| 王花茹,王长军,陆承平,潘秀珍,陶开华,唐家琪.致病性猪链球菌主要毒力因子基因多重PCR检测[J].Chinese journal of Epidemiology,2005,26(9):640-644 |
| 致病性猪链球菌主要毒力因子基因多重PCR检测 |
| Detection of virulence-associated factors of Streptococcus suis by multiplex PCR assay |
| Received:June 30, 2005 |
| DOI： |
| KeyWord: 猪链球菌2型 毒力因子 荚膜多糖 溶菌酶释放蛋白 胞外因子 溶血素 |
| English Key Word: Streptococcus suis serotype 2 Virulence-associated factors Capsular polysaccharide Muraminidase released protein Extracellular factor Suilysin |
| FundProject:“十五”国家重要传染病科技攻关计划资助项目(2003BA712A03-05) |
| Author Name | Affiliation | E-mail | | WANG Hua- ru | Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002,China | | | WANG Chang-jun | Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002,China | | | LU Cheng-ping | Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002,China | | | PAN Xiu-zhen | Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002,China | | | TAO Kai- hua | Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002,China | | | TANG Jia-qi | Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002,China | tjq85@hotmail.com |
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| Abstract: |
| 目的 建立多重PCR体系,实现猪链球菌毒力相关因子基因cps、mrp、epf(epf*)和sly的同步检测。方法 根据上述毒力因子基因核苷酸序列,设计和合成4对特异性引物,通过体系和条件优化,建立多重PCR检测方法,并对72株种属背景明确的实验菌株(其中猪链球菌48株、阴性对照株24株)及49株猪临床分离的链球菌样本进行检测分析。结果 48株猪链球菌中,cps2检出率为33.3%(16/48)、mrp检出率为29.2%(14/48)、epf检出率为25%(12/48)、epf*检出率为6.3%(3/48)、sly检出率为(54.2%)(26/48), 上述检测结果与菌株的毒力因子背景情况完全相符;24株阴性对照和49株猪临床分离样本毒力因子检测结果均为阴性。结论 多重PCR可用于猪链球菌毒力相关因子CPS2、MRP、EF、Sly的基因检测, 特异性和敏感性好, 为该病快速诊断和分子流行病学调查提供了新的技术手段。54.2%(26/48),上述检测结果与菌株的毒力因子背景情况完全相符;24株阴性对照和49株猪临床分离样本毒力因子检测结果均为阴性。结论 多重PCR可用于猪链球菌毒力相关因子CPS2、MRP、EF、Sly的基因检测,特异性和敏感性好,为该病快速诊断和分子流行病学调查提供了新的技术手段。 |
| English Abstract: |
| Objective To rapidly and sensitively detect the four virulence-associated factors of Streptococcus suis,a multiplex PCR was developed. Methods In the process of this reactiont four distinct DNA targets were amplified. One target was based on the serotype 2(and 1/2) specific cps gene and the others were based on Streptococcus suis mrp, epf (epf*) and sly gene, encoding the MRP ≥EF(EF*) and Sly proteins of Streptococcus suis. 72 isolates, which including 48 strains of Streptococcus suis and 24 strains of negative control,and 49 clinical specimens were detected by the multiplex PCR assay. Results All PCR products were detected by electrophoresis on 1.2% agarose gels. With the 48 Streptococcus suis strains, the positive detection rates of cps2+, mrp≥, epf +,epf* + and sly+ were 16/48,14/48,12/48,3/48 and 26/48,respectively. The results were confirmed by bacteriological examination. There were no specific amplification products including 49 clinical specimens and 24 negative control strains. Conclusion The results demonstrated that multiplex PCR was a highly specific and sensitive diagnostic tool for the detection of virulence-associated factors of streptococcus suis. |
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