呼吸道病原体核酸恒温扩增芯片十三联检在下呼吸道感染常见病原体基因中的检测
The Application of Loop-mediated Amplification Chip in Gene Testing of Common Lower Respiratory Pathogens
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摘要: 目的 探讨呼吸道病原体核酸环介导恒温扩增芯片法 (Loop-mediated amplification, LAMP) 十三联检在下呼吸道感染常见病原体基因诊断中的价值.方法 收集疑似下呼吸道感染患者的深部痰液或肺泡灌洗液样本122例, 分别采用LAMP技术、病原体分离与培养、实时荧光定量PCR (FQ-PCR) 技术检测病原体, 应用SPSS软件及Kappa检验将检测结果按标本类型、年龄段及检测方法进行比对分析.结果 122例呼吸道感染患者标本中, 痰液病原体阳性率 (55.56%) 高于肺泡灌洗液阳性率 (46.88%) , 病原体种类分别以耐甲氧西林葡萄球菌 (MASA) 和肺炎支原体 (MP) 为主 (P<0.05) ;高年龄段 (≥60岁) 以检出MASA为主, 而低年龄段 (≤20岁) 以检出MP为主 (P<0.01) ;LAMP十三联检阳性检出率 (50.00%) 高于病原体分离培养的阳性检出率 (36.67%) , 2种检测方法对肺炎克雷伯菌 (Kpn) 和鲍曼不动杆菌 (CSAB) 的检出率具有一致性 (Kappa>0.4) , 而其它病原体阳性结果的一致性较低 (Kappa<0.4) ;LAMP与实时FQ-PCR法对MP的检测结果具有较高的一致性 (Kappa>0.7) .结论 呼吸道病原体LAMP十三联检具有快速、灵敏、高效、特异及病原体覆盖面广的优势, 可对临床提供快速可靠的实验室诊断依据.呼吸道标本类型及检验方法不同, 其病原体检验结果也可能有所不同, 各具优缺点, 需要临床医生给予密切关注.Abstract: Objective To investigate the value of Loop-mediated Amplification (LAMP) in the gene diagnosis of common pathogens in lower respiratory infection.Methods One hundred and twenty-two cases of lower respiratory tract secretions and bronchoalveolar lavage fluid were collected from patients with suspected lower respiratory infection.The pathogens were tested by using LAMP, isolation and cultivation of microbial pathogens, real-time FQ-PCR.The results were compared with different specimen types, age and detection methods.Statistical analysis was performed using the SPSS software and Kappa test.Results In 122 cases of specimensfrom patients with respiratory infection, the positive rate of sputum was higher (55.56%) than that in bronchoalveolar lavage fluid (46.88%) .The main pathogens were Methicillin-resistant Staphylococcus (MASA) and mycoplasma pneumoniae (MP) (P<0.05) respectively.In different age stage, MASA was mainly detected in the elder group (>60 years) , and MP was the main pathogen in the younger patients (≤20 years) (P<0.01) .The detection rate of LAMP (50.00%) was higher than the microbial Isolation and cultivation (36.67%) .The detection rates of Klebsiella pneumoniae (Kpn) and Bauman Acinetobacter (CSAB) by two different methods were consistent (kappa>0.4) , however the consistencies in other pathogens were low (kappa<0.4) .And there was a high consistency between LAMP and real-time FQ-PCR in the detection of MP (kappa >0.7) .Conclusions LAMP has the advantages of rapidity, sensitivity, high effciency and specificity, high throughput and wide coverage of pathogens, which may be a quick and reliable laboratory diagnosis method.And using different respiratory specimens or methods may have different results.They have their own advantages and disadvantages, which are needed to be paid close attention by clinicians.
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Key words:
- Lower respiratory infection /
- Pathogen /
- Loop-mediated amplification
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