【摘要】：2012-2014年從湖南平江、淚羅、瀏陽、長(zhǎng)沙四地的四個(gè)規(guī)模豬場(chǎng)送檢的病料(腦組織)中檢測(cè)到偽狂犬病毒(PRV)野毒。將腦組織接種PK-15細(xì)胞,經(jīng)PCR和動(dòng)物接種鑒定為偽狂犬病毒。4個(gè)毒株分別命名為PRV-XiangA、PRV-GA、PRV-YY和PRV-LY。本研究通過對(duì)這4株P(guān)RV的免疫(gB、gG、gH、gI、gL、gM)與毒力相關(guān)基因(gE、PK、TK)進(jìn)行序列分析,結(jié)果顯示這4株P(guān)RV的免疫與毒力相關(guān)基因核苷酸同源性在99.8%-100%,說明分離得到的4株P(guān)RV變異極小；將這4株P(guān)RV與國內(nèi)外的主要毒株進(jìn)行序列比對(duì),結(jié)果表明這4株P(guān)RV與國內(nèi)的分離毒株同源性很高,與BJ-YT株和TJ株的親緣關(guān)系最近,為99.8%-100%,說明當(dāng)前國內(nèi)流行的PRV毒株變異較小。對(duì)從湖南省分離得到的4株偽狂犬病毒進(jìn)行毒價(jià)測(cè)定后,篩選出一株培養(yǎng)滴度較高的偽狂犬病毒,即PRV-YY株,該株偽狂犬病毒在MDBK細(xì)胞上的病毒滴度能夠達(dá)到107.41 TCID50/0.1mL。通過建立MDBK單克隆細(xì)胞庫,篩選出1株單克隆細(xì)胞MDBK-1培養(yǎng)PRV-YY株的病毒滴度能夠達(dá)到107.71 TCID50/0.1mL,通過終點(diǎn)稀釋法將PRV-YY株在MDBK-1上進(jìn)行傳代,PRV-YY株在該株單克隆細(xì)胞MDBK-1上的病毒滴度能夠穩(wěn)定在107.5TCID50/0.1mL以上,PRV-YY株的平均病毒滴度達(dá)到107.692TCID50/0.1mL, PRV-YY株的病毒滴度在原有的基礎(chǔ)上提高了約2倍。這為制備偽狂犬病毒野外流行毒株滅活疫苗打下了良好的基礎(chǔ)。
[Abstract]:Pseudorabies virus (PRV) was detected from four pig farms in Pingjiang, Yiluo, Liuyang and Changsha from 2012 to 2014. The brain tissue was inoculated with PK-15 cells and identified as pseudorabies virus by PCR and animal inoculation. The four strains were named PRV-XiangAv GAPRV-YY and PRV-LY. In this study, the nucleotide homology of immune and virulence related genes (PRV) of the four strains of PRV were analyzed. The results showed that the nucleotide homology of the four strains of PRV was 99.8-100, which indicated that the PRV mutation of the four isolated strains was very small. The four PRV strains were sequenced with the main strains at home and abroad. The results showed that the four PRV strains had high homology with the domestic isolates, and had the closest relationship with BJ-YT and TJ strains (99.8-100), indicating that there was little variation in the current prevalent PRV strains in China. Four strains of pseudorabies virus isolated from Hunan Province were tested for virus titer. A pseudorabies virus strain, PRV-YY strain, was selected. The titer of pseudorabies virus on MDBK cells was 107.41 TCID50 / 0.1 mL. By establishing a MDBK monoclonal cell library, The virus titer of PRV-YY strain cultured with a monoclonal cell line MDBK-1 could reach 107.71 TCID 50 / 0.1 mL, and the virus titer of PRV-YY strain on MDBK-1 could be stabilized above 107.5TCID50/0.1mL by terminal dilution method. The virus titer of PRV-YY strain on MDBK-1 was stable at the end point dilution method. The average titer of PRV-YY strain was 107.692TCID 50 / 0.1mL, and the virus titer of PRV-YY strain increased by about 2 times. This lays a good foundation for the preparation of pseudorabies virus inactivated vaccine.
【學(xué)位授予單位】：湖南農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S852.65

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 占松鶴;劉華;周迎春;王軍;沈艷;何長(zhǎng)生;朱良強(qiáng);汪彤;范紅結(jié);;安徽省規(guī)模化種豬場(chǎng)豬偽狂犬病血清學(xué)調(diào)查[J];動(dòng)物醫(yī)學(xué)進(jìn)展;2015年03期

2 李元果;高曉龍;桑曉宇;王海軍;張成林;王愛榮;于志君;任志廣;鄭學(xué)星;馮娜;王鐵成;楊松濤;黃耕;高玉偉;夏咸柱;胡永浩;;虎源偽狂犬病毒的鑒定及gB基因分析[J];中國病原生物學(xué)雜志;2014年12期

3 趙鴻遠(yuǎn);彭金美;安同慶;李琳;冷超糧;陳家锃;王倩;常丹;張秋月;蔡雪輝;童光志;田志軍;;豬偽狂犬病病毒變異株的分離鑒定及其gE基因的分子特征[J];中國預(yù)防獸醫(yī)學(xué)報(bào);2014年07期

4 鐘承;張樂天;王巨實(shí);陸梓杰;劉蕾;楊萬蓮;孫艷爭(zhēng);呂艷麗;;犬源偽狂犬病毒BJ/RD株的分離鑒定及gE基因分析[J];中國動(dòng)物傳染病學(xué)報(bào);2013年05期

5 聶奎;李竹青;曾政;楊澤林;駱璐;;川渝部分地區(qū)育肥豬偽狂犬病血清流行病學(xué)調(diào)查及危險(xiǎn)性因素[J];西南大學(xué)學(xué)報(bào)(自然科學(xué)版);2013年09期

6 黃藝珠;張淵魁;魏建超;曾亮明;朱海俠;翁燕梅;傅星源;俞建萍;潘海城;謝海明;;豬偽狂犬病毒SD株的分離鑒定及TK基因序列分析[J];畜牧與獸醫(yī);2013年04期

7 楊睿;翟少欽;王孝友;沈克飛;鄭華;曾秀;;重慶地區(qū)豬偽狂犬野毒株感染的流行病學(xué)調(diào)查[J];黑龍江畜牧獸醫(yī);2013年01期

8 樊振華;姚敬明;孟帆;米瑞娟;羅甜甜;吳忻;劉文俊;王娟萍;;山西部分種豬場(chǎng)豬偽狂犬病分子流行病學(xué)調(diào)研[J];山西農(nóng)業(yè)科學(xué);2012年09期

9 張士強(qiáng);易悅;徐志文;朱玲;;豬偽狂犬病病毒SL株TK基因的克隆與生物信息學(xué)分析[J];中國畜牧獸醫(yī);2012年06期

10 邱美珍;傅勝才;杜麗飛;譚良溪;楊俊;李海輝;王慧;向衛(wèi)紅;金美中;周望平;;湖南省部分規(guī)模豬場(chǎng)偽狂犬病野毒血清流行病學(xué)系統(tǒng)監(jiān)測(cè)與凈化分析[J];養(yǎng)豬;2012年03期

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