摘要
根据牛病毒性腹泻病毒（ＢＶＤＶ）ＮＡＤＬ株和猪瘟病毒（ＨＣＶ）Ａｌｆｏｒｔ株的核苷酸序列，设计合成了１对ＢＶＤＶ引物和３对ＨＣＶ引物。以从吉林、长春、哲盟３个地区经临床及病理学诊断为猪瘟的病料中提取的ＲＮＡ为模板，采用反转录－聚合酶链反应（ＲＴ－ＰＣＲ），分别以ＢＶＤＶ和ＨＣＶ引物进行扩增。结果，用ＢＶＤＶ引物从哲盟地区的疑似猪瘟病料中扩增出大小约４００ｂｐ的片段，而用３对ＨＣＶ引物在不同的条件下均未从该病料中扩增出相应大小的ＨＣＶ基因片段。此外，用ＨＣＶ的ＰＥ０／ＰＥ４引物从吉林、长春的病料中扩增出了ＨＣＶ的基因片段，但用ＢＶＤＶ引物从这两个地区的病料中均未扩增出ＢＶＤＶ的基因片段。从哲盟疑似猪瘟病料中扩增出的片段经克隆、序列测定及计算机分析证实，该片段的核苷酸序列和氨基酸序列与ＢＶＤＶ的同源性明显高于与ＨＣＶ的同源性。将哲盟病料接种于ＭＤＢＫ传代细胞，出现典型而规律的ＢＶＤＶ样细胞病变。由此证明。
Based on the published sequences of bovine viral diarrhea virus (BVDV) strain NADL and hog cholera virus (HCV) strain Alfort, one set of BVDV specific primers and 3 sets of HCV specific primers were designed, chemically synthesized and used to amplify the cDNA sequences of corresponding viruses possibly existing in spleen tissue samples of suspicious HC cases from Jilin, Changchun and Zhemeng regions by reverse transcription polymerase chain reaction (RT PCR). The results showed that a fragment with a size of 400 bp was successfully amplified from the tissue sample of the suspect HC cases from Zhemeng with BVDV primers, but no respective fragments were amplified with 3 sets of HCV primers under selected conditions. However, from the tissue samples of suspicious HC cases of Jilin and Changchun regions, the expected fragments were obtained easily with HCV primers, whereas no fragment were amplified with BVDV primers. Comparison of the nucleotide and amino acid sequences of the fragment amplified with BVDV primers with those of BVDV strains and HCV strains showed that the homologies of nucleotide and amino acid sequences with BVDV are much higher than those of HCV, indicating the corresponding virus is a BVDV, named 'ZM 95'. This BVDV strain exhibited typical and regular BVDV cytopathic effect (CPE) in MDBK cell culture. This is the first report in China that BVDV can naturally infect pigs and may cause a disease resembling hog cholera.
出处
《中国兽医学报》
CAS
CSCD
1996年第4期341-345,共5页
Chinese Journal of Veterinary Science
基金
国家自然科学基金
关键词
猪瘟
牛
腹泻病毒
聚合酶链反应
HC
BVDV
reverse transcription polymerase chain reaction (RT PCR)
