简介:Inordertoanalyzethenucleoprotein(NP)geneofCrimean-Congohemorrhagicfevervirus(CCHFV),viralRNAwasamplifiedbyRT-PCRbyusingtheproof-readingDNApolymerasetoproducethecompleteNPgene.ThePCRproductwassequenced,analyzedforphylogenesisandclonedintotheexpressionvectorpE132aandtherecombinantplasmidexpressedinE.coilBL-21withhighyield.Theprimarilypurifiedfusedprotein.wasusedtocoatELISAplatesforthedetectantibodies.ItwasfoundthesimilaritiesbetweenNPgeneofBA88166andotherXHFVsinnucleotidelevelandaminoacidcontentswereverysignificant,andtheNPgeneofBA88166encodedanucleoproteinwith482aminoacidandadeducedmolecularweight(MW)of54kDa.Westernblotassayshowedthatthefusionproteinexpressedinbacteriapossessedgoodantigenicity.TheresultswithELISAforthedetectionofthehumanandanimalseracollectedinendemicareaswerefoundtobeingoodaccordancetotheclinicaldiagnosis.ItconcludedthattherelationsofNPgenesofXHFVBA88166andotherXHFVsappearedtobeevolutionallyclose.Themethodologiesestablishedinthisstudywereaccurate,specific,rapidandreproduciblefortheclinicalexaminationsandepidemiologicalsurvey.
简介:TheZ10andZ37strainsofhemorrhagicfeverwithrenalsyndrome(HFRS)virusandtheMongoliangerbil(Merionsunguiculatus)kidneycellswereusedtopreparetheinactivatedbivalentvaccine.AphaseⅡclinicaltrialuseofthisvaccinewasmadein750Chinesevolunteers.Theresultsshowedthatthesidereactionratewas2.5%andthesero-conversionrateofneutralizingantibodiesagainstHantaanandSeoulvirusesintheinoculatedvolunteerswere87.6%and96.3%respectively.
简介:Inordertoelucidatethemolecularandimmunologicalmechanismsaswellasthepathogenesisofhemorrhagicfeverwithrenalsyndrome(HFRS),theCD8^+cytotoxicTlymphecytes(CTL)clonewasestablisheddirectlyfromperipheralbloodmononuclearcells(PBMC)ofpatientswithHFRS.TheactivitiesofCTLweredetectedasusualwithEBV-transformedlymphoblastoidcellline(BLCL)astargetcells.TheresultsshowedthattheCTLclonecouldrecognizedandkilledthetargetcellswithspecificityofnucleocapsidproteinofHantaanvirus(HTNVNP)withthecytotoxicitypercentagesof50.2%,25.4%and39.0%respectively.TheseresultsdemonstratedthattheantigenicepitopesofHTNVNPmainlylocatedontheC-temainaloftheviralnucleocapsidprotein.