Isolation of egg-laying reduction syndrome virus from breeder Muscovy duck and its polymerase chain reaction identification
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摘要: 从临床表现产蛋率下降、产蛋异常的种番鸭输卵管粘膜中分离到1株病毒,经血凝及血凝抑制试验鉴定为产蛋下降综合征病毒,命名为E05。参照GenBank上发表的鸭源产蛋下降综合征病毒基因组序列的保守区设计1对引物,通过PCR技术扩增出286 bp的基因片段,将扩增产物与T载体连接后,转化感受态细胞,筛选出阳性重组质粒进行测序分析。测序结果与GenBank上已发表的EDSV 100kD蛋白序列比对,扩增片段与已登陆的基因片段完全相符,同源性100%。Abstract: A virus strain,E05,was isolated from oviduct mucosa of breeder Muscovy duck,which showed irregularity and reduction in egg-laying.The virus was identified as an egg-laying reduction syndrome virus(EDSV) by haemagglutination and haemagglutination inhibition tests.According to the conserved region of duck original EDSV sequence,a pair of primers was designed for PCR.The product of the PCR was ligated with T vector and introduced into competent cells.Positive recombinant was selected for sequencing.The sequence of PCR product was compared with that of the 100 kD gene found in GenBank nucleotide database.The result showed that the amplified product from the isolated virus was completely identical with the counterpart of the EDSV.
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Key words:
- breeder Muscovy duck /
- egg-laying reduction syndrome virus /
- isolation /
- identification
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