猪链球菌2型实时荧光定量PCR检测方法的建立与应用

doi:10.16431/j.cnki.1671-7236.2016.12.004

Abstract

Abstract:

In this study,a quantitative Real-time PCR method using the specific primers according to CPS2J gene was established to detect Streptococcus suis serotype 2. The result showed that the equation of standard curve was y=-3.073x+36.87,r=0.995,which demonstrated that the assay had good linear relationship.The melting curve analysis showed that there was only specific peak.Sensitivity test showed that the method could detect the template at the lowest concentration of 1.0×10¹ copies/μL,which was 10 times higher than the ordinary PCR.The specific tests showed that this method could able to detect Streptococcus suis serotype 2 specially and had no cross-reaction with other serotypes or other bacteria from swine. The CV of repeatability test was 0.37% to 0.63%,lower than 2.5%. The clinical diagnosis showed this assay was more sensitive than ordinary PCR and bacteria isolation. All the results showed that the established method was sensitive,specific and reproducible,which could be used for the rapid diagnosis and quantitative detection of Streptococcus suis serotype 2.

Key words: Streptococcus suis serotype 2; quantitative Real-time PCR; sensibility; specificity; repeatability

CLC Number:

S858.28

LI Jian-da, YU Jiang, ZHANG Yu-yu, REN Su-fang, CHEN Lei, GUO Li-hui, SUN Wen-bo, CHEN Zhi, WANG Song, LIU Jun-zhen, DU Yi-jun, LI Jun, YANG Ling-zhi, WANG Jin-bao, WU Jia-qiang. Establishment and Application of Quantitative Real-time PCR Method to Detect Streptococcus suis Serotype 2[J]. , 2016, 43(12): 3107-3113.

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Establishment and Application of Quantitative Real-time PCR Method to Detect Streptococcus suis Serotype 2

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