应用套式PCR在MDCK细胞系中发现犬细小病毒
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Presence of a Vertically Transmitted Canine Parvovirus in MDCK Cell Line Revealed with a Nested PCR
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    本研究自1997年至1999年从国内部分大学实验室、卫生防疫站和动检局陆续收集11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(Nested PCR)检测;PCR扩增产物经0.01g/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.

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杨德威,宋延华,刘福安.应用套式PCR在MDCK细胞系中发现犬细小病毒[J].华南农业大学学报,2000,(3):81-83

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