资源描述：
《冠状病毒Coronavirusl论文-2004 Real-Time Quantitative Fluorescent Reverse Transcriptase-PCR for Detection of Severe Acute Respiratory Syndrome-Ass.pdf》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。
1、OMolDiagn2004;8(4):231-235RIGINALRESEARCHARTICLE1084-8592/04/0004-0231/$31.00/0©2004AdisDataInformationBV.Allrightsreserved.Real-TimeQuantitativeFluorescentReverseTranscriptase-PCRforDetectionofSevereAcuteRespiratorySyndrome-AssociatedCoronavirusRNAWeijunChen,1,2Zuyua
2、nXu,1,3JingsongMu,4BoHe,2LingYang,1,2LinLin,5ShufangMeng,5FengMu,1,2HaixueGan,2ShengyongHuang,2JieWen,1,2JianqiuFang1,2andJianWang1,21BeijingGenomicsInstitute,ChineseAcademyofSciences,Beijing,China2BeijingBGI-GBIBiotechCo.Ltd,Beijing,China3InstituteofGeneticsandDevelo
3、pmentalBiology,ChineseAcademyofSciences,Beijing,China4Beijing302Hospital,Beijing,China5NationalInstitutefortheControlofPharmaceuticalandBiologicalProducts,Beijing,ChinaAbstractAim:SARS-associatedcoronavirus(SARS-CoV)hasbeenconfirmedasthepathogenforsevereacuterespirato
4、-rysyndrome(SARS).Theaimofourstudywastoconstructasensitiveandspecificreal-timequantitativefluorescent(QF)reversetranscriptase(RT)-PCRmethodforthedetectionofSARS-CoVRNA.Methods:Storedbloodspecimensfrom44patientswithconfirmedSARSwereusedalongwithbloodsamplesfromtwosetso
5、fcontrols,30healthyvolunteerswhohadnocontactwithSARSpatients,and30healthydoctorsandnurseswhohadcontactwithSARSpatientsbutwerewithoutsymptomsofSARS.TwopairsofprimersweresynthesizedbytheShanghaiSangonCompanyaccordingtoSARS-CoVBJ01strainsequence(AY278488),andthenapairofp
6、rimersweredesignedandcomparedwithapairofprimerspublishedbyWHO.Results:UsingserialdilutionsofSARS-CoV,the44bloodsamplesfromSARSpatientsspecimensweretested.Usinga0.01%dilutionofSARS-CoV,all44clinicalsamplestestedpositiveinourassay.Incomparison,usinga0.1%dilutionofSARS-C
7、oV,26ofthe44samplestestedpositiveusingtheWHOprimers.IntheQF-RT-PCRassay,therewasalinearamplificationfrom100copiesto108copiesofthecontrolRNAperRT-PCRandatleast10copies,andsometimeseven1copy,oftargetRNAtestedpositiveinourassay.Conclusion:Theprimerwedevelopedissufficient
8、lysensitiveandspecifictodiagnosesymptomaticSARS-CoVinfectionsandformonitoringvirusload.BythebeginningofNovember2003,theepide
