摘要
将疑似断奶仔猪多系统衰竭综合征（PMWS）病猪肺脏研磨、过滤除菌后接种无PCVl污染的PKl5细胞，盲传4代，用PCR和免疫过氧化物酶单层试验（IPMA）检测。同时，利用所合成的PCV2全长的特异性引物进行病毒基因组全长的扩增及克隆、测序与比对分析。结果显示，不同传代细胞中PCV2核酸均为阳性，且PCVl特异引物检测为阴性，表明分离到的PCV2无PCVl污染，将该毒株命名为PCV2B1株。序列分析表明，该病毒基因组全长1767lap，与其他毒株的核苷酸同源性在95．4％～99．8％之间，ORFl的氨基酸的同源性在98．4％～99．7％之间，ORF2的氨基酸的同源性在89．7％～100％，核苷酸和氨基酸的序列与保定株BDlA（DQ910865）的序列最接近。
The lung from the pig with suspected PMWS was homogenized and filtered. The suspension was inoculated in PK15 cells free of PCV1 and cultivated for 4 passages. And porcine circovirus type 2（PCV2） and PCV1 in each passage was detected by PCR with specific primers and IPMA, respectively. Meanwhile, the full length of viral genome was amplified and cloned with a pair of specific primers by PCR, and the recombinant plasmid was sequenced and analyzed. The results showed that the detection of PCV2 was positive, but the detection of PCV1 was negative in each passage, suggesting that the isolated PCV2 strain was not contaminated with PCV1. This isolated strain was named as PCV2 B1 strain. Sequence analysis showed that its complete genome was composed of 1767 nucleotides. The homology of nucleotide sequence in PCV2 B1 strain was 95.4~99.8% compared with that of others, and the homology of ORF1 and ORF2 amino acid were 98.4-99.7% and 89.7-100%,respectively. The sequence of nucleotide and amino acid of PCV2 B1 was very similar to that of BD1A （DQ910865）.
出处
《北京农学院学报》
2007年第3期20-24,共5页
Journal of Beijing University of Agriculture
基金
北京市自然科学基金资助项目(6062006)
北京市优秀人才资助项目(20061D0502100282)
