环介导恒温扩增法检测痰液标本中肺炎链球菌

The detection of Streptococcus pneumoniae in sputum samples by loop-mediated isothermal amplification method

目的建立环介导恒温扩增(LAMP)技术快速检测痰标本肺炎链球菌的方法。方法根据基因序列数据库(GenBank)的肺炎链球菌R6株lytA基因序列(AE008540)设计特异LAMP引物,对痰液标本进行LAMP法检测,同时进行传统微生物检测和PCR检测。结果传统微生物检测肺炎链球菌阳性的27例标本,LAMP法检测和PCR检测也为阳性;在传统微生物检测肺炎链球菌阴性的27例标本中,LAMP法有9例检出阳性,PCR法有4例阳性,统计分析表明三种方法的阳性率无显著性差异。结论三种方法检测的阳性率无显著性差异,但LAMP法能够更快速、简便地检出肺炎链球菌感染。

Objective To establish a method of loop-mediated isothermal amplification for rapidly detecting Streptococcus pneumoniae in sputum samples.Methods Four S.pneumoniae-specific LAMP primers which be aimed at the gene of autolysin(lytA) were designed from the published sequence of strain R6,then detected Streptococcus pneumoniae in sputum samples by LAMP method and detected that by traditional and PCR methods at the same time.Results 54 samples were detected in our experiment and the traditional,PCR and LAMP methods have 27,31 and 36 positive test results respectively.Conclusion In spite of the three methods in our experiment have no significant differences,the LAMP technique is more rapid and convenient compared with PCR method and traditional culture method,so this method is still very useful for the rapid detection of S.pneumoniae.