| 弓形虫PCR检测试剂盒的组装和初步应用 |
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| 引用本文： | 任科研,李琳,苑淑贤,尹荣兰,杨金生,姚新华.弓形虫PCR检测试剂盒的组装和初步应用[J].中国畜牧兽医,2012,39(9):92-94. |
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| 作者姓名： | 任科研 李琳 苑淑贤 尹荣兰 杨金生 姚新华 |
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| 作者单位： | 吉林省畜牧兽医科学研究院,吉林长春 130062 |
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| 基金项目： | 吉林省科技发展计划重点项目 |
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| 摘 要： | 研制弓形虫PCR检测试剂盒,观察临床应用效果。根据GenBank公布的弓形虫MIC3基因序列,设计合成1对特异性引物,对PCR反应条件进行优化,分别对试剂盒的特异性、敏感性、重复性、稳定性和保存期进行研究,之后进行临床试验。该诊断试剂盒能扩增出弓形虫特异性基因MIC3片段,与猪球虫、贾第虫、猪旋毛虫和隐孢子虫无交叉反应;该检测方法最低能够检测到10个弓形虫速殖子的DNA,不同批次的试剂盒对同一样品检测和同一批次对不同样品检测,有较高的重复性和稳定性,且常温下可保存1年以上,临床应用效果显著。该检测试剂盒对弓形虫病早期诊断、流行病学调查和卫生检疫提供了检测手段。
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| 关 键 词： | 弓形虫 PCR检测试剂盒 组装 应用 |
| 收稿时间： | 2012-02-09 |
Development and Preliminary Application of PCR Detection Kit for Toxoplasma |
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| REN Ke-yan
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LI Lin
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YUAN Shu-xian
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YIN Rong-lan
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YANG Jin-sheng
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YAO Xin-hua.Development and Preliminary Application of PCR Detection Kit for Toxoplasma[J].China Animal Husbandry & Veterinary Medicine,2012,39(9):92-94. |
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| Authors: | REN Ke-yan LI Lin YUAN Shu-xian YIN Rong-lan YANG Jin-sheng YAO Xin-hua |
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| Institution: | Animal Husbandry and Veterinary Science Research Institute of Jilin Province, Changchun 130062,China |
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| Abstract: | To develop PCR detection kit for Toxoplasma and observe the effects of clinical applicaytion in this study.The MIC3 gene sequences of Toxoplasma were chosen to design the species-specific primers on GenBank.The PCR amplification conditions were optimized and a series of tests were conducted respectively in the specificity,sensitivity,stability,reproducibility,and stored period of the kit,and expanded clinical trials were also carried out.This kit could specifically amplify MIC3 gene the expected DNA fragment of Toxoplasma.There was no cross-reaction with other parasites,such as Coccidian,Giardia,Trichinella and Cryptosporidium.It was able to detect as low as 10 of Toxoplasma gondii tachyzoites DNA.The results of different batches of kits of the same sample were consistent,and had higher repeatability and stability.The kit worked well after being stored at room temperature for at least twelve months.Its effect of clinical application was significant.The results turned out that this PCR detection kit could provide a means of early diagnosis,epidemiological investigation and quarantine of Toxoplasma. |
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| Keywords: | Toxoplasma PCR detection kit development application |
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