中国动物检疫
中國動物檢疫
중국동물검역
CHINA ANMAL QUARANTINE
2014年
8期
70-72,85
,共4页
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
何世成%林源%王昌建%劉道新%譚丹%唐小明%王衛國%魯杏華%邱立新%何玉玲
하세성%림원%왕창건%류도신%담단%당소명%왕위국%로행화%구립신%하옥령
RNA提取%PRRSV%TRIZOL%吸附柱%磁珠法
RNA提取%PRRSV%TRIZOL%吸附柱%磁珠法
RNA제취%PRRSV%TRIZOL%흡부주%자주법
RNA extraction%PRRSV%Trizol%adsorption column%magnetic bead extraction
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。
目的：研究Trizol RNA提取法、杭州博日吸附柱RNA提取法、Promega覈痠提取儀磁珠RNA提取法、西安天隆覈痠提取儀磁珠RNA提取法、Thermo覈痠提取儀磁珠RNA提取法等五種不同覈痠提取方法對不同含量豬繁殖與呼吸綜閤徵病毒（PRRSV）暘性樣品的相對提取效率，為動物疫病鑑測診斷實驗室覈痠提取方式的選擇提供技術參攷。方法用五種不同覈痠提取方法對4份10倍梯度稀釋的PRRSV暘性血清、2份PRRSV暘性組織懸液及其離心後的上清液共8份樣品同時進行覈痠提取，用熒光RT-PCR方法進行檢測，穫取各樣品PRRSV熒光RT-PCR結果的CP值，分析五種提取方式的相對提取效率。結果對閤適的提取樣品，磁珠提取法相對提取效率最高，吸附柱法其次，Trizol提取法相對效率較差；在三種磁珠提取法中，提取效率相差在2箇CP值內，差異不明顯，但以天隆磁珠提取法提取效率更高。結論預裝試劑結閤儀器自動化操作的磁珠法提取效果穩定，效率最高；國產儀器國產試劑提取效率不亞于進口儀器和試劑。
목적：연구Trizol RNA제취법、항주박일흡부주RNA제취법、Promega핵산제취의자주RNA제취법、서안천륭핵산제취의자주RNA제취법、Thermo핵산제취의자주RNA제취법등오충불동핵산제취방법대불동함량저번식여호흡종합정병독（PRRSV）양성양품적상대제취효솔，위동물역병감측진단실험실핵산제취방식적선택제공기술삼고。방법용오충불동핵산제취방법대4빈10배제도희석적PRRSV양성혈청、2빈PRRSV양성조직현액급기리심후적상청액공8빈양품동시진행핵산제취，용형광RT-PCR방법진행검측，획취각양품PRRSV형광RT-PCR결과적CP치，분석오충제취방식적상대제취효솔。결과대합괄적제취양품，자주제취법상대제취효솔최고，흡부주법기차，Trizol제취법상대효솔교차；재삼충자주제취법중，제취효솔상차재2개CP치내，차이불명현，단이천륭자주제취법제취효솔경고。결론예장시제결합의기자동화조작적자주법제취효과은정，효솔최고；국산의기국산시제제취효솔불아우진구의기화시제。
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.
