| 五种核酸提取法对猪繁殖与呼吸综合征病毒提取效率的比较 |
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| 引用本文： | 何世成,林源,王昌建,刘道新,谭丹,唐小明,王卫国,鲁杏华,邱立新,何玉玲.五种核酸提取法对猪繁殖与呼吸综合征病毒提取效率的比较[J].中国动物检疫,2014,31(8):70-72. |
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| 作者姓名： | 何世成 林源 王昌建 刘道新 谭丹 唐小明 王卫国 鲁杏华 邱立新 何玉玲 |
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| 作者单位： | [1]湖南省动物疫病预防控制中心,湖南长沙410014; [2]长沙市动物疫病预防控制中心,湖南长沙410006 |
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| 摘 要： | 目的：研究Trizol RNA提取法、杭州博日吸附柱RNA提取法、Promega核酸提取仪磁珠RNA提取法、西安天隆核酸提取仪磁珠RNA提取法、Thermo核酸提取仪磁珠RNA提取法等五种不同核酸提取方法对不同含量猪繁殖与呼吸综合征病毒（PRRSV）阳性样品的相对提取效率，为动物疫病监测诊断实验室核酸提取方式的选择提供技术参考。方法用五种不同核酸提取方法对4份10倍梯度稀释的PRRSV阳性血清、2份PRRSV阳性组织悬液及其离心后的上清液共8份样品同时进行核酸提取，用荧光RT-PCR方法进行检测，获取各样品PRRSV荧光RT-PCR结果的CP值，分析五种提取方式的相对提取效率。结果对合适的提取样品，磁珠提取法相对提取效率最高，吸附柱法其次，Trizol提取法相对效率较差；在三种磁珠提取法中，提取效率相差在2个CP值内，差异不明显，但以天隆磁珠提取法提取效率更高。结论预装试剂结合仪器自动化操作的磁珠法提取效果稳定，效率最高；国产仪器国产试剂提取效率不亚于进口仪器和试剂。
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| 关 键 词： | RNA提取 PRRSV TRIZOL 吸附柱 磁珠法 |
Comparison of ifve RNA extraction methods for extraction of PRRS virus nucleic acids |
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| Institution: | He Shicheng,Lin Yuan,Wang Changjian,Liu Daoxin,Tan Dang,Tang Xiaoming,Wang Weiguo,Lu Xinhua,Qiu Lixin,He Yuling ( 1. Hunan Center for Animal Disease Prevention and Control, Changsha, Hunan 410014; 2. Changsha Center for Animal Disease Prevention and Control, Changsha, Hunan 410006) |
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| Abstract: | Aim To compare the extraction efifciency of ifve RNA extraction methods（Trizol RNA extraction method、adsorption column RNA extraction of BIOER,magnetic bead RNA extraction of Promega,XianTianlong and Ther-mo）for detection of PRRSV viruses,so as to provide technical reference for animal disease diagnostic laboratories in nucleic acid extraction. Methods PRRSV RNAs were extracted using the 5 RNA extraction methods on positive serum samples of PRRS viruses diluted serially from 10-1 to 10-4 and 2 PRRSV positive tissue suspension and supernatant at the same time. The RNA preparations were quantiifed by fluorescence real-time quantitative RT-PCR. Results For the ap-propriate samples,the extraction efifciency of magnetic bead was the highest;the adsorption column RNA extraction followed;and Trizol RNA extraction method was lower;there was 2 CP value deference among the three magnetic bead RNA extraction methods,indicating no obvious difference,but the Tianlong magnetic bead extraction efifciency was higher relatively. Conclusion Pre-installed reagent combination with automation magnetic beads to extract was stable and gave the highest efifciency. The magnetic bead extraction efifciency had no signiifcant difference between the domestic and imported products. |
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| Keywords: | RNA extraction PRRSV Trizol adsorption column magnetic bead extraction |
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