重庆地区动物BDV感染的分子生物学研究

发布时间：2018-06-20 06:09

本文选题：博尔纳病病毒 + 感染　；参考：《重庆医科大学》2007年硕士论文

【摘要】： 第一部分重庆地区家兔博尔纳病病毒自然感染状况研究目的:检测重庆地区家兔博尔纳病病毒(Borna Disease Virus ,BDV)隐性带毒情况。分析该BDV的可能种系来源及其与标准株strain V/FR、strain He/80/FR和strain H1766的同源性。方法:采用巢式逆转录酶PCR结合荧光定量PCR(Real-time nRT-PCR)检测重庆地区60例健康家兔脑组织BDV P24基因片段,并对其中的阳性产物进行基因序列测定,然后用BLAST软件以及DNAstar V6.13 EditSeq软件对测序结果进行分析。结果:2例家兔为阳性。该BDV P24片段的核苷酸序列与马源BDV strain H1766株同源性最高,为97.67%,与标准株strain V/FR和strain He/80/FR同源性均为96.51%,但是它们编码的氨基酸相同。结论:重庆地区家兔中存在动物源性BDV隐性感染,该BDV P24核苷酸序列与strain V/FR和strain He/80/FR具有高度同源性。但是与马源的strain H1766同源性最高。第二部分重庆地区狗博尔纳病病毒自然感染状况的初步探讨目的:初步了解重庆地区狗BDV的隐性感染情况,对检测到的阳性片段进行核苷酸序列比对,探讨其可能的种系来源及其与国外公认的标准株strain V/FR、strain He/80/FR和strain H1766的同源性。方法:采用Real-time nRT-PCR检测重庆地区40例健康狗脑组织(右侧前额叶)BDV P24基因片段,并对其中的阳性产物进行基因序列测定,然后用BLAST软件以及DNAstar V6.13 EditSeq软件对测序结果进行分析。结果:5例狗为阳性。该BDV P24片段的核苷酸序列与标准株strain H1766和strain V/FR同源性最高,为97.67%。与strain He/80/FR的同源性为94.19%,与已有报道的奥地利狗源BDV同源性为95.35%。但是它们编码的氨基酸相同。结论:重庆地区狗中存在动物源性BDV的隐性感染,该BDV P24核苷酸序列与strain H1766、strain V/FR和strain He/80/FR具有高度同源性。第三部分重庆地区鸽子博尔纳病病毒自然感染状况的研究目的:初步了解重庆地区鸽子博尔纳病病毒(Borna DiseaseVirus,BDV)隐性感染状况,对其感染的病毒株进行核苷酸序列比对,探讨其可能的种系来源以及与国外公认的标准株strain V/FR、strain He/80/FR和strain H1766的同源性方法:本研究采用Real-time nRT-PCR技术对重庆地区60只鸽子的外周血单核细胞(PBMCs)中的BDV P24基因片段进行了检测,并且对阳性PCR产物进行测序。然后用BLAST软件以及DNAstar V6.13 EditSeq软件对该片段进行核苷酸比对及序列分析。结果:四例样本为阳性。该BDV P24片段的核苷酸序列与法国狐狸分离的BDV同源性最高,为97.67%。与标准株strain V/FR和strain He/80/FR的同源性都是96.51%,与strain H1766同源性仅为95.35%。但是它们编码的氨基酸相同。结论:本研究表明重庆地区鸽子存在博尔纳病病毒的隐性感染,其感染病毒的P24核苷酸片段与strain V/FR、strain He/80/FR以及strain H1766等国际公认的标准病毒株存在高度同源性。变异程度均小于4.65%,且其编码的氨基酸无差别。
[Abstract]:A study on the natural infection status of Borna disease virus in the first part of Chongqing

Objective : To investigate the recessive herpes zoster virus ( BDV ) in rabbits with Borna Disease Virus ( BDV ) . The possible source of BDV and its homology with strain V / FR , strain He / 80 / FR and strain H1766 were analyzed .

Methods : The BDV P24 gene fragment of 60 healthy rabbits was detected by nested reverse transcriptase polymerase chain reaction ( Real - time nRT - PCR ) , and the positive products were sequenced . Then the sequencing results were analyzed by BLAST software and DNAstar V6 . 13 EditSeq software .

Results : The homology of nucleotide sequence of BDV P24 fragment was 97.67 % , which was 96.51 % and that of strain V / FR and strain He / 80 / FR was 96.51 % , but the amino acid sequence was the same as that of the standard strain V / FR and strain He / 80 / FR .

Conclusion : The BDV P24 nucleotide sequence is highly homologous to strain V / FR and strain He / 80 / FR in the rabbit of Chongqing . However , it has the highest homology with strain H1766 .

A Preliminary Study on the Natural Infection of Dog Borna Disease Virus in the Second Part of Chongqing

Objective : To investigate the recessive infection of BDV in Chongqing , and to investigate the nucleotide sequence alignment of the detected positive fragment , and explore its possible source of infection and its homology with other internationally recognized strain V / FR , strain He / 80 / FR and strain H1766 .

Methods : The BDV P24 gene fragment was detected by Real - time nRT - PCR in 40 healthy dog brain tissues ( right anterior frontal lobe ) , and the positive products were sequenced . Then the sequencing results were analyzed by BLAST software and DNAstar V6 . 13 EditSeq software .

Results : 5 cases of dogs were positive . The nucleotide sequence of BDV P24 fragment was the highest with strain H1766 and strain V / FR of strain H1766 and strain V / FR , which was 97.67 % .

Conclusion : The BDV P24 nucleotide sequence has high homology with strain H1766 , strain V / FR and strain He / 80 / FR .

A study on the natural infection status of pigeon Borna disease virus in the third part of Chongqing

Objective : To investigate the recessive infection status of Borna DiseaseVirus ( BDV ) in Chongqing region , to investigate the nucleotide sequence alignment of the infected virus strains , to explore the possible source of infection and to share the homology of strain V / FR , strain He / 80 / FR and strain H1766 .

Methods : The BDV P24 gene fragment in peripheral blood mononuclear cells ( PBMCs ) of 60 pigeons in Chongqing was detected by Real - time nRT - PCR , and the positive PCR products were sequenced . Then , the nucleotide alignment and sequence analysis were performed with BLAST software and DNAstar VB.13 EditSeq software .

Results : Four samples were positive . The nucleotide sequence of the BDV P24 fragment was up to 97.67 % . The homology with strain V / FR and strain He / 80 / FR was 96.51 % . The homology with strain H1766 was only 95.35 % , but the amino acids were the same .

Conclusion : This study shows that there is a recessive infection of Borna disease virus in the pigeon in Chongqing . The P24 nucleotide fragment of its infected virus has high homology with strain V / FR , strain He / 80 / FR and strain H1766 . The degree of variation is less than 4.65 % , and its encoded amino acid is no difference .
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R373

【引证文献】