CAJ | 학술논문

以无乳链球菌纤连蛋白fbs基因为主要研究对象,采取环介导等温扩增技术(Loop-Mediated Isothermal Amplification,LAMP),针对fbs基因的6个区域设计4条特异性引物,利用一种链置换DNA聚合酶(Bst DNA polymerase)在63℃保温1 h,通过荧光显色即可完成对无乳链球菌的检测工作.结果显示,LAMP方法能够特异性检测fbs基因,其检测灵敏度是常规PCR方法的100倍,并与实时荧光定量PCR方法相当.所建立的针对无乳链球菌fbs基因的LAMP检测方法具有高度的特异性及稳定性,结果可靠,非常适合无乳链球菌的快速检测.
이무유련구균섬련단백fbs기인위주요연구대상,채취배개도등온확증기술(Loop-Mediated Isothermal Amplification,LAMP),침대fbs기인적6개구역설계4조특이성인물,이용일충련치환DNA취합매(Bst DNA polymerase)재63℃보온1 h,통과형광현색즉가완성대무유련구균적검측공작.결과현시,LAMP방법능구특이성검측fbs기인,기검측령민도시상규PCR방법적100배,병여실시형광정량PCR방법상당.소건립적침대무유련구균fbs기인적LAMP검측방법구유고도적특이성급은정성,결과가고,비상괄합무유련구균적쾌속검측.
To develop a technique for detecting the fbsB gene of Streptococcus agalactiae using a novel DNA amplification method designated Loop-Mediated Isothermal Amplification (LAMP).The detection assay is based on the loop-mediated isothermal amplification (LAMP) reaction.The fbsB gene was amplified by a set of four specially primers that recognize six distinct sequences of the target.The amplification can be obtained in 1 h by incubating all of the reagents in a single tube with Bst DNA polymerase at 63℃.Results showed that the developed LAMP assay demonstrated an exceptionally higher than the conventional PCR.This assay results was found to be 100 times more sensitive than the PCR assay,and consistent with the results of real-time PCR.Our results clearly demonstrate that the LAMP-based assay is a sensitive and reliable means for the detection of Streptococcus agalactiae.