Abstract
With increasing complications of canine parvovirus infection cases, disease diagnosis and treatment have become more difficult. In this study, specificity primers for the conserved region of the VP2 gene of canine parvovirus 2 (CPV-2) were synthesized and evaluated. An improved polymerase cross-linking spiral reaction (PCLSR) method for early and rapid diagnosis of CPV-2 was established. The results showed that the amplification reaction was optimal when run at 62°C for 50 min and could be used to detect CPV-2 without any cross-reactions with other pathogens of canine infectious diseases. Reaction results were directly judged by the naked eyes, with the positive amplification tube shown as luminous yellow and the negative tube as bright purple. Compared with the previously reported polymerase spiral reaction (PSR) method for CPV-2 detection, this reaction was performed using improved primer pairs and a better dye identification method (using an indicator comprising phenol red and cresol red). The detection limit of PCLSR was 3.9 × 101 copies using gel electrophoresis or a visible dye. The positive rate of 132 clinical samples was 42.42%, which was identically the same as that of the PSR method and slightly higher than that of the colloidal gold strip method (39.39%). The newly developed CPV-PCLSR assay shows the advantage of rapid visualization of results and offers a convenient and rapid method for early CPV-2 diagnosis with higher sensitivity and specificity than the established methods.
Highlights
Canine parvovirus 2 (CPV-2) belongs to the species Carnivore protoparvovirus 1, genus Parvovirus of the family Parvoviridae
All canine infectious disease samples are stored at the Henan Provincial Engineering Laboratory of Insects Bio-reactor, Nanyang Normal University, China, and included canine parvovirus 2 (CPV-2); three CPV-2 variants (CPV-2a, MK518002, CPV-2b, MK517983, and CPV-2c, MK517966); and canine distemper virus (CDV), canine coronavirus (CCoV), canine circovirus (CCV), canine adenovirus-1 (CAV-1), and canine adenovirus-2 (CAV-2)-positive samples; and 59 CPV-2 isolates were determined by virus isolation and VP2 sequencing that were collected from 2016 to 2019 (CPV-2a, n = 17; CPV-2b, n = 12; and CPV-2c, n = 30)
Assay data used for optimum reaction temperature and time for CPV-2 diagnosis by polymerase cross-linking spiral reaction (PCLSR) are displayed in detail in Supplementary Table 1
Summary
Canine parvovirus 2 (CPV-2) belongs to the species Carnivore protoparvovirus 1, genus Parvovirus of the family Parvoviridae. It is a single-stranded linear DNA virus, which was first isolated from infected dogs in the United States in 1978 and subsequently spread worldwide [1]. In China, CPV-2 was first isolated and reported in 1983, and infections have been regularly reported ever since. The morbidity and mortality rates of CPV-2 infection are as high as 70% [2]. CPV-2 is one of the most threatening pathogens for domestic and wild dogs.
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
