A <i>Taq</i>Man RT-qPCR Assay for Gyrovirus 3 Detection

ZHUANG Liyun; CHEN Weichen; DAI Tingting; LEI Tianyu; CHEN Haiyu; ZHENG Xintian

doi:10.19303/j.issn.1008-0384.2023.09.003

Volume 38 Issue 9

Sep. 2023

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Article Navigation > Fujian Journal of Agricultural Sciences > 2023 > 38(9): 1024-1029

ZHUANG L Y, CHEN W C, DAI T T, et al. A TaqMan RT-qPCR Assay for Gyrovirus 3 Detection [J]. Fujian Journal of Agricultural Sciences，2023，38（9）：1024−1029 doi: 10.19303/j.issn.1008-0384.2023.09.003

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ZHUANG L Y, CHEN W C, DAI T T, et al. A TaqMan RT-qPCR Assay for Gyrovirus 3 Detection [J]. Fujian Journal of Agricultural Sciences，2023，38（9）：1024−1029 doi: 10.19303/j.issn.1008-0384.2023.09.003

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A TaqMan RT-qPCR Assay for Gyrovirus 3 Detection

doi: 10.19303/j.issn.1008-0384.2023.09.003

ZHUANG Liyun^{1, 2
,},
CHEN Weichen¹,
DAI Tingting^{1, 2},
LEI Tianyu^{1, 2},
CHEN Haiyu^{1, 2},
ZHENG Xintian^{1, 3
,
,}

1.
Institute of Life Science, Longyan University, Longyan, Fujian　364000, China
2.
College of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou, Fujian　350000, China
3.
Key Laboratory of Fujian Universities Preventive Veterinary Medicine and Biotechnology, Longyan University, Longyan, Fujian　364000, China

Received Date: 2023-06-14
Rev Recd Date: 2023-08-31

Available Online: 2023-10-25

Publish Date: 2023-09-28

Abstract

Abstract

Objective An RT-qPCR method to accurately detect gyrovirus 3 (GyV3) was established. Methods A pair of specific primers and an FAM-labeled fluorophobe were designed according to the conserved region of GyV3 VP2 in Genbank database. Reaction conditions of a rapid TaqMan RT-qPCR assay were optimized, and specificity, sensitivity, repeatability, and simulation tests conducted to verify the applicability of the methodology. Results The assay exhibited a high sensitivity with a minimum detection limit of 1.585 copies·μL⁻¹, a specificity free of cross-reactivity with other prevalent avian pathogens, and a reproducibility with less than 1% coefficients of variation on intra- and inter-batch determinations. On a simulated sample with an added recombinant plasmid to the chicken liver tissue DNA, the assay positively identified GyV3 with same result as did the SYBR Green Ⅰ PCR. Conclusion The newly developed TaqMan RT-qPCR assay showed high sensitivity, specificity, repeatability, and rapid turn-around time in detecting GyV3. It was considered appropriate for clinical diagnosis and epidemiological investigation on the virus.
- Gyrovirus 3,
- RT-qPCR,
- detection assay,
- transmissible viral proventriculitis

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References(25)

References

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