Abstract
BackgroundCanine distemper virus (CDV) is an enveloped negative-strand RNA virus that exhibits a high mutation rate and continuously expands the range of hosts. Notably, CDV has infected giant panda with spill over from viral reservoirs in canines. Giant pandas (Ailuropoda melanoleuca), especially captive pandas, are known to be susceptible to natural infection with CDV. The high fatality rate of CDV poses a serious threat to the safety of the giant panda population. However, vaccines or drugs for canine distemper in giant pandas have not been developed to date. Therefore, a rapid test that can achieve accurate onsite detection of CDV is important to enable the timely implementation of control measures. In this study, we established a nucleic acid visualization assay for targeting the CDV N gene by using combines reverse transcription recombinase polymerase amplification with a closed vertical flow visualization strip (RT-RPA-VF).ResultsThe RT-RPA-VF assay does not require sophisticated equipment, and it was determined to provide rapid detection at 35 °C for 30 min, while the limit of detection was 5 × 101 copies/μl RNA transcripts and 100.5 TCID50 ml− 1 viruses. The results showed that the assay was high specific to CDV and had no cross-reactivity with other viruses infecting the giant panda. Compared with RT-qPCR, RT-RPA-VF assay had a sensitivity of 100% and a specificity of 100% in 29 clinical samples. The coincidence rate between RT-RPA-VF and RT-qPCR was 100% (kappa = 1), indicating that the RT-RPA-VF assay possessed good diagnostic performance on clinical samples.ConclusionsThe RT-RPA-VF provides a novel alternative for the simple, sensitive, and specific identification of CDV and showed great potential for point of care diagnostics for captive and wild giant panda.
Highlights
Canine distemper virus (CDV) is an enveloped negative-strand Ribonucleic acid (RNA) virus that exhibits a high mutation rate and continuously expands the range of hosts
Canine distemper (CD) is a highly contagious systemic viral disease caused by canine distemper virus (CDV); this disease was initially described as an infectious disease of domestic dogs and has been known since 1760 [1]
Optimizing the RT-Recombinase polymerase amplification (RPA)-VF reaction conditions To improve sensitivity, the RNA transcripts were employed as templates for optimizing the reaction conditions of the RT-RPA-VF assay for CDV, including temperatures and times
Summary
Canine distemper virus (CDV) is an enveloped negative-strand RNA virus that exhibits a high mutation rate and continuously expands the range of hosts. Giant pandas (Ailuropoda melanoleuca), especially captive pandas, are known to be susceptible to natural infection with CDV. The high fatality rate of CDV poses a serious threat to the safety of the giant panda population. Among all factors threatening the current global population of endangered animals, CDV is one of the leading infectious disease killer, posing a serious long-term threat [6, 7]. It was reported as early as 1997 that CDV caused the deaths of captive giant pandas when three pandas died at Chongqing Zoo [6]. We have reported CDV outbreaks in the giant panda with a morbidity of 27% and mortality of 23% in the Shaanxi Rare Wild Animal Rescue and Breeding Research Center, China, between December 2014 and April 2015 [8]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
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.
