中华实验和临床病毒学杂志
中華實驗和臨床病毒學雜誌
중화실험화림상병독학잡지
CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL VIROLOGY
2010年
2期
147-149
,共3页
王小波%何金生%付远辉%郑娴娴%方璇
王小波%何金生%付遠輝%鄭嫻嫻%方璇
왕소파%하금생%부원휘%정한한%방선
呼吸道合胞病毒,人%滴定分析法%聚合酶链反应%免疫酶试验%半数抑制浓度
呼吸道閤胞病毒,人%滴定分析法%聚閤酶鏈反應%免疫酶試驗%半數抑製濃度
호흡도합포병독,인%적정분석법%취합매련반응%면역매시험%반수억제농도
Respiratory syncytial virus,human%Titrimetry%Polymerase chain reaction%Immunoenzyme tests%Inhibitory concentration 50
目的 建立稳定的呼吸道合胞病毒(RSV)感染滴度测定的实时荧光定量PCR(Realtime Quantitative PCR,Q-PCR)检测方法和酶免疫斑点法(Enzyme Immunospots,EIS),并与半数定量方法(50% tissue culture infectious doses,TCID_(50))进行比较.方法 分别采用Q-PCR、EIS和TCID_(50)分析RSV感染的细胞和RSV攻毒后小鼠肺脏标本中的病毒滴度.结果 RSV感染细胞的上清中,三种分析方法检测的病毒滴度值之比为:Q-PCR和EIS(pfu)的比约为10:1,EIS(pfu)和TCID_(50)(TCID_(50) 换算成pfu)比约为10:1;RSV攻毒后小鼠肺脏标本中,三种分析方法检测的病毒滴度值之比为,QPCR和EIS(pfu)比约为1000:1,EIS(pfu)和TCID50(TCID50换算成pfu)比约为5:1.结论 成功建立了RSV感染滴度测定的Q-PCR和EIS分析方法,通过与TCID_(50)方法的比较分析,我们认为EIS法分析RSV滴度具有成本低和较高的灵敏度,有较好的应用前景.
目的 建立穩定的呼吸道閤胞病毒(RSV)感染滴度測定的實時熒光定量PCR(Realtime Quantitative PCR,Q-PCR)檢測方法和酶免疫斑點法(Enzyme Immunospots,EIS),併與半數定量方法(50% tissue culture infectious doses,TCID_(50))進行比較.方法 分彆採用Q-PCR、EIS和TCID_(50)分析RSV感染的細胞和RSV攻毒後小鼠肺髒標本中的病毒滴度.結果 RSV感染細胞的上清中,三種分析方法檢測的病毒滴度值之比為:Q-PCR和EIS(pfu)的比約為10:1,EIS(pfu)和TCID_(50)(TCID_(50) 換算成pfu)比約為10:1;RSV攻毒後小鼠肺髒標本中,三種分析方法檢測的病毒滴度值之比為,QPCR和EIS(pfu)比約為1000:1,EIS(pfu)和TCID50(TCID50換算成pfu)比約為5:1.結論 成功建立瞭RSV感染滴度測定的Q-PCR和EIS分析方法,通過與TCID_(50)方法的比較分析,我們認為EIS法分析RSV滴度具有成本低和較高的靈敏度,有較好的應用前景.
목적 건립은정적호흡도합포병독(RSV)감염적도측정적실시형광정량PCR(Realtime Quantitative PCR,Q-PCR)검측방법화매면역반점법(Enzyme Immunospots,EIS),병여반수정량방법(50% tissue culture infectious doses,TCID_(50))진행비교.방법 분별채용Q-PCR、EIS화TCID_(50)분석RSV감염적세포화RSV공독후소서폐장표본중적병독적도.결과 RSV감염세포적상청중,삼충분석방법검측적병독적도치지비위:Q-PCR화EIS(pfu)적비약위10:1,EIS(pfu)화TCID_(50)(TCID_(50) 환산성pfu)비약위10:1;RSV공독후소서폐장표본중,삼충분석방법검측적병독적도치지비위,QPCR화EIS(pfu)비약위1000:1,EIS(pfu)화TCID50(TCID50환산성pfu)비약위5:1.결론 성공건립료RSV감염적도측정적Q-PCR화EIS분석방법,통과여TCID_(50)방법적비교분석,아문인위EIS법분석RSV적도구유성본저화교고적령민도,유교호적응용전경.
Objective Respiratory syncytial virus (RSV) is the most common cause of lower respiratory infection in infants.It is very important to quantitative assay of RSV titer in the study on RSV pathogenesis,candidate vaccine and antiviral treatment.Therefore,we develop Real-time Quantitative PCR (Q-PCR) assay and enzyme immunospots (EIS) for titrating RSV and compare them with traditional 50% tissue culture infectious doses (TCID_(50)).Methods Q-PCR,based upon the RSV-L genes,and EIS were utilized to titrate samples from RSV culture supernatants and RSV infected mouse lungs.Then,the results were compared with TCID_(50).Results For the samples from RSV culture supernatants,the ratio of Q-PCR and EIS (plaque forming unit,pfu) was 10:1 and the ratio of EIS and TCID_(50) was 10:1 when TCID_(50) was converted as pfu.For the samples from RSV infected mouse lungs,the ratio of Q-PCR and EIS was 1000:1 and the ratio of EIS and TCID_(50) was 5:1.Conclusion We have successfully established Q-PCR and EIS to titrate RSV and compared them with TCID_(50).We concluded EIS is a cost-effective method to titrate RSV.
