Zhang Chuanmei, Yu Yongle, Yang Haiyan, Li Guimei, Yu Zekun, Zhang Hongliang, Shan Hu
Shandong Key Laboratory of Preventive Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China.
Shandong Key Laboratory of Preventive Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China.
J Virol Methods. 2014 Dec 15;210:1-6. doi: 10.1016/j.jviromet.2014.09.014. Epub 2014 Sep 26.
A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay has been developed to detect and differentiate between canine parvovirus (CPV) and mink enteritis virus (MEV). Eight CPV and three MEV epidemic strains isolated from 28 pathological samples from dogs and minks suspected of being infected with parvovirus were amplified by PCR using a pair of specific primers designed based on the CPV-N strain (M19296). PCR amplified a fragment of 1016bp from the genomic DNA of both MEV and CPV. The MEV-derived fragment could be digested with the restriction enzyme BSP1407I into three fragments of 102bp, 312bp and 602bp, while the fragment amplified from the CPV genomic DNA was digested into only two fragments of 414bp and 602bp. The lowest DNA concentration of CPV and MEV that could be detected using this assay was 0.004μg/ml and 0.03μg/ml, respectively. The PCR-RFLP assay developed in the present study can, therefore, be used to detect and differentiate MEV from CPV with high specificity and sensitivity.
已开发出一种聚合酶链反应-限制性片段长度多态性（PCR-RFLP）检测方法，用于检测犬细小病毒（CPV）和水貂肠炎病毒（MEV）并进行区分。从28份疑似感染细小病毒的犬和水貂的病理样本中分离出8株CPV和3株MEV流行株，使用基于CPV-N株（M19296）设计的一对特异性引物，通过PCR进行扩增。PCR从MEV和CPV的基因组DNA中扩增出一个1016bp的片段。MEV来源的片段可被限制性内切酶BSP1407I消化为102bp、312bp和602bp的三个片段，而从CPV基因组DNA扩增出的片段仅被消化为414bp和602bp的两个片段。使用该检测方法可检测到的CPV和MEV的最低DNA浓度分别为0.004μg/ml和0.03μg/ml。因此，本研究开发的PCR-RFLP检测方法可用于高特异性和高灵敏度地检测MEV并将其与CPV区分开来。
