title: Development of a PCR-RFLP assay for the detection and differentiation of canine parvovirus and mink enteritis virus.
abstract: 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.
authors.affiliation: Shandong Key Laboratory of Preventive Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China.
authors.forename: Chuanmei
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Shandong Key Laboratory of Preventive Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China.
authors.forename: Yongle
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Shandong Key Laboratory of Preventive Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Yang
authors.affiliation: Shandong Key Laboratory of Preventive Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China.
authors.forename: Guimei
authors.initials: G
authors.lastname: Li
authors.affiliation: Shandong Key Laboratory of Preventive Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China.
authors.forename: Zekun
authors.initials: Z
authors.lastname: Yu
authors.affiliation: Shandong Key Laboratory of Preventive Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China.
authors.forename: Hongliang
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Shandong Key Laboratory of Preventive Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China. Electronic address: shanhu67@163.com.
authors.forename: Hu
authors.initials: H
authors.lastname: Shan
mesh_terms: D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004283:Dog Diseases / Q000821:virology*; D004285:Dogs; D008907:Mink / Q000821:virology*; D053490:Mink enteritis virus / Q000235:genetics / Q000302:isolation & purification*; D010322:Parvoviridae Infections / Q000662:veterinary* / Q000821:virology; D017993:Parvovirus, Canine / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D016415:Sequence Alignment / Q000662:veterinary; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D004279:DNA, Viral
keywords: Canine parvovirus; Mink enteritis virus; PCR-RFLP