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| 鹅细小病毒VP1-VP3非重叠序列地高辛探针的制备和应用 | |
| 引用本文： | 布日额,王君伟,吴金花,马波,Ulrich Neumann.鹅细小病毒VP1-VP3非重叠序列地高辛探针的制备和应用[J].中国兽医杂志,2005,41(1):7-10. |
| 作者姓名： | 布日额 王君伟 吴金花 马波 Ulrich Neumann |
| 作者单位： | 1. 东北农业大学动物医学院,黑龙江,哈尔滨,150030;内蒙古民族大学动物科技学院,内蒙古,通辽,028042 2. 东北农业大学动物医学院,黑龙江,哈尔滨,150030 3. 内蒙古民族大学动物科技学院,内蒙古,通辽,028042 4. 汉诺威兽医学院,德国,汉诺威,30559 |
| 基金项目： | 黑龙江省“十五”攻关课题 ( GB0 1B5 0 2 0 2 ) |
| 摘 要： | 根据GPV H1株核苷酸序列，设计了扩增VP1-VP3基因非重叠序列的1对引物，对其结构蛋白VP1与VP3非重叠核苷酸序列进行PCR扩增，将PCR产物纯化、回收后制备出GPV VP1-VP3基因DIG标记核酸探针，其标记效率达到0．1pg／μl。特异性检测结果表明，该探针能与GPV不同毒株核酸发生特异性杂交，而与对照的DPV、GPMV等病毒的核酸杂交反应均为阴性；敏感性检测结果表明该探针对GPV的最低检出量为0．032ng。上述试验结果表明该探针可以用于GPV感染临床病料的检测。 |
| 关 键 词： | VP1 地高辛 探针 VP3基因 特异性检测 阴性 临床 鹅细小病毒 病料 核苷酸序列 |
| 文章编号： | 0529-6005(2005)01-0007-04 |
Study on Digoxigenin-labelled nucleic acid probe for GPV VP1-VP3 nonrepeated DNA sequences |
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| BU Ri-e.Study on Digoxigenin-labelled nucleic acid probe for GPV VP1-VP3 nonrepeated DNA sequences[J].Chinese Journal of Veterinary Medicine,2005,41(1):7-10. | |
| Authors: | BU Ri-e |
| Abstract: | According to the gene sequences of GPV H1 strain designed a pair of primers and amplified the VP1-VP3 nonrepeated DNA fragment between VP1 and VP3 gene. Labeled VP1-VP3 DNA with Digoxigenin and prepared VP1-VP3 DNA probe for detection of GPV after purification and reception of the product of PCR. The result of specificity assay of hybridization showed that the DNA of the two strains of GPV were positive,and other nucleotide extracted from DPV, GPMV and allantoic fluid of the noninoculated embryonating eggs were negative. The result of sensitivity assay of hybridization showed that as little as 0.032 ng of GPV DNA could be detected by the DIG-labeled probe.According to the presnt positive results of this assay, this probe could hybridizate with GPV neucleotide which extracted from the clinical samples. So this DIG-labeled DNA probe could be used to detect the clinical samples of GPVI. |
| Keywords: | GPV VP1-VP3 gene nonrepeated sequence Digoxigenin neucleotide probe goose parvovirus |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |