Establishment of the real-time RAA detection method for Gyrovirus galga 1
In order to establish a rapid detection method for Gyrovirus galga 1(GyVg1),the specific primers and probe were designed based on the conserved region of GyVg1 by searching and comparing se-quences currently available in GenBank.After screening primers and optimization of the reaction con-ditions,a recombinase acid amplication(RAA)detection method for GyVg1 was established.The results showed that GyVg1 could be identified in 20 minutes by the real-time fluorescence RAA assay.This method could only distinguish GyVg1,but not react with Gyrovirus homsa1(GyH1),chicken infectious anemia virus(CIAV),fowl adenovirus 4(FAdV4),H9 subtype avian influenza virus(H9-AIV),Newcastle disease virus(NDV),avian infectious bronchitis virus(IBV),infectious bursal disease virus(IBDV),chicken parvovirus(ChPV),avian leukosis virus(ALV),Mycoplasma gallisepticum(MG)and Mycoplasma synoviae(MS);The lowest limit of detection was 1 × 102 copies/μL and the coefficient of variations in intra-and inter-assay were both less than 6％.231 samples were tested simultaneously by fluorescence RAA and fluorescence quantitative PCR,and the results were consistent with each other.In conclusion,a re-al-time fluorescence RAA method is successfully established for the first time and this method has good specificity,sensitivity and repeatability,which provides a new technical means for the accurate and rapid detection of GyVg1 infection.
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