摘要
根据猪细小病毒(PPV)基因组序列,设计引物克隆了PPV YK株保守序列NS1基因,将NS1基因纯化,利用地高辛标记制备了NS1基因PCR产物探针和克隆NS1探针,两种核酸探针的标记都达到了0.1 pg/μL。特异性试验结果表明,这两种探针都能与PPV DNA发生特异的阳性杂交,而与对照的PRV、PCV、PRRSV和HCV的核酸杂交呈阴性。敏感性试验结果表明,克隆NS1探针敏感度为0.5 pg/μL,NS1基因PCR产物探针敏感度为1 pg/μL。综上所述,这两种DIG标记探针特异性强,敏感性高,可用于PPV的检测。
According to the genomic sequences of PPV a pair of primers were designed, and the highly conservative NS1 gene of PPV YK strain was amplified by PCR. Then the purified NS1 PCR products and clone of NS1 gene were labeled with digoxigenin as DNA probe for PPV,the 0.1 pg/μL of the labeling efficiency of two probes was obtained. The results of specificity assay of hybridization showed that PPV DNA was positive,but other control nucleotide extracted from PRV,PCV, PRRSV and HCV were negative. The resuits of sensitivity assay of hybridization exhibited that 0. 5 pg of PPV DNA could be detected by NS1 cloned probe,and 1 pg of PPV DNA could be detected by NS1 PCR products probe. In summary,the two digoxigenin-labeled probes are of high specificity and sensitivity,and could be used to detect the PPV.
出处
《动物医学进展》
CSCD
2005年第9期80-84,共5页
Progress In Veterinary Medicine
关键词
猪细小病毒
地高辛标记核酸探针
杂交
Porcine parvovirus
the digoxigenin-labeled probes hybridization
