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

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 .

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.