簡(jiǎn)介：東北農(nóng)業(yè)大學(xué)碩士學(xué)位論文共表達(dá)GPMVF基因和GPVVP3基因重組禽痘病毒生物學(xué)特性研究姓名李丹申請(qǐng)學(xué)位級(jí)別碩士專業(yè)預(yù)防獸醫(yī)學(xué)指導(dǎo)教師王君偉20090620東北農(nóng)業(yè)大學(xué)農(nóng)學(xué)碩士學(xué)位論文IIIBIONOMICSSTUDIESOFRECOMBINANTFOWLPOXVIRUSESCOEXPRESSINGGPMVFGENEGPVVP3GENEABSTRACTINTHESTUDYTHEPRELIMINARYSTUDIESONTHEPHYSICALCHEMICALPROPERTIESOFRFPVVP3FGEICSTABILITYOFRFPVVP3FEXPRESSINGOFFGENEVP3GENEINCEFTHEEXPRESSINGOFFGENEVP3GENEINTHECHICKENWERECARRIEDOUTTHEVIRUSWASPURIFIEDTHENTHEEXISTENCEEXPRESSIONOFTHEFEIGNGENESWEREIDENTIFIEDBYPCRDOTELISAINTHESTUDYOFPHYSICALCHEMICALTHERECOMBINANTFOWLFOXVIRUSRGPVVP3FWHICHHASBEENTREATEDBYPHYSICALCHEMICALFACTSWEREINOCULATEDINTOCEFTHENTHEDIFFERENTINFECTIONTITERWEREOBSERVEDBETWEENTHETREATMENTUNTREATMENTVIRUSESPARTOFTHECHEMICALPROPERTYWEREVERIFIEDTHEOPTIMUMVIRALMULTIPLICATIONWEREDEFINITEDBYOBSERVEDTHEEFFECTSOFDIFFERENTCELLNUMBERDIFFERENTMOIWHENTHECELLDENSITYIS105CM2MOIIS001THETITEROFTHEVIRUSWILLACHIVEDTO308105100ULINTHEGEICSTABILITYSTUDIESOFRECOMBINANTFOWLPOXVIRUSTHERECOMBINANTFOWLPOXVIRUSONTHECEFWEREPASSAGEDF20GENERATIONSTHENTHE5TH10TH15TH20THGENERATIONSOFRECOMBINANTFOWLPOXWERECHOSENTOIDENTIFYTHEFGENEVP3GENE’SGEICSTABILITYTHERESULTSHOWNTHATBULEPLAQUEPERCENTAGEOFTHESEFOURGENERATIONIS100NOWHITEPLAQUEWEREOBSERVEDWHICHMAYOCCURWHENTHEFEIGNGENESWERELOSSUSETHE5TH10TH15TH20THGENERATIONSOFRECOMBINANTFOWLPOXVIRUSDNAASATEMPLATEFPCRAMPLIFICATION460BPFPARTLYAMPLIFIEDGENEFRAGMENT657BPVP3PARTLYGENEFRAGMENTWEREACQUIREDUSINGTHEOTHERTWOPAIRSOFPRIMERATOTALLENGTHOFTHEFGENETOTALLENGTHVP3GENEWEREAMPLIFIEDAFTERSEQUENCINGTHEGENEDEROF5TH10TH15TH20THGENERATIONSWERECOMPAREDWITHTHEGENEDEROFIGINALGENERATIONFGENEOFGPMVHADTWONUCLEOTIDECHANGEDBUTAMINOACIDDIDN’TCHANGEDVP3GENEOFGPVHADTHREENUCLEOTIDECHANGEDAMINOACIDDIDN’TCHANGEDTOOUSINGINDIRECTIMMUNOFLUESCENCETESTTOIDENTIFYTHEEXPRESSIONOFFGENEVP3GENEOFTHE5TH10TH15TH20THGENERATIONRECOMBINANTFOWLPOXVIRUSRESULTSFROMEXPERIMENTSHOWNTHATTHETWOFEIGNGENESOFTHERECOMBINANTFOWLPOXCANBESTABLYEXPRESSEDINCEFTHERECOMBINANTFOWLPOXHAVEGOODGEICSTABILITYTHEFEIGNGENEDIDN’TBELOSTINTHEPROCESSOFPASSAGEFGENEVP3GENECANEXPRESSCRESPONDINGPROTEINCRECTLY35DAYSOLDCHICKENWEREINOCULATEDWITHTHERFPVVP3FBY106PFUMLBLOODWERECOLLECTED7D14D21D28D35DAFTERIMMUNIZATIONTHESERUMWERESEPERATEDTHEANTIBODYLEVELSANTIGPVFWEREDETECTEDTHROUGHNEUTRALIZATIONTESTTHEANTIBODYLEVELS

簡(jiǎn)介：山東農(nóng)業(yè)大學(xué)碩士學(xué)位論文鴨副粘病毒分離鑒定、F基因序列分析及生物學(xué)特性研究姓名紀(jì)巍申請(qǐng)學(xué)位級(jí)別碩士專業(yè)預(yù)防獸醫(yī)學(xué)指導(dǎo)教師刁有祥20090614鴨副粘病毒分離鑒定、F基因序列分析及生物學(xué)特性研究膜出血、肺臟出血、十二指腸黏膜彌漫性出血。為確定發(fā)病原因，我們對(duì)死亡鴨胚、1日齡的病死鴨及出現(xiàn)神經(jīng)癥狀的雛鴨進(jìn)行了病原的分離、鑒定，確定為副粘病毒，命名為SDFCH株，對(duì)該分離株的生物學(xué)特性進(jìn)行了研究。毒力測(cè)定結(jié)果表明該病毒雞肝平均致死亡時(shí)間MDT為566H；1同齡SPF雞腦內(nèi)接種分離病毒致病指數(shù)ICPI為1786周齡SPF雞靜脈接種致病指數(shù)IVPI為259。攻毒雛鴨生長(zhǎng)緩慢，精神沉郁，剖檢可見(jiàn)氣管彌漫性出血、肺臟出血、腺胃乳頭出血、胰臟出血、十二指腸黏膜彌漫性出血，個(gè)別雛鴨腦膜出旭。第二部分鴨副粘病毒山東株F基因克隆與序列分析對(duì)20062008年從山東省不同地區(qū)規(guī)?；唸?chǎng)采集的疑似鴨副粘病毒病癥狀的病料進(jìn)行了鴨副粘病毒的分離鑒定及F基因克隆和序列分析，結(jié)果表明F基因的核苷酸序列較穩(wěn)定，不同地區(qū)6株鴨副粘病毒毒株的F基因全基因組序列均由1662BP組成，彼此間核苷酸序列同源性達(dá)953％一998％，親緣關(guān)系密切；與GENBANK已發(fā)表的鴨副粘病毒參考毒株的同源性介于874％978％；與GENBANK已發(fā)表的鵝副粘病毒參考毒株的同源性介于957～984％；與傳統(tǒng)的疫苗株LASOTA株的同源性在877％～883％之間；與F48E9的同源性在903‰910％之間。系統(tǒng)進(jìn)化樹(shù)分析表明這6株毒株與江蘇、黑龍江、廣東的鵝副粘病毒毒株和鴨副粘病毒毒株親緣性較近，屬于同一分支。本研究對(duì)所測(cè)定的6株鴨副粘病毒毒株F基因所對(duì)應(yīng)的氨基酸序列分析表明6株毒株之間的氨基酸同源性為953％～980％；與GENBANK已發(fā)表的鴨源副粘病毒的氨基酸同源性為8740／0975％；與鵝源副粘病毒的氨基酸同源性為964％～984％與傳統(tǒng)的疫苗株LASOTA株的氨基酸同源性為879％～883％；與F48E9的氨基酸同源性為901％91O％。6株毒株與鵝源副粘病毒的親緣性較近，因此推測(cè)鴨副粘病毒是由鵝源副粘病毒進(jìn)化而來(lái)。2

簡(jiǎn)介：東北農(nóng)業(yè)大學(xué)碩士學(xué)位論文鵝細(xì)小病毒98E鴨胚致弱株的培育及生物學(xué)特性研究姓名王春媛申請(qǐng)學(xué)位級(jí)別碩士專業(yè)獸醫(yī)學(xué)；預(yù)防獸醫(yī)學(xué)指導(dǎo)教師王君偉20120620ABSTRACTRESEARCHANDBIOLOGICALCHARACTERIZATIONOFADUCKEMBRYOATTENUATEDSTRAINOFGOOSEPARVOVIRUS98EABSTRACTGOOSEPARVOVLRUSINFECTIONALSOKNOWNASGOSLINGPLAGUEORDERZSY’SDISEASE，MAINLYINFECTS4DAYOLDTO20DAYOLDDOMESTICGOSLINGSANDMUSCOVYDUCKINGS．THESHORTCOURSEOFTHEDISEASE，RAPIDTRANSMISSION，ANDAHIGHMORTALITYRATE，CAUSINGSERIOUSECONOMICLOSSES，IT’SONEOFTHEIMPORTANTINFECTIOUSDISEASESOFGOOSECULTURALINDUSTRY．GGOSLINGPLAGUEWASPREVENTEDMAINLYTHROUGHIMMUNIZINGGOOSEWITHTHEGPVGOOSEEMBRYO／DUCKEMBRYOATTENUATEDVACCINEANDINACTIVATEDVACCINEANDTHENTHEGOSLINGACQUIREANTIBODYAGAINSTTHEGOSLINGPLAGUE．THISSTUDYUSEDTHECULTIVATEGOOSEPARVOVIRUSDUCKEMBRYOATTENUATEDSTRAINGPV98D15WITHCONTINUOUSLYPASSAGINGIN9DAYOLDSPFDUCKEMBRYO，IDENTIFIEDTHEVIRALNUCLEICACIDOFTHEDIFFERENTGENERATIONSBYPCRANDDETERMINEPARTSOFBIOLOGICALCHARACTERISTICSDURINGTHEGENERATIONPROCESS．THERESULTSOFTHEBIOLOGICALCHARACTERISTICSOFGOSLINGPLAGUEDUCKEMBRYOATTENUATEDSTRAINSGPV98D15SHOWEDTHATDUCKEMBRYOCOULDNOTDIEFROMFITOF10，STARTGRADUALLYDIEDFROMF11TOF13．BEGINNINGWITHF14，LETHALITYRATEOFDUCKEMBRYOKEEPSTABLEAT100％．THELETHALTIMEOFDUCKEMBRYOREPRESENTSHORTREGULARLY．THEDEADBODIESOFDUCKEMBRYOWERECONGESTED，ANDTHELOCUSOFHEMORRHAGEMAINLYCONCENTRATEDINTHEHEAD，NECK，CHESTANDLEGS，WHICHARECONSISTENTW．ITHTHEFEATUREOFGPVPATHOLOGICALCHANGES．ITWASSUCCESSFULLYCLONEDTHEGPV98D15STRAINNSLGENEANDVPLGENEBYPCR；THEVPLGENEOFGPV98D15STRAINANDGPV98ESTRAINHADPARTIALNUCLEOTIDESEQUENCEMUTATION．THEGPV98D15STRAINATTENUATEDSTABILITYANDSAFETYBYDUCKEMBRYOLETHALDOSE，THEPATHOGENICITYTESTOFGOOSEEMBRYOANDSAFETYTEST．GPV98D15VIRUSCOULDINFECTDUCKEMBRYOFIBROBLASTS．ITCANBEUSEDDUCKEMBRYOFIBROBLASTSINFECTEDANDHALFTHEAMOUNTOFINDIRECTIMMUNOFLUORESCENCEIFATODETERMINATION．WECOMPAREDTHEIMMUNOGENICISONOFARENUATEDSTRAINSWITHTHECOMMERCIALVACCINESSYG4150．IMMUNING20DAYOLDGOSLINGSWITHGPV98DJ5STRAINORSYG4150，ANTIBODYLEVELRISESCEASELESSLY，REACHEDAPEAKATFIFTHWEEKS．ANDTHENDECLINESLOWLYBYELISA，AGARDIFFUSIONTESTANDNEUTRALIZATIONTEST．ANTIBODYLEVELOFGPV98D15IMMUNEGROUPISSLIGHTLYLOWCOMPAREDWITHCOMMERCIALVACCINESYG41～50，BUTTHEREISNOSIGNIFICANTDIFFERENCEFPD．05．DETECTIONOFIMMUNEGPV．98D15STRAINTOMAKESMALLGOOSEPRODUCESGPVSPECIFCNEUTRALIZINGANTIBODYBYNEUTRALIZATIONTEST．THEGOSLINGSACQUIREDPROTECTIVEIMMUNITY．THEREFORE，THERESEARCHPROVIDEDASAFE，EFFECTIVE，NOSEASONLIMITATIONOFCANDIDATESTRAINS，ITALWAYSTHEMATERIALFOUNDATIONFORGOSLINGPLAGUEVIRUSAFIENUATIONMECHANISMRESEARCH．．KEYWORDSGOSLINGPLAGUE；ATTENUATEDSTRAIN；SAFETY；IMMUNOGENICITYII

簡(jiǎn)介：中山大學(xué)博士學(xué)位論文石斑魚虹彩病毒（SGIV）囊膜蛋白組學(xué)及兩個(gè)新結(jié)構(gòu)蛋白基因的鑒定和功能初步研究姓名萬(wàn)晴姣申請(qǐng)學(xué)位級(jí)別博士專業(yè)生物化學(xué)與分子生物學(xué)指導(dǎo)教師秦啟偉20090606萬(wàn)晴姣石斑魚虹彩病毒SGIV囊膜蛋自組學(xué)及兩個(gè)新結(jié)構(gòu)蛋自基兇的鑒定和功能初步研究VP90在病毒感染過(guò)程中可能起重要作用。為了弄清病毒結(jié)構(gòu)蛋白在病毒感染中的作用，結(jié)合生物信息學(xué)分析，我們選取ORF038L作為候選基因進(jìn)行研究，SGIVORF038L編碼170個(gè)氨基酸，含有一個(gè)保守的RGD結(jié)構(gòu)域。從SGIV全基兇組中克隆了ORF038全長(zhǎng)基因，進(jìn)行了原核表達(dá)，并免疫小鼠制備出抗體。轉(zhuǎn)錄表達(dá)時(shí)序分析表明，SGIVORF038L在感染細(xì)胞后8H開(kāi)始轉(zhuǎn)錄，24H開(kāi)始表達(dá)。結(jié)合藥物抑制實(shí)驗(yàn)，揭示ORF038是一個(gè)晚期基因。免疫熒光顯微鏡觀察，結(jié)果顯示VP38在感染細(xì)胞中主要聚集在病毒加工廠周圍。WESTERNBLOT和免疫電鏡結(jié)果表明VP38定位于病毒衣殼，是一種病毒衣殼蛋白。抗體中和試驗(yàn)和PUL1DOWN實(shí)驗(yàn)結(jié)果表明SGIVVP38在病毒感染過(guò)程中可能起重要作用。關(guān)鍵詞SGIV；蛋白質(zhì)組學(xué)；囊膜蛋白；衣殼蛋白；亞細(xì)胞定位；抗體中和N

簡(jiǎn)介：山東農(nóng)業(yè)大學(xué)博士學(xué)位論文鴨圓環(huán)病毒的分子流行病學(xué)調(diào)查及診斷方法的建立姓名張興曉申請(qǐng)學(xué)位級(jí)別博士專業(yè)預(yù)防獸醫(yī)學(xué)指導(dǎo)教師柴同杰姜世金20090616鴨圓環(huán)病毒的分子流行病學(xué)調(diào)查及診斷方法的建立3、櫻桃谷鴨人工感染鴨圓環(huán)病毒的病理動(dòng)態(tài)變化本研究以已經(jīng)完成全基因組序列測(cè)定的LY0701株DUCV為研究材料，利用感染LY0701株的陽(yáng)性病料人工接種DUCV陰性的3周齡健康雛鴨，分別于攻毒后第2、4、6、8周對(duì)發(fā)病鴨進(jìn)行病理觀察。結(jié)果表明，與對(duì)照組相比，LY0701株陽(yáng)性病料感染的雛鴨臨床癥狀均出現(xiàn)發(fā)育不良，消瘦，羽毛凌亂等現(xiàn)象。剖檢顯示胸腺、法氏囊、脾臟、肝臟不同程度出血、腫脹，其它內(nèi)臟器官病變不明顯。病理切片顯示胸腺、法氏囊、脾臟、肝臟出血，淋巴細(xì)胞壞死、崩解等變化。4、DUCVCAP蛋白基因原核表達(dá)及產(chǎn)物免疫原性研究根據(jù)已發(fā)表的DUCVLY0701株CAP基因序列，設(shè)計(jì)一對(duì)引物，以測(cè)序正確的LY0701株CAP基因質(zhì)粒為模板，擴(kuò)增出相應(yīng)的基因片段，以PGEX6P1為載體進(jìn)行連接，轉(zhuǎn)化大腸桿菌BL21株，挑選陽(yáng)性克隆，測(cè)序驗(yàn)證正確后，分別進(jìn)行原核表達(dá)。SDSPAGE電泳表明，表達(dá)產(chǎn)物的條帶大小與理論值相符。利用原核表達(dá)產(chǎn)物免疫BALB／C小鼠，制備抗CAP蛋白的血清，標(biāo)記為ABCAP。以FITC標(biāo)記的羊抗鼠IGG為二抗，建立了間接免疫熒光染色法IFA。利用ABCAP對(duì)人工感染病料的檢測(cè)表明，在脾臟、法氏囊、胸腺、肝臟的冷凍切片中可檢測(cè)到被熒光染色的完整的細(xì)胞；在心臟、肺臟、腎臟等病料中未檢測(cè)到熒光染色的陽(yáng)性細(xì)胞。為進(jìn)一步確定IFA檢測(cè)的敏感性，臨床上分別收集疑似DUCV感染的96只鴨子的胸腺、法氏囊、脾臟、肝臟、骨髓，利用IFA進(jìn)行DUCV檢測(cè)。結(jié)果表明，當(dāng)鴨子感染DUCV后，脾臟的病毒檢出率最高，達(dá)到1875％，其它臟器的病毒檢出率從高到低依次為法氏囊、肝臟、胸腺。利用ABCAP對(duì)臨床病例檢測(cè)表明，該方法更適合用于8周齡以上鴨子的脾臟檢測(cè)。5、DUCV在不同組織及細(xì)胞中培養(yǎng)的探討對(duì)PCR檢測(cè)DUCV強(qiáng)陽(yáng)性的5份病料LQ76，LQ66，LQL4，LY5，WS23進(jìn)行處理并1通過(guò)鴨胚的卵黃囊、絨毛尿囊膜、尿囊腔等組織進(jìn)行接種，2鴨胚成纖維細(xì)胞、鴨胚肝細(xì)胞、鴨胚腎細(xì)胞、鴨胚脾細(xì)胞接種陽(yáng)病料組織，將收獲的組織和細(xì)胞2

簡(jiǎn)介：揚(yáng)州大學(xué)博士學(xué)位論文感染馬立克氏病病毒SPF雞法氏囊蛋白質(zhì)組學(xué)研究姓名盧占軍申請(qǐng)學(xué)位級(jí)別博士專業(yè)基礎(chǔ)獸醫(yī)學(xué)指導(dǎo)教師秦愛(ài)建20090501II揚(yáng)州大學(xué)博士學(xué)位論文重萎縮，濾泡內(nèi)淋巴細(xì)胞排列松散，濾泡間質(zhì)增寬，肌細(xì)胞層彌漫性增生。同時(shí)利用半定量RTPCR技術(shù)檢測(cè)不同階段法氏囊中MDV的GB和MEQ基因的MRNA相對(duì)表達(dá)水平。結(jié)果顯示MDVMEQ基因的MRNA水平在7DPI、21DPI時(shí)明顯增高，21DPI時(shí)達(dá)到差異極顯著；MDVGB基因在4DPI和21DPI時(shí)顯著性增高，而在潛伏期7DPI和14DPI時(shí)則明顯下降，這些數(shù)據(jù)顯示MDV在感染雞法氏囊中復(fù)制正常，也為后期的病毒感染宿主差異蛋白質(zhì)組學(xué)研究提供了條件。3馬立克氏病病毒感染后法氏囊組織的蛋白質(zhì)組學(xué)研究采用2DE技術(shù)，對(duì)RBLB感染的SPF雞和對(duì)照組雞的法氏囊在不同感染階段4DPI、7DPI、14DPI和2LDPI的組織樣品進(jìn)行了蛋白質(zhì)組分離，利用PDQUEST801軟件對(duì)2一DE圖譜進(jìn)行分析，對(duì)其中的部分差異點(diǎn)進(jìn)行質(zhì)譜鑒定。結(jié)果平均每塊膠能檢測(cè)到800900個(gè)蛋白點(diǎn)，四個(gè)階段共有77個(gè)差異蛋白點(diǎn)，利用統(tǒng)計(jì)分析方法篩選出差異極顯著且變化有規(guī)律的蛋白點(diǎn)29個(gè)，其中表現(xiàn)為持續(xù)上調(diào)的有13個(gè)，持續(xù)下調(diào)的有10個(gè)，先期上調(diào)后期下調(diào)的2個(gè)，先期下調(diào)后期上調(diào)的4個(gè)。對(duì)這些蛋白點(diǎn)進(jìn)行切膠消化，用基質(zhì)輔助激光解吸電離飛行時(shí)間質(zhì)譜MATRIXASSOCIATEDLASERDISSOCIATION／IONIZATIONTIMEOFNIGLL仃NASSSPEC仃OMETMMALDITOFMS進(jìn)行鑒定分析。共鑒定出24個(gè)蛋白，它們是載脂蛋白AIAPOAI、白血病相關(guān)蛋白18LAPL8、癌蛋白18OPL8、脅相關(guān)蛋白R(shí)ABLLA、兩種熱休克蛋白27HSP27、BUB同系蛋白、含硫氧還蛋白域5TNDC5、伴侶素蛋白CCT、D2微球蛋白BMG、熱休克蛋白40HSP40、磷酸甘露糖變位酶蛋白、慢骨骼肌肌鈣蛋白T、蛋白質(zhì)二硫化物異構(gòu)酶A3前體、DUTP焦磷酸酶蛋白、D樣不均一核核糖核蛋白、醛縮酶C蛋白、胰蛋白、LOCL29607類似蛋白、含錯(cuò)配修復(fù)酶假定蛋白、含GTP嬲E假定蛋白、兩種含GAL‰E1樣結(jié)構(gòu)域假定蛋白。另外，還發(fā)現(xiàn)了兩個(gè)數(shù)據(jù)庫(kù)中沒(méi)有的禽源蛋白，這有待于更進(jìn)一步的研究。通過(guò)數(shù)據(jù)庫(kù)查詢和生物信息學(xué)分析，證實(shí)這些已知的差異蛋白與新陳代謝相關(guān)的占19％，與蛋白質(zhì)折疊相關(guān)的占15％，與細(xì)胞增殖和調(diào)亡相關(guān)的占15％，與信號(hào)轉(zhuǎn)導(dǎo)相關(guān)的占22％，與免疫相關(guān)的占15％，與細(xì)胞骨架結(jié)構(gòu)相關(guān)蛋白的占7％，其它占7％。在此基礎(chǔ)上結(jié)合現(xiàn)有的文獻(xiàn)報(bào)道，對(duì)這些差異蛋白質(zhì)的功能、與病毒感染發(fā)病關(guān)系以及腫瘤相關(guān)方面進(jìn)行分析討論。提出APOAI持續(xù)的過(guò)量表達(dá)可能與MDV病毒粒子跨膜感染機(jī)制有關(guān)，同時(shí)以APOAI過(guò)表達(dá)為主導(dǎo)的脂類代謝紊亂可能是最終導(dǎo)致MDV感染易引起動(dòng)脈粥樣硬化的主要原因；L廿18和OPL8的波動(dòng)性調(diào)節(jié)可能與MDV引起的腫瘤形成早期有關(guān)；RBLB誘導(dǎo)的HSP27蛋白過(guò)量表達(dá)可能有利于其子代病毒大量產(chǎn)生持續(xù)上調(diào)表達(dá)的R盈B11A蛋白可能與MDV細(xì)胞內(nèi)轉(zhuǎn)運(yùn)，囊泡出芽，逃避免疫或復(fù)制機(jī)制有關(guān)，RABLLA也有

簡(jiǎn)介：河南農(nóng)業(yè)大學(xué)碩士學(xué)位論文河南省豬乙型腦炎血清流行病學(xué)調(diào)查及乙型腦炎病毒河南地方株的分離鑒定姓名崔光輝申請(qǐng)學(xué)位級(jí)別碩士專業(yè)預(yù)防獸醫(yī)學(xué)指導(dǎo)教師陳陸20090501河南農(nóng)業(yè)大學(xué)2009屆獸醫(yī)碩士畢業(yè)論文IDENTIFICATIONANDISOLATIONOFJAPANESEENCEPHALITISVIRUSANDTHEPIGBLOODSERUMEPIDEMIOLOGYINVESTIGATIONOFHENANPROVINCESUPERVISORPROFCHENLUMASTERCANDIDATECUIOUAN曲UIABSTRACTJAPANESEENCEPHALITISWASONEKINDOFCENTRALNERVOUSSYSTEM’SACUTEINFECTIOUSDISEASEAFTERMOSQUITOBITEDISSEMINATIONTHREATENSERIOUSLYHUMANANDANIMALSHEALTH111EPIGWASJAPANESEENCEPHALITISVIRUS’SIMPORTANTSTORAGEHOSTANDINCREASESTHEHOST，ANDWASIT’SMAINORIGINOFINFECTIONTOPREVENTJAPANESEENCEPHALITIS’SUNPOPI_TLARITYEFFECTIVELYPROTECTSTHEHUMANANDANIMALSSECURITYHASCONDUCTEDTHEINVESTIGATIONANDSTUDYTOTHEHENANPROVINCEPIGINFECTIONENCEPHALITISVIRUSSITUATIONTHISEXPERIMENTMAKEDTHEAPPRAISALFIRSTFROMTHESEROLOGYASPECTTOJAPANESEENCEPHALITISVIRUS’SIMMUNITYPROTECTIONSITUATION，ALSOHADCARRIEDONTHISVIRUS’SSEPARATIONAPPRAISALFROMTHEMOLECPLARBIOLOGYASPECTTOTHESUSPICIOUSCASESINCEMAY’2008，MYLABORATORYHADCOLLECTED801BLOODSERUMSFROM12CITYCERTAINPIGFARMSWHICHALREADYIMMUNITIEDJAPANESEENCEPHALITISOFHENANPROVINCE，THEJEVIGGIMMUNEBODYWEREEXAMINATIEDBYPIGJAPANESEENCEPHALITISVIRUSELISAIMMUNEBODYEXAMINATIONREAGENTBOXANDCARRIEDONTHEANALYSISPIGJAPANESEENCEPHALITISPOPTLARSITUATIONSOFHENANPROVINCEDURING2008YEARTHE22DOUBTFLALJAPANESEENCEPHALITIS’SMATERIALWERESEPERATEDANDCHARACTRIZEDFROMSICKNESSMORBIDITYPIGFARM’SOFTHEHENANPROVINCEVARIOUSAREAWITHCELLCLALTURE’SMETHODSEPARATIONVIRUS，CARRIESONTHERTPCRAPPRAISALTOTHENEWSEPARATION’SVIRUSTHERESULTISASFOLLOWS1GTHEJAPANESEENCEPHALITISANTISERUMBODYMASCI_TLINERATEWAS4794％OFOUTPROVINCE，INSOMEAREASPIGFARMSTHEIMMUNEBODYMASCLALINERATEWASONLY1O00％BECAUSEAFTEREXAMINESTHEBLOODSERUMISTHEVACCINEIMMUNITYBLOODSERUM，THEREFORE，ENTIREPROVINCEPIGFARMIMMUNITYPROTECTIONRATIOINSUFFICIENT50％，ISGENERALLYLOWWHICHALEINHIGHRISKSENDTHEAREAT至DIFFERENTSEASONSFROMTHECONTRAST，IN56MONTHTHEPOSITIVERATEOFJAPANESEENCEPHALITISWASLOWLESSTHAN40％，ANDINAUGUSTTHEHIIGHESTSERUMANTIBODYPOSITIVERATE，REACHING5599％，F(xiàn)OLLOWEDINJPLYREACHING4750％，WHICHAREPROBABLYFARMSINGENERALINTHE

簡(jiǎn)介：福建農(nóng)林大學(xué)碩士學(xué)位論文雙生病毒的檢測(cè)及對(duì)煙粉虱生物學(xué)特性的影響姓名歐陽(yáng)智剛申請(qǐng)學(xué)位級(jí)別碩士專業(yè)植物病理學(xué)指導(dǎo)教師王聯(lián)德20100401福建農(nóng)林大學(xué)碩士學(xué)位論文STUDYOILDETECTIONOFGEMINIVIRUSANDITSIMPACTONTHEBIOLOGICALCHARACTERISTICSOFBEMISIATABACIABSTRACTGEMINIVIRUSESTESTEDINTHISPAPERINVOLVEDRAMIEMOSAICVIRUSRAMOV，PAPAYATEAFCURLGUANGDONGVIRUSPALCUGUVANDAGERATUMYELLOWVEINVIRUSAYVV．WESTUDIEDTHETRANSMISSIONOFTHOSETHREEGEMINIVIRUSBYBEMISIATABACIBBIOTYPE．WESTUDIEDTHEIMPACTSOFPALCUGUVONTHEDEVELOPMENTTIME，F(xiàn)ECUNDITY,LONGEVITYANDFEEDINGAMOUNTOFB．TABACIUNDERTHEDIFFERENTTEMPERATURESANDHOSTPLANTS．THEMAJORFINDINGSAREASFOLLOWS1B．TABACICANTRANSMITRAMOV,PALCUGUV,AYVVANDAVVVPEFFICIENTLY,THOSEGEMINIVIRUSESCOULDINJECTEDDIFFERENTSPECIESOFHEALTHYTOBACCOAFTERTRANSMISSION，ANDTHENTOBACCOSHOWEDTHESYMPTOMOFGEMINIVIRUSES．2THEDEVELOPMENTTIMESOFINFECTEDB．TABACIWERELONGERTHANTHATOFHEALTHYB．TABACIUNDERTHETEMPERATURESTESTED．THEDIFFERENCESWERESIGNIFICANTUNDERTHEFORMERFOURTEMPERATURES．THEOPTIMUMTEMPERATURESOFINFECTEDANDHEALTHYB．TABACIWEREBOTH25TO28℃，ANDDEVELOPMENTTIMESWERESHORTERTHANTHATOFOTHERTEMPERATURES．3THEDEVELOPMENTTIMESOFDIFFERENTSTAGESOFINFECTEDB．TABACIONAGERATUMCONYZOIDESWERELONGERTHANTHATOFHEALTHYB．TABACIONIPOMOEABATATASLAM，RESPECTIVELYEXCEPTED2加INSTARNYMPHS，ANDTHERESULTOFHEALTHYB．TABACIWASSAME．4THEHATCHINGRATEOFINFECTEDB．TABACIWEREHIGHERTHANTHATOFHEALTHYB．TABACI，HOWEVERTHEREWASNOSIGNIFICANTDIFFERENCEEXCEPTEDUNDERTHECONDITIONOF31℃．THELIFETIMEOFINFECTEDFEMALEB．TABACIWASLONGERTHANTHATOFHEALTHYFEMALEB．TABACI,ANDTHEFECUNDITYOFINFECTEDFEMALEB．TABACIWASALSOMORETHANTHATOFHEALTHYFEMALEB．TABACIAT26。C．THEAREAOFHONEYDEWOFINFECTEDB．TABACIWEREALLSMALLERTHANTHATOFHEALTHYB．TABACIUNDERDIFFERENTTEMPERATURECONDITIONSONAGERATUMCONYZOIDES．THEE位CTSOFVIRUSANDTEMPERATUREONFEEDINGAMOUNTOF2

簡(jiǎn)介：山東農(nóng)業(yè)大學(xué)碩士學(xué)位論文兩個(gè)馬鈴薯Y病毒分離物的生物學(xué)和分子特性姓名王彪申請(qǐng)學(xué)位級(jí)別碩士專業(yè)植物病理學(xué)指導(dǎo)教師李向東20090608兩個(gè)馬鈴薯Y病毒分離物的生物學(xué)和分子特性ABSTRACTPOTATOVIRUSYIFVⅥISONEOFTHEMOSTWIDESPREADVIRUSESANDCAUSESHUGELOSSESTOSOLANACEOUSCROPSINCLUDINGPOTATO，TOBACCO，EGGPLANTANDTOMAT0TWOPVYISOLATES，AQ4ANDFZ10，WEREISOLATEDFROMTOBACCOPLANTSINAIQIUANDTAI’ANOFSHANDONGPROVINCEBOTHISOLATESCOULDINDUCEMOSAICANDMOTTLEIN12TOBACCOCULTIVARSINCLUDINGZHONGYAN100THECOMPLETEGENOMICSEQUENCESOFAQ4ANDFZIOWEREDETERMINEDFROMSEVENOREIGHTOVERLAPPINGCDNACLONESOBTAINEDFROMRTPCRAND5RACEEXCLUDINGTHEPOLYATAIL，THEGENOMEOFAQ4ANDFZL0WERE9700AND9699NUCLEOTIDESINLENGTH，RESPECTIVELYBOTHOFTHEMENCODEDAPOLYPROETINOF3061AMINOACIDS，WHICHWASTHENCLEAVEDINTOTENPRODUCTSBYTHREEVIRUSENCODEDPROTEASESAQ4SHAREDHIGHESTIDENTITYOF973％WITHISOLATE2614ATNUCLEOTIDELEVEL，WHILEFZL0SHAREDHIGHESTIDENTITYOF952％WITHPVY12BOTHAQ4ANDFZL0WERERECOMBINANTSOFNANDOPARENTSTHENUCLEOTIDESNT1496AND24979700OFAQ4GENOMEWEREFROMISOLATEOZPVF’，WHILENT4972496FROMISOLATECH605PVYSTHENT1496AND24976533OFFZ10GENOMEWEREFROMISOLATESASA110PVN，WHILENT4972496AND65349699WERE￡舊MISOLATECH605PVYN碭EPHYLOGENETICANALYSISRESULTSSHOWEDTHATAQ4WASCLUSTEREDTOSTRAINPVYN0WHILEFZL0PVYN州BOTHOFTHEMHADN邯EHCPROHOWEVER，NEITHEROFTHEMINDUCEDNECROSISONTOBACCOTHESEREULSTSIMPLIEDTHATTHEREEXISTEDOTHERMOTIFBESIDESHCPREGULATINGTHEVEINALNECROSISSYMPTOMINPVYGENOMEKEYWORDSPOTATOVIRUSY；GENOME；MOTIFIREEOMBINANTION；HCPRO2

簡(jiǎn)介：東北農(nóng)業(yè)大學(xué)碩士學(xué)位論文表達(dá)牛輪狀病毒VP7蛋白重組干酪乳桿菌的構(gòu)建及其免疫學(xué)初步分析姓名張冠群申請(qǐng)學(xué)位級(jí)別碩士專業(yè)預(yù)防獸醫(yī)學(xué)指導(dǎo)教師李一經(jīng)20090620東北農(nóng)業(yè)大學(xué)農(nóng)學(xué)碩士學(xué)位論文VICONSTRUCTIONOFRECOMBINANTLACTOBACILLUSCASEIEXPRESSIONVP7OFBOVINEROTAVIRUSIMMUNOGENICITYANALYSISABSTRACTBOVINEROTAVIRUSBELONGEDTOREOVIRIDAEFAMILYROTAVIRUSGENUSITWASTHEMAJPATHOGENOFCALFNONBACTERIALDIARRHEATHEVIRUSUSUALLYINFECT1540DAYAGECALVESTHECLINICALSYMPTOMAREDEPRESSEDEMESIALOSEAPPETITEDIARRHEASEVEREDEHYDRATIONACIDPOISONINGIFITWASACCOMPANYINFECTEDBYBOVINECONAVIRUSBACTERIATHEMTALRATECANREACHUPTO80THISDISEASEHASCOSTCATTLEPRODUCERSSERIOUSLOSSTHISDISEASEBELONGEDTOINTESTINALINFECTIONITSPREADTHROUGHFECALALROUTESODESIGNINGAVACCINEASTHEFIRSTLINEOFDEFENSEETIOLOGICALFACTINVADINGGANISMBYSTIMULATINGMUCOSAIMMUNITYHASIMPTANTSIGNIFICANCEINTHISSTUDYLACTOBACILLUSCASEI393WASEDASANANTIGENDELIVERYVEHICLETHEMAINPROTECTIVEANTIGENVP7WASEDASTHETARGETGENETHEIVEGENEVP7WASDIRECTIONALEDINTOTHESURFACEEXPRESSIONVECTPPG1SECRETARYEXPRESSIONVECTPPG2CONSTRUCTEDRECOMBINANTSTRAINSPPG1VP7LCASEI393PPG2VP7LCASEI393RECOMBINANTLACTOBACILLUSCASEICANEXPRESSTRANSMITEXOGENOUSANTIGENITSELFCANATTACHTOTHEININTESTINALTRACTBEARBILESALTPANCREATICENZYMESOITCANSTIMULATEMUCOSAIMMUNOLOGICSYSTEMOFINTESTINALTRACTTHERECOMBINANTSTRAINSCONSTRUCTEDINTHISSTUDYUSEDASALVACCINECANPREVENTCALFINFECTINGBOVINEROTAVIRUSTHISSTUDYAMPLIFICATEDVP7GENEOFBOVINEROTAVIRUSINTHEDIARRHEASTOOLOFCALFTHERESULTOFSEQUENCINGWASTHATTHEGENEBELONGEDTOG6SEROTYPESPREADWIDELYATFIRSTWEUSEECOLIEXPRESSIONSYSTEMTOEXPLETHEEXPRESSIONCONDITIONINPROKARYOTICEXPRESSIONSYSTEMTHEIMMUNITYEFFECTIVETHERESULTISTHAT51254POSITIONSOFVP7PROTEINWHICHINCLUDEDMAJANTIGENSCANEXPRESSWELLMEOVERTHEANTISERUMOFMOUSEHADHIGHERELISATITER（16400）NEUTRALIZINGANTIBODYTITER（1316）INTHISFOUNDATIONWECONSTRUCTEDSURFACEEXPRESSIONSYSTEMPPG1VP7LCASEI393SECRETARYEXPRESSIONSYSTEMPPG2VP7LCASEI393THATCANEXPRESSVP7PROTEINOFBOVINEROTAVIRUSTHERECOMBINANTSTRAINSCONSTRUCTEDINTHISSTUDYWEREINDUCEDBYLACTOSETHEBACTERIALYSATECULTIVATESUPERNATANTWASDETECTEDBYWESTERNBLOTSHOWEDTHATPURPOSEPROTEINWASDETECTEDONTHEBACTERIALYSATEOFPPG1VP7LCASEI393PPG2VP7LCASEI393WEALSODETECTEDTHEPROTEINONTHECULTIVATESUPERNATANTOFPPG2VP7LCASEI393THEINDIRECTIMMUNOFLUESCENCETESTSHOWEDTHATTHEEXPRESSEDPROTEINWASONTHESURFACEOFPPG1VP7LCASEI393TOIDENTIFYWHETHERTHERECOMBINANTSTRAINSHAVETHEABILITYTOINDUCESYSTEMICMUCOSAL

簡(jiǎn)介：MOLECULAREPIDEMIOLOGYSURVEYOFCANINEDISTEMPER，CANINEPARVOVIRUSINFECTIONANDCANINECORONAVIRUSINBEIJINGBYZHANGJINSUPERVISORSPROFHOUJIAFAASSOCIATERESEARCHERYANGBINGATHESISSUBMITTEDTONANJINGAGRICULTURALUNIVERSITYINPARTIALFULFILLMENTOFTHEREQUIREMENTSFORTHEDEGREEOFMASTEROFTHEAGRONOMYCOMPLETEDINAPR，2009COMMENCEMENTINJUN，2009原創(chuàng)性聲明JJLLLLL11LLLL1LLLIIJ＼1763105本人鄭重聲明所呈交的學(xué)位論文，是本人在導(dǎo)師的指導(dǎo)下，獨(dú)立進(jìn)行研究工作所取得的成果。除文中已經(jīng)注明引用的內(nèi)容外，本論文不包含任何其他個(gè)人或集體己經(jīng)發(fā)表或撰寫過(guò)的作品成果。對(duì)本文的研究做出重要貢獻(xiàn)的個(gè)人和集體，均已在文中以明確方式標(biāo)明。本人完全意識(shí)到本聲明的法律結(jié)果由本人承擔(dān)。學(xué)位論文作者需親筆簽名編葉每移6具帕學(xué)位論文版權(quán)使用授權(quán)書本學(xué)位論文作者完全了解學(xué)校有關(guān)保留、使用學(xué)位論文的規(guī)定，同意學(xué)校保留并向國(guó)家有關(guān)部門或機(jī)構(gòu)送交論文的復(fù)印件和電子版，允許論文被查閱和借閱。本人授權(quán)南京農(nóng)業(yè)大學(xué)可以將本學(xué)位論文的全部或部分內(nèi)容編入有關(guān)數(shù)據(jù)庫(kù)進(jìn)行檢索，可以采用影印、縮印或掃描等復(fù)制手段保存和匯編學(xué)位論文。保密口，在年解密后適用本授權(quán)書。本學(xué)位論文屬于不保茸西請(qǐng)?jiān)谝陨戏娇騼?nèi)打“√”學(xué)位論文作者需親筆簽名勿錈導(dǎo)師需親筆簽刎年毛只|7黽緲呷年彩月，孑日

簡(jiǎn)介：西南大學(xué)碩士學(xué)位論文豬瘟病毒E2基因RTPCR檢測(cè)方法的建立及病毒分子流行病學(xué)研究姓名湯波申請(qǐng)學(xué)位級(jí)別碩士專業(yè)預(yù)防獸醫(yī)學(xué)指導(dǎo)教師黃偉王琴20090601組織器官進(jìn)行免疫組化檢測(cè)抗原；同時(shí)進(jìn)行臨床觀察、體溫測(cè)定并給予進(jìn)行臨床計(jì)分。RTPCR檢測(cè)結(jié)果顯示，感染后第一天有7種組織器官CSFV檢測(cè)為陽(yáng)性，至第五天34種急性樣品全部檢測(cè)為陽(yáng)性，整個(gè)感染病程中血液的CSFV核酸檢出數(shù)最多，檢出率93．70／015／16其次是各種與免疫相關(guān)的淋巴結(jié)和淋巴樣組織，而肌肉和皮膚檢出數(shù)最少；4種體外樣品糞、尿、眼分泌物、鼻拭子分別在3．5天內(nèi)檢出陽(yáng)性。免疫組化檢測(cè)結(jié)果顯示，22種組織器官切片中有19種觀察到陽(yáng)性信號(hào)，這19個(gè)組織器官與CSFVRTPCR檢測(cè)結(jié)果相符合；而皮膚、肌肉和喉頭的組織切片中未觀察到陽(yáng)性信號(hào)。結(jié)果表明在CSFV人工急性感染模型中，病毒感染后能在24小時(shí)間內(nèi)出現(xiàn)病毒血癥，持續(xù)整個(gè)急性感染病程；病毒對(duì)血液和與免疫相關(guān)的組織器官表現(xiàn)出更高的親和性，并且在3．5天內(nèi)開(kāi)始出現(xiàn)體外排毒；CSFVRTPCR和免疫組化兩種方法在急性感染模型的檢測(cè)中具有較高的符合性，符合率為86．36％。上述研究對(duì)CSFV臨床診斷具有重要的指導(dǎo)意義，臨床急性感染豬應(yīng)首先采集血液和免疫器官，其次為空腸、回腸段等組織器官。但是在對(duì)感染豬只進(jìn)行處理時(shí)必須嚴(yán)格控制對(duì)感染豬胴體的處理，以防止疾病擴(kuò)散。運(yùn)用本研究所建立的CSFVRTPCR對(duì)我國(guó)18省市644份疑似豬瘟樣品進(jìn)行檢測(cè)，再將檢測(cè)為陽(yáng)性的樣品序列進(jìn)行分析，以研究國(guó)內(nèi)豬瘟流行態(tài)勢(shì)和地理分布。結(jié)果644份疑似樣品中檢出CSFV陽(yáng)性74份，測(cè)回序列73條經(jīng)過(guò)DNASTAR分析，發(fā)現(xiàn)73株流行株分別為基因1．150．68％、2．142．47％、2．25．48％等3個(gè)亞群，與HCLV核苷酸同源性在75．7％100％之間與SM核苷酸同源性在74．3％一10006之間，流行株推導(dǎo)氨基酸序列與HCLV和SM株氨基酸序列同源性均在85．5％一100％之間；73株野毒以“北京、天津、河北湖北廣東”軸東西發(fā)散式分布在我國(guó)中東部地區(qū)；研究顯示目前無(wú)基因3群流行株存在。本研究成功建立了CSFVRTPCR檢測(cè)方法，為CSFV的臨床診斷提供了重要的技術(shù)支持，臨床應(yīng)用顯示本研究所建立的方法也可以用于CSF流行態(tài)勢(shì)的監(jiān)測(cè)；同時(shí)通過(guò)對(duì)急性感染的研究得到人工感染模型中病毒的組織分布和排毒規(guī)律，為進(jìn)一步研究CSFV的致病機(jī)理增添了新的數(shù)據(jù)。關(guān)鍵詞豬瘟RTPCR；免疫組化；分布；遺傳進(jìn)化II

簡(jiǎn)介：東北農(nóng)業(yè)大學(xué)碩士學(xué)位論文共表達(dá)豬細(xì)小病毒VP2與大腸桿菌不耐熱腸毒素B亞單位重組乳酸桿菌構(gòu)建及其免疫學(xué)初步評(píng)價(jià)姓名王相清申請(qǐng)學(xué)位級(jí)別碩士專業(yè)預(yù)防獸醫(yī)學(xué)指導(dǎo)教師李一經(jīng)20090620東北農(nóng)業(yè)大學(xué)農(nóng)學(xué)碩士學(xué)位論文II作為一種安全有效的佐劑，具有極大的應(yīng)用潛力。共表達(dá)VP2LTB重組干酪乳桿菌免疫組與單獨(dú)表達(dá)VP2重組干酪乳桿菌免疫組相比，PPG1VP2LTBLCASEI393或PPG2VP2LTBLCASEI393中小鼠血清中IGG的效價(jià)高于PPG1VP2LCASEI393或PPG2VP2LCASEI393組，與免疫對(duì)照組比較，差異顯著，并且分泌型的重組菌免疫性更好。在本研究中，口服免疫表達(dá)VP2LTB的重組乳酸菌不僅能誘導(dǎo)產(chǎn)生黏膜免疫，而且誘導(dǎo)產(chǎn)生了系統(tǒng)免疫，并且通過(guò)口服免疫產(chǎn)生的黏膜免疫反應(yīng)不只局限于胃腸道，也引起了其他黏膜部位的免疫反應(yīng)，而且共表達(dá)VP2LTB組的重組乳酸菌免疫后的SIGA抗體水平高于單獨(dú)表達(dá)VP2組的重組乳酸菌。機(jī)體通過(guò)分泌性IGA抗體在黏膜表面對(duì)病原體進(jìn)行排斥和清除，這對(duì)預(yù)防細(xì)小病毒感染是至關(guān)重要的。中和試驗(yàn)結(jié)果表明，PPG1VP2LCASEI393組、PPG2VP2LCASEI393組、PPG1VP2LTBLCASEI393組、PPG2VP2LTBLCASEI393組在免疫后49D血清的抗體中和效價(jià)分別為1304、1317、1338、1352。本研究首次構(gòu)建了重組大腸桿菌不耐熱腸毒素B亞單位和豬細(xì)小病毒主要免疫保護(hù)性抗原VP2蛋白的干酪乳桿菌細(xì)胞表面表達(dá)和分泌表達(dá)系統(tǒng)。本研究所獲得的結(jié)果表明，干酪乳桿菌可用做口服免疫傳遞抗原引起黏膜和系統(tǒng)免疫。當(dāng)VP2蛋白與LTB共表達(dá)時(shí)，加強(qiáng)了黏膜免疫反應(yīng)，LTB具有較好的免疫原性、抗原性和佐劑特性，可被用于基因工程疫苗的研制，很適合于用做黏膜疫苗的免疫佐劑。本研究為進(jìn)一步研究新型、有效的豬細(xì)小病毒口服疫苗奠定了基礎(chǔ)。關(guān)鍵詞關(guān)鍵詞豬細(xì)小病毒VP2蛋白；干酪乳桿菌；大腸桿菌不耐熱腸毒素B亞單位；共表達(dá)；免疫學(xué)評(píng)價(jià)

簡(jiǎn)介：中國(guó)農(nóng)業(yè)科學(xué)院碩士學(xué)位論文小反芻獸疫病毒抗體、抗原免疫學(xué)檢測(cè)方法和納米金核酸檢測(cè)方法的建立姓名邱文英申請(qǐng)學(xué)位級(jí)別碩士專業(yè)預(yù)防獸醫(yī)學(xué)指導(dǎo)教師李剛201106ABSTRACTPESTEDESPETITSRUMINANTSPPIT，ISAHIGLYCONTAGIOUSVIRALDISEASEAFFECTINGDOMESTICANDWILDSMALLRUMINANTSINRECENTYEARS，ITSPREADTONEWAREASINTHEWORLDPPRWAGFIRSTLYOUTBROKENINTIBETOFCHINAINJULY2007ITBECOMESMORENECESSARYANDCRITICALFORUSTEINFORCESTUDIESONDIAGNOSISANDCONTROLOFTHEDISEASEUSINGTHEPLASMIDWHICHCONTAINSTHECOMPLETEPPR、，NGENEANDFIVEPAIRSOFDESIGNEDPRIMERSACCORDINGTOTHECONSERVATISMOFNGENE，THEFIVESUBUNITSWEREAMPLIFIEDBYPCRTHENTHESUBUNITSWERECLONEDINTOPET32AEXPRESSIONVECTORSUCCESSFULLYTHESUBUNITSPROTEINWEREPURIFIEDTHERESULTSBYWESTERNBLOTSHOWEDTHATFOUROFTHESUBUNITSOFNPROTEINHADIMMUNOREACTIVITYWITHPPRVANTIBODIESONLYONEDIDNOTHAVETHERESULTSBYELISASHOWEDTHATTHEMAINBCELLEPITOPESSITEDONTHEFIFTHSUBUNITINOTHERWORDSSITEDONWZONETHEFIFTHSUBUNITPROTEINPETPPRVN5WHICHHADSTRONGIMMUNOREACTIVITYANDHIGHSPECIFICITYWASPURIFIEDANDUSEDASCOATINGANTIGENTODEVELOPEDAINDIRECTELISATODETECTTHEANTIBODIESAGAINSTPPRVTHEESTABLISHEDINDIRECTELISAWASSENSITIVEANDSPECIFICCOMPAREDWITHSTANDARDCOMPETITIVEELISA，THECOINCIDENCERATEBETWEENTWOMETHODSWAS98鐋TODEVELOPAGOLDIMMUNOCHROMATOGRAPHICASSAYGICAFORDETECTIONOFPPRVANTIGEN，POLYELONEANTIBODYWASPURIFIEDFROMHYPERIMMUNESERUMANDCONJUGATEDWITHCOLLOIDALGOLDPARTICLESPREPAREDBYTHETRISODIUMCITRATEREDUCTIONMETHODMEANWHILE，PURIFIEDRECOMBINANTPPRVNPROTEINANDTHEANTIGOATIGGWERESTRIPEDINTESTLINEPOSITIONANDINTHECONTROLLINEPOSITIONRESPECTIVELYTHEMINIMUMANTIGENDETECTEDBYDEVELOPEDGICAWAS10NGTHERESULTSSHOWEDTHATTHEGICAWASPECIFICANDEASYTOPERFORMITWASANACCEPTABLEALTERNATIVEFORTHENEINFIELDDIAGNOSISTODETECTMICROAMOUNTOFPPRVNUCLEOTIDEMOLECULES，AGOLDNANOPARTICLEBASEDMETHOD謝TLLSILVERSTAININGENHANCEMENTWASESTABLISHEDAPAIROFPROBESWHICHWERECOMPLEMENTARYTOTARGETNUCLEOTIDESWEREDESIGNEDANDLABELEDWITHBIOTINSANDNANOPATICLESRESPECTIVELYTHENTWOKINDSOFPROBESWEREHYBRIDIZEDWITHTARGETNUCLEOTIDES，THEHYBRIDIZATIONPRODUCTSWEREIMMOBILIZEDTOMICROPLATEVIAAVIDINBIOTINSYSTEM，ANDTHESIGNALSOFGOLDNANOPARTICLESWEREAMPLIFIEDWITHSILVERSTAININGSOLUTIONTHISMETHODCOULDDETECTASFEWAS1PMOL／LOFTARGETPPRVNUCLOTIDESINGENERAL，THREEMETHODSWEREESTABLISHEDTODETECTTHEANTIBODYANTIGENANDNUCLEOTIDESOFPPRⅥTHESEMETHODSWOULDBEUSEFULINTHEDETECTINGOFPPRVKEYWORDSPPRVSUBCLONE；INDIRECTELISA；GOLDIMMUNOCHOMATOGRAPHICASSAY；GOLDNANOPATICLEBASEDPROBEIII

簡(jiǎn)介：THEIDENTIFICATIONANDMOLECMLARBIOLOGICALCHARACTEIUSTICSANALYSESOFJA剛ESEENCEPHAI。ITISVIRUSISOLATEDFROM2008TO2009BYMAKUNSUPER訂SORPROFCHENPUYANADISSERTATIONSUBMITTEDTONANJINGAGRICULTURALUNIVERSITYINPARTIALFUMLLMENT0FTHEREQUIREMENTFORTHEMASTERDEGREEOFAGRONOMYVBTERINARYMEDICINECOMPLETEDINNOVEMBER，2009COMMENCEMENTINDECEMBER，2009原創(chuàng)性聲明本人鄭重聲明所呈交的學(xué)位論文，是本人在導(dǎo)師的指導(dǎo)下，獨(dú)立進(jìn)行研究工作所取得的成果。除文中已經(jīng)注明引用的內(nèi)容外，本論文不包含任何其他個(gè)人或集體已經(jīng)發(fā)表或撰寫過(guò)的作品成果。對(duì)本文的研究做出重要貢獻(xiàn)的個(gè)人和集體，均已在文中以明確方式標(biāo)明。本人完全意識(shí)到本聲明的法律結(jié)果由本人承擔(dān)。學(xué)位論文作者需親筆簽名溯7年肜月7日學(xué)位論文版權(quán)使用授權(quán)書本學(xué)位論文作者完全了解學(xué)校有關(guān)保留、使用學(xué)位論文的規(guī)定，同意學(xué)校保留并向國(guó)家有關(guān)部門或機(jī)構(gòu)送交論文的復(fù)印件和電子版，允許論文被查閱和借閱。本人授權(quán)南京農(nóng)業(yè)大學(xué)可以將本學(xué)位論文的全部或部分內(nèi)容編入有關(guān)數(shù)據(jù)庫(kù)進(jìn)行檢索，可以采用影印、縮印或掃描等復(fù)制手段保存和匯編學(xué)位論文。／保密口，在年解密后適用本授權(quán)書。本學(xué)位論文屬于不保密口。請(qǐng)?jiān)谝陨戏娇騼?nèi)打“√“學(xué)位論文作者需親筆簽名別滁射銘燃日日戶，D，●月月2≯年年冉77仃陽(yáng)如加