摘要
以3 2 P标记犬冠状病毒特异性RT_PCR产物制成核酸探针 ,与CCVNL_18株、犬瘟热病毒、犬细小病毒、犬副流感病毒、犬腺病毒、狂犬病病毒反转录产物、正常CRFK细胞以及做 10～ 10 0 0 0倍稀释的CCVNL_18株反转录产物杂交 ,进行核酸探针的敏感性和特异性试验。结果表明 ,该探针仅与CCVNL_18株反转录产物呈阳性杂交 ,与对照病毒和正常细胞反转录产物均呈阴性反应。初步应用试验结果 ,该探针可与国内CCV分离病毒YS1、CI1株杂交 ,且可从 8份腹泻犬粪便中检出 5份CCV阳性病料。本研究为在我国开展犬冠状病毒感染的分子流行病学调查和临床诊断提供了一种敏感。
Partial spike protein gene of Canine coronavirus amplified with the method of RT_PCR was radiolabelled with 32 P to develop the nucleotide probe. Specificity and sensitivity tests were carried out with the nucleotide probe to detect the reverse transcription (RT) products of CCV NL_18 strain, canine distemper virus, canine parvovirus, canine parainfluenza virus, rabies virus, CRFK cell without incubating virus and 10 1～10 4 dilution of RT products of CCV NL_18 strain. The results showed that the probe could only hybridized with the RT product of CCV NL_18 strain but not with that of control viruses and CRFK cell. And it could detect 1 μL 1×10 3 dilution of RT product of CCV NL_18 strain. The results of primary application indicated that the probe could hybridized the RT products of CCV YS1, CI1 strain isolated in china and could detect 5 CCV positive samples out of 8 dogs from the diahorrae. The results laid foundation for the molecular epidemiology and clinical diagnosis of CCV infection.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2003年第3期171-174,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
"九五"军队医药卫生重点项目编号 98Z0 35
关键词
犬冠状病毒
核酸探针
初步应用
canine coronavirus
nucleotide probe
primary application
