| 破碎法提取家蚕微孢子虫孢子核酸的PCR检测灵敏度试验 |
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| 引用本文： | 李光才,蔡顺风,何祥康,何欣怡,马焕艳,孟祥坤,邱海洪,何永强,鲁兴萌. 破碎法提取家蚕微孢子虫孢子核酸的PCR检测灵敏度试验[J]. 蚕桑通报, 2011, 42(4): 8-11 |
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| 作者姓名： | 李光才 蔡顺风 何祥康 何欣怡 马焕艳 孟祥坤 邱海洪 何永强 鲁兴萌 |
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| 作者单位： | 1. 浙江大学蚕蜂研究所,浙江杭州,310058
2. 浙江出入境检验检疫局技术中心,浙江杭州,310012 |
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| 基金项目： | 现代农业产业技术体系建设专项，浙江省科技厅重大项目，国家质量监督检验检疫总局科研项目 |
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| 摘 要： | 蚕微粒子病检测技术是该病防治中的一项重要技术，本文在通过破碎法提高模板核酸获取量的基础上，对纯化家蚕微粒子虫孢子的PCR检测灵敏度进行了测试。提取核酸-模板稀释测试12对引物的灵敏度中有4对引物达到0．05粒（10^2粒/mL）、4对引物5粒（10^4粒/mL）、1对引物50粒（10^5粒/mL）、2对引物500粒（1...
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| 关 键 词： | 家蚕微粒子病 孢子 聚合酶链式反应 检测 灵敏度 |
Study on the Detection Sensitivity of PCR Method for Silkworm Pebrine Diseases |
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| LI Guang-cai,CAI Shun-feng,HE Xiang-kang,HE Xin-yi,MA Huan-yan,MENG Xiang-kun,QIU Hai-hong,HE Yong-xiang,LU Xing-meng. Study on the Detection Sensitivity of PCR Method for Silkworm Pebrine Diseases[J]. Bulletin of Sericulture, 2011, 42(4): 8-11 |
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| Authors: | LI Guang-cai CAI Shun-feng HE Xiang-kang HE Xin-yi MA Huan-yan MENG Xiang-kun QIU Hai-hong HE Yong-xiang LU Xing-meng |
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| Affiliation: | 1(1.Institute of Sericulture and Apiculture,Zhejiang University,Hangzhou Zhejiang 310029,China;2.Technical Center of Zhejiang Entry-Exit Inspection & Quarantine Bureau,Hangzhou Zhejiang 310012,China) |
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| Abstract: | Silkworm pebrine disease detection technology is an important technique in the prevention and treatment for pebrine. Based on the higher DNA recovery by acid-washed glass beads treatment, detection sensitivity of PCR method for purified Nosema bombycis spores was tested. In DNA extraction - template dilution test, sensitivity of 12 pairs of primers are as follows： 4 pairs of primers are 0.05 spore （10^2 spore/mL）, 4 pairs of primers are 5 spores （10^4 spore/mL）, 1 pair of primers is 50 spores （10^5 spore/ mL）, 2 pairs of primers are 500 spores （10^6 spore/mL）, 1 pair of primers is 5000 spores （10^7 spore/mL）. The repeatability of sensitivity thresholds of NbZSO04F/NbZS004R （0.05） and NbZS007F/NbZS007R （0.005） was 90%. In spore dilution - DNA extraction test, detection sensitivity of NbZS004F/NbZS004R and NbZS007F / NbZS007R are 5 spores （10^4 spore/mL） and 0.5 spores （10^3 spore/mL）, respectively. |
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| Keywords: | Silkworm Pebrine disease Spore PCR Detection Sensitivity |
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