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Point‐of‐care diagnostic assay for rapid detection of porcine deltacoronavirus using the recombinase polymerase amplification method
Author(s) -
Ma Lei,
Zeng Fanwen,
Huang Bihong,
Zhu Yujun,
Wu Miaoli,
Xu Fengjiao,
Xiao Li,
Huang Ren,
Ma Jingyun,
Cong Feng,
Guo Pengju
Publication year - 2019
Publication title -
transboundary and emerging diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.392
H-Index - 63
eISSN - 1865-1682
pISSN - 1865-1674
DOI - 10.1111/tbed.13155
Subject(s) - recombinase polymerase amplification , recombinase , point of care testing , point of care , polymerase chain reaction , computational biology , biology , virology , genetics , medicine , gene , pathology , immunology , recombination
Summary Porcine deltacoronavirus ( PDC oV) has emerged and spread throughout the porcine industry in many countries over the last 6 years. PDC oV caused watery diarrhoea, vomiting and dehydration in newborn piglets. A sensitive diagnostic method would be beneficial to the prevention and control of PDC oV infection. Recombinase polymerase amplification ( RPA ) is an isothermal amplification method which has been widely used for virus detection. A probe‐based reverse transcription RPA ( RT ‐ RPA ) assay was developed for real‐time detection of PDC oV. The amplification can be finished in 20 min and fluorescence monitoring was performed by a portable device. The lowest detection limit of the PDC oV RT ‐ RPA assay was 100 copies of RNA molecules per reaction; moreover, the RT ‐ RPA assay had no cross‐reaction with other common swine viruses. The clinical performance of the RT ‐ RPA assay was evaluated using 108 clinical samples (54 intestine specimens and 54 faecal swab specimens). The coincidence rate of the detection results for clinical samples between RT ‐ RPA and RT ‐ qPCR was 97.2%. In summary, the real‐time RT ‐ RPA assay offers a promising alternative to RT ‐ qPCR for point‐of‐care detection of PDC oV.