果子狸细小病毒的分离与鉴定

Isolation and Identification of Parvovirus in Paguma larvata

用F81细胞从河南送检的患有肠炎的果子狸粪便样品中分离出一株病毒,经形态学、理化学、生物学和分子病毒学实验鉴定是一株CPV-2a的突变株。此毒株在F81细胞上生长,能产生CPV感染的特征性细胞病变,细胞肿胀、变圆、破碎、脱落;病毒粒子二十面体立体对称,直径为20～24nm;无囊膜,能抵抗氯仿的处理,耐酸,耐热,但能被5-IUDR抑制。可凝集猪红细胞,不凝集鸡、豚鼠、大鼠、兔、人"O"型红细胞,能被抗CPV抗体所抑制。设计特异性细小病毒引物扩增VP2基因的两个片段,并进行测序,分析结果表明,该毒株是在CPV-2a的基础上其VP2的第297位氨基酸发生A→S突变,第300位氨基酸发生G→S突变,第301位氨基酸发生T→A突变。该毒株暂定名为PV GZL HN 1 02。该病毒能感染犬。

A Paguma larvata Parvovirus strain was isolated from the feces.It was designated as a mutant of CPV-2a through a series of systematic identification assays, such as morphological, physicochemical, biological and molecular virology assay. It can grow in F81 cell line and produce cytopathic effects with the characteristics of CPV, swelling, rounding, broken, exfoliation. The virion shows icosachedral symmetry, 20～24nm in diameter. The strain resists the treatment of chloroform and is stable in acid and hot environment, but is suppressed by 5-IUDR.The virus can agglutinate RBC of pig, but not of human, chicken,rabbit,rat,guinea pig.The positive antibody against CPV can inhibit HA of it.The results of PCR products sequence including two fractions of carnivore parvovirus VP2 gene show it possessed a A→S substitution at amino acid residue 297 ,a G→S substitution at amino acid residue 300,a T→A substitution at amino acid residue 301 based on VP2 amino acid sequences of conventional CPV-2a.It is temporarily named as PV/ Paguma larvata /1/02. The result of animal testing showed that it could infect dogs.