Development of a recombinase polymerase amplification lateral flow dipstick (RPA-LFD) for the field diagnosis of caprine arthritis-encephalitis virus (CAEV) infection.
Caprine arthritis-encephalitis (CAE) in goats is a complex disease syndrome caused by a lentivirus. This persistent viral infection results in arthritis in adult goats and encephalitis in lambs. The prognosis for the encephalitic form is normally poor, and this form of the disease has caused substantial economic losses for goat farmers. Hence, a more efficient detection platform based on recombinase polymerase amplification (RPA) and a lateral flow dipstick (LFD) was developed in the present study for detecting the proviral DNA of caprine arthritis-encephalitis virus (CAEV). Under the optimal incubation conditions, specifically, 30min at 37°C for RPA followed by 5min at room temperature for LFD, the assay was found to be sensitive to a lower limit of 80pg of total DNA and 10 copies of plasmid DNA. Furthermore, there was no cross-reaction with other tested viruses, including goat pox virus and bovine leukemia virus. Given its simplicity and portability, this RPA-LFD protocol can serve as an alternative tool to ELISA for the primary screening of CAEV, one that is suitable for both laboratory and field application. When the RPA-LFD was applied in parallel with serological ELISA for the detection of CAEV in field samples, the RPA-LFD assay exhibited a higher sensitivity than the traditional method, and 82% of the 200 samples collected in Taiwan were found to be positive. To our knowledge, this is the first report providing evidence to support the use of an RPA-LFD assay as a specific and sensitive platform for detecting CAEV proviral DNA in goats in a faster manner, one that is also applicable for on-site utilization at farms and that should be useful in both eradication programs and epidemiological studies.
Henchung Branch, Livestock Research Institute, Council of Agriculture, Executive Yuan, No.1, Muchang Rd., Hengchun Township, Pingtung County 946, Taiwan; Hsinchu Branch, Livestock Research Institute, Council of Agriculture, Executive Yuan, No. 207-5, Bi-tou-mian, Wu-hoo village, Si-hoo Township, Miao-li County 36848, Taiwan; Department of Animal Science and Technology, National Taiwan University, No. 50, Ln. 155, Sec. 3, Keelung Rd., Taipei City 10672, Taiwan.
Po-An
PA
Tu
Department of Animal Science and Technology, National Taiwan University, No. 50, Ln. 155, Sec. 3, Keelung Rd., Taipei City 10672, Taiwan; Agriculture Experiment Station, College of Bioresources and Agriculture, National Taiwan University No. 50, Ln. 155, Sec. 3, Keelung Rd., Taipei City 10672, Taiwan.
Jia-Shian
JS
Shiu
National Institute for Animal Health, Council of Agriculture, Executive Yuan, No. 376, Zhongzheng Rd., New Taipei City 251, Taiwan.
Shu-Hwae
SH
Lee
Graduate Institute of Molecular and Comparative Pathobiology, School of Veterinary Medicine, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei 10617, Taiwan; Graduate Institute of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei 10617, Taiwan.
Victor Fei
VF
Pang
Henchung Branch, Livestock Research Institute, Council of Agriculture, Executive Yuan, No.1, Muchang Rd., Hengchun Township, Pingtung County 946, Taiwan.
De-Chi
DC
Wang
Department of Animal Science and Technology, National Taiwan University, No. 50, Ln. 155, Sec. 3, Keelung Rd., Taipei City 10672, Taiwan. Electronic address: demonwang@ntu.edu.tw.
Pei-Hwa
PH
Wang
D000818:Animals; D016088:Arthritis-Encephalitis Virus, Caprine / Q000235:genetics / Q000302:isolation & purification*; D002845:Chromatography / Q000379:methods*; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D015511:Goat Diseases / Q000175:diagnosis* / Q000821:virology*; D006041:Goats; D016180:Lentivirus Infections / Q000175:diagnosis / Q000662:veterinary*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D019095:Point-of-Care Systems; D011533:Proviruses / Q000235:genetics; D045522:Recombinases / Q000378:metabolism; D012680:Sensitivity and Specificity; D013624:Taiwan; D013696:Temperature
D004279:DNA, Viral; D045522:Recombinases
Caprine arthritis-encephalitis virus (CAEV); Lateral flow dipstick (LFD); Recombinase polymerase amplification (RPA)