福州350001 [2]福建医科大学省立临床医学院福建省立医院临床医学院,福州350001 [3]福建医科大学省立临床医学院福建省立医院急诊内科,福州350001 [4]福建医科大学省立临床医学院福建省立医院高于科,福州350001 [5]北京博奥生物有限公司

出　　处：《中华结核和呼吸杂志》2014年第4期270-273,共4页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：福建省科技计划项目（2009Y0010,2012Y0016）;国家“863计划”课题（2006AA02Z4A9）

摘　　要：目的 应用环介导等温扩增（LAMP）技术对下呼吸道感染患者痰标本中常见致病菌进行核酸检测,调查病原菌的分布,研究LAMP技术对下呼吸道感染病原体检测的临床意义.方法 收集福建省立医院呼吸科下呼吸道感染患者289例的合格痰标本289份,LAMP技术检测病原体,分析下呼吸道感染病原菌的分布,并与传统的培养法进行比较.将细菌浓度＞1×103拷贝/ml的病例分为初治已覆盖病原体组和初治未覆盖病原体组,比较两组患者年龄、有无基础疾病、抗感染治疗时间、总住院时间、初治有效率及治愈率.结果 289例患者中培养法阳性44例（15.2％）.LAMP技术检测以细菌浓度＞1 ×103拷贝/ml为界值,判定为阳性结果的有124例（43.0％）.下呼吸道分离的病原菌中细菌144株（77.8％）,非典型病原菌41株（22.2％）;革兰阴性杆菌为主要致病菌,依次为铜绿假单胞菌、流感嗜血杆菌、肺炎克雷伯菌及肺炎链球菌.初治已覆盖病原体组（95例）和初治未覆盖病原体组（29例）的总住院时间分别为（13.9±9.1）和（16.3±11.2）d,差异有统计学意义（t=-2.083,P=0.039）;初治有效率两组分别为31.0％（29/95）和93.1％（27/29）,差异有统计学意义（x2 =31.0,P＜0.01）;两组的抗感染治疗时间[（13.9±9.1）和（16.3±11.2）d]比较,差异无统计学意义（t=-1.073,P=0.285）;两组的治愈率分别为84.2％（80/95）和95.7％（28/29）,差异无统计学意义（x2 =0.6,P=0.4）.结论 LAMP技术简便,可快速扩增肺部感染患者痰液中常见致病菌核酸,检出率明显高于传统的痰培养法.LAMP技术有利于治疗前明确致病菌并进行针对性治疗,缩短患者的住院时间,提高初始治疗的有效率.Objective To investigate the spectrum of common pathogenic bacteria of low respiratory tract infection by loop-mediated isothermal amplification （LAMP） of nucleic acid test and to prove the clinical significance of this method.Methods A total of 289 qualified sputum samples from patients with lower respiratory tract infections in Fujian Province were detected by LAMP technique,and then the distribution of pathogenic bacteria was analyzed.The positive cases （the patients whose specific bacterial copies in their sputum samples ＞ 1 × 103 copies/ml） were divided into 2 groups according to whether their treatment had covered this pathogen or not.The underlying diseases,duration of anti-bacterial treatment,the hospital days,and the effectiveness of initial treatment and cure rate were compared.Results The culture method in the 289 patients showed that 44 （15.2％） were positive.Tests by the LAMP method with a bacteria concentration ＞ 1 × 103 copies/ml as cutoff value,showed positive results in 124 patients （43.0％）.The lower respiratory tract pathogens included 144 strains of bacteria （77.8％）,and 41 strains of atypical pathogens （22.2％）.Gram-negative bacteria were the predominant pathogens,such as Pseudomonas aeruginosa,H.influenzae,Klebsiella pneumoniae,and Streptococcus pneumoniae.In 95 cases the initial therapy had covered the pathogens,while in 29 cases the initial therapy had not.The effectiveness of the initial treatment （x2 =31.0,P ＜ 0.01） and the total days of hospital stay （t =-2.083,P =0.039）in the group whose antibiotics had covered the pathogens were significantly higher than those of the other group.However,there were no significant difference in duration of anti-bacterial treatment （t =-1.073,P =0.285） and cure rates （x2 =0.6,P =0.4） between the 2 groups.Conclusions LAMP method can detect the nuclear acid of the bacteria in the sputum much more rapidly and sensitively than the routine culture method.LAMP technique may be helpful to know the pathogenic b

关 键 词：呼吸道感染 细菌 环介导等温扩增技术 

分 类 号：R446.5[医药卫生—诊断学]