| 猪弓形虫特异PCR诊断方法的建立 |
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| 引用本文： | 张云峰,刘福元,徐雪萍,王光雷.猪弓形虫特异PCR诊断方法的建立[J].新疆农业科学,2010,47(9):1823. |
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| 作者姓名： | 张云峰 刘福元 徐雪萍 王光雷 |
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| 作者单位： | 新疆农垦科学院畜牧兽医研究所,新疆石河子,832000;新疆畜牧科学院兽医研究所,乌鲁木齐,830000 |
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| 基金项目： | 新疆生产建设兵团农业科技攻关计划项目，国家绒毛用羊产业技术体系建设专项资金 |
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| 摘 要： | 目的]建立特异PCR方法诊断猪弓形虫病.方法]以核糖体DNA第一内转录间隔区(ITS1)序列作为弓形虫种特异的遗传标记,优化PCR反应条件,对猪弓形虫分离株和临床病料进行检测.随机取阳性PCR扩增片段进行克隆,测序鉴定.结果]在猪弓形虫分离株中PCR扩增出目的条带,且在肺门淋巴结和肠系膜淋巴结中扩增出特异性条带;PCR方法能检测到的最低DNA量为7.81 pg/μL;测序结果显示核苷酸序列与Genbank中已登录的猪弓形虫IST1基因序列相应部分序列完全相同.结论]该PCR方法具有较高的敏感性和特异性,是诊断猪弓形虫病的一种快速检测方法.
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| 关 键 词： | 猪 弓形虫 PCR 诊断 |
| 收稿时间： | 2010-09-25 |
Establishment of Specific PCR Assay for Diagnosing Swine Toxoplasma gondii |
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| ZHANG Yun-feng,LIU Fu-yuan,XU Xue-ping,WANG Guang-lei.Establishment of Specific PCR Assay for Diagnosing Swine Toxoplasma gondii[J].Xinjiang Agricultural Sciences,2010,47(9):1823. |
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| Authors: | ZHANG Yun-feng LIU Fu-yuan XU Xue-ping WANG Guang-lei |
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| Abstract: | 【Objective】To establish specific PCR assay for diagnosing swine Toxoplasmosis.【Method】The first internal transcribed spacer sequence(IST1) of ribosomal DNA was used as the specific genetic marker for Toxoplasma gondii,the PCR reaction condition was optimized,the isolated strain and clinical infected materials of swine Toxoplasma were tested.Positive PCR amplified fragment was cloned and sequenced.【Result】DNA specific fragment for T.gondii strain was amplified,and specific bands were amplified in the bronchopulmonary lymph node and mesenteric lymph node ;the assay was able to detect as little as 7.81 pg/uL DNA of T.gondii;the nucleotide sequence was completely the same to the published homologous IST1 gene sequence of swine T.gondii in Genbank.【Conclusion】The PCR assay was highly sensitive and specific,which is a rapid method for diagnosing swine toxoplasmosis. |
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| Keywords: | PCR |
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