Establishment of method to detect legionella pneumophila by LAMP technology / 国际检验医学杂志

ABSTRACT

Objective To establish a accurate and rapid method of loop-mediated isothermal amplification(LAMP) for detecting Legionella Pneumophila(LP) .Methods The strains of LP ,Staphylococcus aureus ,Pseudomonas aeruginosa ,Escherichia coli ,Enterobacter sakazakii ,Listeria monocytogenes ,Salmonella typhimurium ,shigella flexneri and Vibrio parahaemolyticus were selected . According to six special domains of toxicity-related mip gene on LP ,the LAMP primers(mip-1 ,mip-2 ,mip-3) were designed by using the Primer Explorer Version 4 .0 .The genomic DNA was extracted for conducting LAMP .Then its specificity ,lowest detectable limit and stability were evaluated .Results The screened primer mip-3 appeared the peak after amplification for about 10 min in the LAMP reaction system for detecting LP ,moreover the peak value was higher ;while the strains of non-LP had no amplification reaction ;the LAMP detection limit could reach 100 fg/μL .The primer mip-3 appeared the peak almost at the same time in 20 times of duplicated detection ,and its stability was good .Conclusion The established LAMP detection method has the advantages of strong specificity and high stability ,can rapidly and accurately detect LP and has large prospect of promotion and application .