| 猪链球菌2型分泌核酸酶(SsnA)的表达、ELISA方法建立及其在鉴别诊断中的应用 |
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| 引用本文： | 朱剑,康超,张安定,金梅林.猪链球菌2型分泌核酸酶(SsnA)的表达、ELISA方法建立及其在鉴别诊断中的应用[J].畜牧兽医学报,2011,42(2). |
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| 作者姓名： | 朱剑 康超 张安定 金梅林 |
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| 作者单位： | 华中农业大学,农业微生物学国家重点实验室,武汉,430070 |
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| 基金项目： | 国家973计划(2010CB534000); 国家科技重大专项课题(2008ZX10004-013;2009ZX10602-14); 转基因生物新品种培育重大专项(2009ZX08009-141B); 国家公益性行业农业专业专项资助(200803018); 湖北省自然科学基金(2009CDB171) |
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| 摘 要： | 猪链球菌2型(Streptococcussuis type 2,SS2)是一种广泛分布的人畜共患病原菌,严重危害着人类健康和养猪业的发展。本试验旨在研究分泌核酸酶基因(SsnA)作为分子诊断标识在对SS2感染的鉴别诊断中所起的作用。作者以SS2-LT菌株基因组DNA为模板,通过PCR扩增SsnA基因片段,将该片段克隆到表达载体pET-28a(+)中,在大肠杆菌BL21(DE3)中成功表达,获得大小为46 ku的融合蛋白,经SDS-PAGE和Western blot分析表明,重组蛋白质具有抗原反应活性。以纯化的融合蛋白作为包被抗原,建立了SsnA-ELISA鉴别诊断方法。对已知背景的人工感染及疫苗免疫血清和临床血清进行检测,比较感染血清和免疫血清的检测结果,结果发现两者差异显著(P<0.05),表明建立的SsnA-ELISA方法具有良好的检测效果。利用该方法对采集自8个省份的1 446份血清进行检测,显示SS2的阳性率为0%~14.77%,平均阳性率为5.39%。本研究结果表明:猪链球菌2型SsnA可以作为一个良好的鉴别感染的分子诊断标识;对临床血清的检测结果表明感染及流行与气候环境存在明显的关系。
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| 关 键 词： | 猪链球菌2型 分泌核酸酶(SsnA) 鉴别诊断 ELISA |
Expression of Streptococcus suis Type 2 Secreted Nuclease A (SsnA),Development of the SsnA-ELISA and Its Application in the Differential Diagnosis |
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| ZHU Jian,KANG Chao,ZHANG An-ding,JIN Mei-lin.Expression of Streptococcus suis Type 2 Secreted Nuclease A (SsnA),Development of the SsnA-ELISA and Its Application in the Differential Diagnosis[J].Acta Veterinaria et Zootechnica Sinica,2011,42(2). |
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| Authors: | ZHU Jian KANG Chao ZHANG An-ding JIN Mei-lin |
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| Institution: | ZHU Jian,KANG Chao,ZHANG An-ding,JIN Mei-lin (State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070,China) |
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| Abstract: | Streptococcus suis serotype 2(SS2) is a zoonotic infectious pathogen associated with a wide range of diseases in human health and pig industry.This article aims to study the role of SsnA gene as a molecular diagnosis marker in the differential diagnosis of SS2.SsnA gene was amplified using SS2-LT genome DNA as template and the sequence was then inserted into expression vector pET-28a(+).The plasmid was introduced into E.coli BL21(DE3) for expression and 46 kD recombinant protein was obtained,which had been ... |
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| Keywords: | Streptococcus suis serotype 2 secreted nuclease A(SsnA) differential diagnosis ELISA |
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