Loop-mediated isothermal amplification for detecting Streptococcus pneumoniae and its clinical appfication / 中华检验医学杂志

ABSTRACT

Objective To establish loop-mediated isothermal amplification(LAMP)method for detecting Streptococcus pneumoniae in clinical samples.Methods Four Streptococcus pneumoniae-specific LAMP primers were designed according to the published sequence of strain R6(GenBank accession number AE008540).Genomie DNA in positive blood cultures was extracted by nanidine hydrochloride and benzene-methanol.Then lytA was amplified by LAMP at 63℃ for 45 minutes.We observed the turbidity in thereaction tube.For further confirmation.The amplified products were also detected using electrophoresis in 2% agarose gels,followed by ethidium bromide staining.Resuits 16 strains of Streptococcus pneumoniae were detected in 196 positive blood cuhure bottles by LAMP.Compared with traditional method.It8 sensitivity and specificity were both 100%and the detection could be finished in an hour.The assay had a minimum detection limit of 102 CFU/m1.Conclusions This IJAMP-based assay is simple,rapid.Sensitive and specific.It can be used to detect trept OCOCCUS pneumoniae in clinical samples.