Key Laboratory of Special Animal Epidemic Disease, Ministry of Agriculture, PR China, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun 130112, China.
College of Animal Medicine, Agriculture University of Hebei, Baoding 071001, China.
Mol Cell Probes. 2018 Apr;38:7-12. doi: 10.1016/j.mcp.2018.02.004. Epub 2018 Feb 27.
Canine parvovirus (CPV) is an important pathogen in domestic dogs, and the original antigenic types CPV-2 and its variants, CPV-2a, 2b and 2c, are prevalent worldwide. A multiplex TaqMan real-time PCR method was developed for the detection and differentiation of four antigenic types of CPV. A set of primers and probes, CPV-305F/CPV-305R and CPV-2-305P (for CPV-2)/CPV-2a-305P (for CPV-2a, 2b and 2c), was able to differentiate CPV-2 and its variants (CPV-2a, 2b and 2c). Another set of primers and probes, CPV-426F/CPV-426R and CPV-2-426P (for CPV-2 and 2a)/CPV-2b-426P (for CPV-2b)/CPV-2c-426P (for CPV-2c), was able to differentiate CPV-2a (2), CPV-2b, and CPV-2c. With these primers and probes, the multiplex TaqMan real-time PCR assay detected effectively and differentiated CPV-2, 2a, 2b and 2c by two separate real-time PCRs. No cross reactivity was observed with canine distemper virus, canine adenovirus, and canine coronavirus. The detection limit of the assay is 10 genome copies/μL for CPV-2, CPV-2a, CPV-2b, and 10 copies/μL for CPV-2c. The multiplex real-time PCR has 100% agreement with DNA sequencing. We provide a sensitive assay that simultaneously detects and differentiate four antigenic types of CPV and the method was also used for quantification of CPVs viral genome.
犬细小病毒（CPV）是一种重要的犬科病原体，其原始抗原型 CPV-2 及其变体 CPV-2a、2b 和 2c 在全球范围内广泛流行。本研究开发了一种用于检测和区分 CPV 四种抗原型的多重 TaqMan 实时 PCR 方法。一组引物和探针，CPV-305F/CPV-305R 和 CPV-2-305P（用于 CPV-2）/CPV-2a-305P（用于 CPV-2a、2b 和 2c），能够区分 CPV-2 和其变体（CPV-2a、2b 和 2c）。另一组引物和探针，CPV-426F/CPV-426R 和 CPV-2-426P（用于 CPV-2 和 2a）/CPV-2b-426P（用于 CPV-2b）/CPV-2c-426P（用于 CPV-2c），能够区分 CPV-2a（2）、CPV-2b 和 CPV-2c。使用这些引物和探针，多重 TaqMan 实时 PCR 检测方法能够通过两次独立的实时 PCR 有效地检测和区分 CPV-2、2a、2b 和 2c。该检测方法与犬瘟热病毒、犬腺病毒和犬冠状病毒无交叉反应。该检测方法的检测限为 CPV-2、CPV-2a、CPV-2b 为 10 基因组拷贝/μL，CPV-2c 为 10 拷贝/μL。该多重实时 PCR 与 DNA 测序具有 100%的一致性。我们提供了一种敏感的检测方法，可同时检测和区分 CPV 的四种抗原型，该方法还用于 CPV 病毒基因组的定量。
