A Method for Detecting Adhesive Related-Factors of Streptococcus suis Serotype 2 by Real-time PCR

作     者：汪伟 何孔旺 倪艳秀 周俊明 张雪寒 俞正玉 吕立新 茅爱华 温立斌 王小敏 李彬 郭容莉 Wei WANG;Kongwang HE;Yanxiu NI;Junming ZHOU;Xuehan ZHANG;Zhengyu YU;Lixin LU;Aihua MAO;Libin WEN;Xiaomin WANG;Bin LI;Rongli GUO

作者机构：江苏省农业科学院兽医研究所农业部兽用生物制品工程技术重点实验室国家兽用生物制品工程技术研究中心江苏南京210014 

基　　金：Supported by National Natural Science Foundation of China(31072155) Natural Science Foundation of Jiangsu Province(BK2010068) Fund for Independent Innovation of Agricultural Science in Jiangsu Province[CX(11)2060] Special Fund for Agroscientific Research in the Public Interest(201303041) 

出 版 物：《Agricultural Science & Technology》 (农业科学与技术（英文版）)

年 卷 期：2013年第14卷第10期

页      码：1378-1382页

摘      要：[Objective] This study aimed to establish a method for quantitative detection of mRNA transcriptional level of SS2 adhesive related-factors of Streptococcus suis serotype 2 （SS2） by fluorescent quantitative PCR. []Vlethod] The gene fragments en- coding SS2 adhesive related-factors MRP, FBPS and CPS2J and a housekeeping gene aroA were amplified by reverse transcription PCR from the total RNA of SS2, cloned, and sequenced. The recombinant plasmids containing the target genes were constructed, and used as templates in Real-time PCR. [Result] Dynamic curves, stan- dard curves and melting curves of the adhesive related-factors and aroA were ob- tained by the optimized Real-time PCR system. The standard curves showed a good linear relationship between template copy number and circulation number, and the correlation coefficients （FF） of the standard curves were over 0.995. Also, these as- says were highly specific a^d there was single specific melting peak for every gene. Moreover, the assays were highly sensitive and had a detection limit of 1.0×102 copies in 1 μl of initial templates. Finally, it was highly repeatable and had a coeffi- cient of variation less than 2% for intra-assay. [Conclusion] This study will provide a way to reveal the adhesion mechanism of SS2 to different host cells at molecular level.

主 题 词：Streptococcus suis serotype 2 Adhesive related-factors (adhesins) Real- time PCR 

学科分类：1001[医学-基础医学] 10[医学] 

D　O　I：10.16175/j.cnki.1009-4229.2013.10.008

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