山东省日照地区h9亚型aiv的分离 鉴定及ha基因的序列研究-isolation and identification of h9 subtype aiv and sequence study of ha gene in rizhao region of shandong province.docxVIP

山东省日照地区h9亚型aiv的分离 鉴定及ha基因的序列研究-isolation and identification of h9 subtype aiv and sequence study of ha gene in rizhao region of shandong province.docx

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山东省日照地区h9亚型aiv的分离 鉴定及ha基因的序列研究-isolation and identification of h9 subtype aiv and sequence study of ha gene in rizhao region of shandong province

determinationofhemagglutinationproperties,infectedchickenembryo,theaveragetimeofdeathofchickenembryoandtheICPI,thevirulencewasanalyzedandcompared.Theresultsshowthatthevirulencesofthereviruswereweakandthereweresmalldisparityinantigenicity.ThevirulenceofSDRZ2wasmuchstrongerthantheothers.TheHApotencyisbetween28and29.TheEID50isbetween10-6.15/0.2mLand10-8.69/0.2mL.TheEID50oftheSDRZ2is10-8.69/0.2mL.TherearesomedifferenceinMDTisbetween90.0hoursand103.2hours.AndtheSDRZ2poisonouschickenembryoaveragedeathtimemostisshortis90.0h,explainedthatthisviralthepathogenicityisstrong;Butothervirusschickenembryoaveragedeathtimeabove90h,showingthattheirpathogenicityisweak.Attacksthechicklingwhichthepoisonouschickenembryohatchesandattacksthepoisonouschicklingtoappearobviouslyhassplitexaminesthesymptom.PartII:CloningandandSequenceanalysisofcompleteHAgeneof4stainsofH9subtypeAIVThisresearchusedtheRT-PCRmethodtoincrease4H9hypotypeAIVHAgenefragment.Thesequentialanalysisresultindicatedthatamplificatedthegenefragmentlengthwhichis549bp.Thenucleotideandtheaminoacidsequencehomologyanalysisindicatedthattherelationofthese4stainsofH9subtypeAIVareclose,thehomologyabove93.7%,H9in2003separateswithShandongProvincethehypotypeAIVhomologyis94.5%.Differencehasnotthebigvariationalongwiththetimeandthearea.WithdomesticotherprovinceH9thehypotypeAIVhomologyis78.9%~96.9%.Thesystemcladogramanalysisresultindicatedthatthese4stainsofwithShandong,Guangzhou,Nanjing,theHebeiH9hypotypeAIVpoisonousbloodrelationshipisnear,belongstotheidenticalbranch.ItmightbeindicatedthatH9subtypeAIVoccurredinShandongarecloselyrelatedtothosepopularinGuangzhou,Nanjing,Hebei.Keywords:H9avianinfluenzavirus;BiologicalIdentification;HAgene符号说明英文英文全称中文缩写缩写AIAvianinfluenza禽流行性感冒AIVAvianinfluenzavirus禽流感病毒RNARibonucleicacid核糖核酸DNADeoxyribonucleicacid脱氧核糖核酸PCRPolymerasechainreaction聚合酶链式扩增反应RTReversetranscription反转录SPFSpecificpathogenfree无特定病原体TaqThermusaquaticu水生热栖酶AAAminoacids氨基酸LLiter升mLmilliliter毫升μLmicroliter微升bpbasepair碱基对dday天hhour小时minminute分钟

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