鹅源副黏病毒的分离鉴定及部分特性研究

Isolation and Identification of the Goose Paramyxovirus Strain JG04 and Study on Its Partial Characters

采用9日龄鸡胚,从长春某个体养鹅户的病死鹅的肝、脾和肾脏中分离到1株病毒,暂定名为JG04株。经HA和HI试验、电镜观察、常规RT-PCR、NDV荧光RT-PCR检测,证实JG04株为鹅源副黏病毒。与GenBank下载的9株NDV毒株的HN基因作同源性比较,并进一步对JG04株进行耐酸试验、氯仿和乙醚敏感试验、血凝解脱及血凝素热稳定性试验、EID50、ELD50和毒力测定。结果表明:JG04株与江苏近年来分离的鹅源副黏病毒株的HN基因同源性较高,核苷酸同源性高达96.4%,氨基酸同源性高达96.9%;JG04株有良好的血凝性,能凝集人和鸡的红细胞;电镜观察病毒粒子有囊膜,直径约200 nm;对酸、氯仿、乙醚敏感;血凝解脱为快速解脱型,血凝素热稳定性差;EID50、ELD50分别为1.74×109/mL、2.57×109/mL;MDT为54.2 h,ICPI为1.66,IVPI为2.60。上述结果表明,GJ04株与新城疫病毒(NDV)速发型具有类似的毒力,属强毒力毒株。

Using chicken embryos, one strain of virus named JG04 was isolated from the liver, spleen and kidney of a dead goose in Changchun. This isolate was identified paramyxovirus by hemagglutination （HA） and hemagglutination inhibition（HI）, electron micrograph （EM）, RT-PCR, NDV fluorescent RT- PCR. Then its characters of resistance to acid environment, sensitivity to chloroform and diethyl ether, hemagglutinating free, hemagglutining stability, chicken embryo＇s median infectious dose（EIDso）, chicken embryo＇s minimum lethal dose（ELDso）, mean death time（MDT） to chicken embryo, intracerebral pathogenicity index （ICPI） to 1-day-old chicken and intravenous pathogenicity index （IVPI）to 6-week-old chicken were studied. It was found that the virus was able to agglutinate chicken and human erythrocytes and inhibited by serum against newcastle disease virus（NDV）, but not inhibited by serum against EDS- 76,AIV H5 and H9 subtype. The isolate possessed envelope and about 200nm in diameter observed in negatively-stained transmission electron micrograph. The EIDso, ELDso, MDT, ICPI, IVPI of the virus were 1.74 ×10^9/mL, 2.57 ×10^9/mL, 54.2 h, 1.66 and 2.60 respectively. These results suggested the JG04 strain belonged to velogenic strain.