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应用套式PCR在MDCK细胞系中发现犬细小病毒

Presence of a Vertically Transmitted Canine Parvovirus in MDCK Cell Line Revealed with a Nested PCR

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【作者】 杨德威宋延华刘福安

【Author】 YANG De wei(YEUNG De wei), SONG Yan hua, LIU Fu an (Dept. of Veterinary Medicine, South China Agric. Univ., Guangzhou 510642, China)

【机构】 华南农业大学动物医学系!广东广州510642

【摘要】 本研究自 1 997年至 1 999年从国内部分大学实验室、卫生防疫站和动检局陆续收集 1 1个犬肾 (MDCK)细胞系样品 ,选取犬细小病毒基因组的VP2 基因上 4段核苷酸序列作引物 ,对样品DNA进行套式PCR(NestedPCR)检测 ;PCR扩增产物经 0 0 1g/mL琼脂糖凝胶电泳和克隆到pGEM-T载体并进行核苷酸序列测序鉴定后 ,发现我国MDCK细胞系中存在犬细小病毒的传代病毒株 该病毒基因组部分序列测定结果显示与犬细小病毒CPV N株有 91 %同源性 .

【Abstract】 Eleven samples of MDCK cell lines collected from various university laboratories, epidemic prevention stations and quarantine bureaus within China since 1997. Utilizing 4 stretches of nucleotide sequences reported for the VP 2 gene of canine parvovirus as primers, the DNA of the samples were screened with a nested PCR procedure. The PCR amplified product was subjected to electrophoretic analysis in 0.01 g/mL agarose, cloned in a PGEM T vector and the nucleotide sequence determined. The findings revealed the presence of a vertically transmitted canine parvovirus. Partial sequencing of the virus genome showed 91% nucleotide homology with the canine parvovirus, strain CPV N. This is the first case report of canine parvovirus in MDCK cell lines in our country.

【关键词】 犬细小病毒MDCK细胞套式PCR
【Key words】 canine parvovirusMDCK cell linenested PCR
  • 【文献出处】 华南农业大学学报 ,JOURNAL OF SOUTH CHINA AGRICULTURAL UNIVERSITY , 编辑部邮箱 ,2000年03期
  • 【分类号】S852.655;S85265
  • 【被引频次】23
  • 【下载频次】187
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