利用环介导恒温扩增16S rRNA对无乳链球菌快速鉴定的新方法

Rapid detection of Group B Streptococcus specific gene 16S rRNA by loop-mediated isothermal amplification

目的建立用环介导恒温扩增16S rRNA对无乳链球菌鉴定的方法。方法 52株无乳链球菌和131株非无乳链球菌为研究对象,在无乳链球菌16S rRNA处设计4条特异性引物,对环介导恒温扩增的条件优化后,验证其特异性和敏感性。结果 52株无乳链球菌均能有效检出,没有任何非无乳链球菌的检出;环介导恒温扩增的最低检出限为40CFU/ml,比普通的PCR高出100倍。结论环介导恒温扩增16S rRNA,能方便、敏感、特异地完成对无乳链球菌的快速鉴定,适于基层机构使用。

Objective To develop a new method to rapidly identify Streptococcus agalactiae by loop-mediated isothermal amplificating 16S rRNA.Methods 52 Streptococcus agalactiae isolates and 131 non-Streptococcus agalactiae isolates were used as the research objects.Four sets of primers were designed to recognized Streptococcus agalactiae specific gene 16S rRNA.After a optimal reaction condition was selected,its specificity and sensitivity were evaluated by LAMP reaction.Results 52 Streptococcus agalactiae isolates were all positive,and the non-Streptococcus agalactiae isolates were negative.The results showed that the LAMP,with a detection limit of 40CFU/ml,was more sensitive by 100 times than the ordinary PCR.Conclusions The LAMP method for detection of Group B Streptococcus specific gene 16S rRNA is a potential and valuable method due to its rapidity,simplicity,and accuracy.It is especially suitable for grass-roots medical units.