Establishment And Application Of SYBR Green QPCR Assayfor M.Suis Detection

Objective:Establish a method to detect Mycomplasma suis (M.suis).Method:The standard recombinant plasmid was prepared using the PCR production of 16S rRNA gene of M.suis. Then the standard curve was successful to establishment using the plasmid. Specificity and reproducibility test of qPCR was detected by sensitivity test.Results:A 157 bp DNA fragment of the 16S rRNA gene from M.suis was amplified by qPCR. No PCR products were amplified from the samples of PRRSV, PCV2, PPY, PRV, E.coli and S.aureus. The Linearly dependent coefficient and Amplification efficiency of standard curve for qPCR was 0.996 and 98.4% rsepectively. The sensitivity of qPCR was 100 times to that of conventional PCR. The difference in detecting blood sample from swine by qPCR and conventional PCR was not significant(p>0.05).Conclution:The SYBR Green qPCR method had been developed for the detection of M.suis infection successfully. It had high Sensitivity, good Specificity and repeatability, and was suitable for quantitative detection of M.suis in clinical and laboratory.