Establishment and Application of Methods for Detecting H5N1 Pathogen with Multiple Probe RT-PCR / 现代检验医学杂志

ABSTRACT

Objective To establish and apply the multiple probe RT-PCR analysis for H5N1 pathogen detection methods. Methods Used the gene sequences published in GenBank,which were designed and synthesized two pairs of specific primers (P1/P2,P3/P4)accorded the conserved regions.Through the optimization of amplification conditions to establish the rapid detection of two viruses duplex PCR method.Results The suitable primer concentrationP1/P2 was 0.32μmol/L,P3/P4 was 0.96μmol/L.The amount of the minimum virus nucleic acid was 2 ng/μl by the double detect,H5N1 pathogen mixed were occurred specific bands in the corresponding location,and other viruses and blank control strips had not bands.Conclu-sion A variety of probes for RT-PCR analysis H5N1 pathogen detection methods in a reasonable application of primer con-centration under has good sensitivity and specificity,and it is worth the inspection application.