Loop-mediated Isothermal Amplification For Detection Of Lower Respiratory Tract Infection Pathogenic Bacterium

Objective To study the clinical application of LAMP for common lower respiratory tract infection pathogenic bacterium detection,improve the speed and accuracy of bacteria detection. To confirm it utilization value in diagnosis lower respiratory tract infection.Methods Definited 11 species of common lower respiratory tract infection pathogenic bacterium. According to the analysis of lower respiratory tract infection pathogenic bacterium conservative genes,we designed a pair of every bacteria's LAMP primer sequence use BLAST,Align X and Primer5.0 program.Collecting sputums of lower respiratory tract infections.DNA are isolated from each pathogen,and use LAMP to amplify target fragments.Put these reagent on RT-PCR instrument reacting 45 minutes.Determining melting curve.Analysis of the data.The sensitivity and specificity of the detection system was detected.Total 50 samples,which were collected by the routine method of lower respiratory tract infection, were tested by this methods.And compared with tradition sputum germiculture.Results The sensibility of all bacteria achieve to 200copies/25ul reaction system( equivalently 104 copies/ml sputum).Every kind of pathogens only react to its corresponding probes,without consensual reaction of 11 species of bacteria,which show high specificify.LAMP's masccline rate is 82.0%,higher than germiculture. LAMP can detect pathogenic bacterium of low level sample and same hard detect bacteria.Conclusion The method can detect majority pathogens in lower respiratory tract infection successfully,it is sensitive,rapid and specific.In addition,it lay the foudation for lower respiratory tract infection LAMP gene chip researth.