Abstract
Feline leukemia virus (FeLV) is a major viral disease in cats, causing leukemia and lymphoma. The molecular detection of FeLV RNA and the DNA provirus are important for staging of the disease. However, the rapid immunochromatographic assay commonly used for antigen detection can only detect viremia at the progressive stage. In this study, nested recombinase polymerase amplification (nRPA) was developed for exogenous FeLV DNA provirus detection, and reverse transcriptase polymerase amplification (RT-RPA) was developed for the detection of FeLV RNA. The approaches were validated using 108 cats with clinicopathologic abnormalities due to FeLV infection, and from 14 healthy cats in a vaccination plan. The nRPA and RT-RPA assays could rapidly amplify the FeLV template, and produced high sensitivity and specificity. The FeLV detection rate in regression cats by nRPA was increased up to 45.8% compared to the rapid immunochromatographic assay. Hence, FeLV diagnosis using nRPA and RT-RPA are rapid and easily established in low resource settings, benefiting FeLV prognosis, prevention, and control of both horizontal and vertical transmission.
Highlights
Feline leukemia virus (FeLV) is an enveloped positive single-stranded retrovirus which is widely distributed in domestic cats, and occurs occasionally in closely related wild felids[1]
Progressive infections are categorized by detectable antigenemia, FeLV RNA, and DNA provirus, since the virus persistently sheds into the circulation and tissues
The pGEM-T vector harboring the 145 bp of the U3 long terminal repeat (U3LTR) FeLV fragment was used as a template to optimize the nested recombinase polymerase amplification (nRPA) assay
Summary
Feline leukemia virus (FeLV) is an enveloped positive single-stranded retrovirus which is widely distributed in domestic cats, and occurs occasionally in closely related wild felids[1]. Progressive infections are categorized by detectable antigenemia, FeLV RNA, and DNA provirus, since the virus persistently sheds into the circulation and tissues. Such progressive stage infected cats are a major reservoir for horizontal transmission. In this paper we described the development and validation of nRPA and reverse transcriptase (RT-RPA) assays for the detection of FeLV DNA provirus and RNA. Both FeLV RPA assays can be used in veterinary hospitals for rapid diagnosis, which is required in order to enable effective prevention and infection control
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