A Rapid Real Time Fluorescence Recombinase-aided Amplification Method for Detecting Porcine Pseudorabies Virus

XIE Mingjie; KANG Longbin; CHEN Qiuyong; WU Xuemin; WANG Longbai; ZHOU Lunjiang; LIU Yutao

doi:10.19303/j.issn.1008-0384.2024.07.001

Volume 39 Issue 7

Jul. 2024

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Article Navigation > Fujian Journal of Agricultural Sciences > 2024 > 39(7): 753-758

XIE M J, KANG L B, CHEN Q Y, et al. A Rapid Real Time Fluorescence Recombinase-aided Amplification Method for Detecting Porcine Pseudorabies Virus [J]. Fujian Journal of Agricultural Sciences，2024，39（7）：753−758 doi: 10.19303/j.issn.1008-0384.2024.07.001

Citation:

XIE M J, KANG L B, CHEN Q Y, et al. A Rapid Real Time Fluorescence Recombinase-aided Amplification Method for Detecting Porcine Pseudorabies Virus [J]. Fujian Journal of Agricultural Sciences，2024，39（7）：753−758 doi: 10.19303/j.issn.1008-0384.2024.07.001

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A Rapid Real Time Fluorescence Recombinase-aided Amplification Method for Detecting Porcine Pseudorabies Virus

doi: 10.19303/j.issn.1008-0384.2024.07.001

XIE Mingjie^{1, 2
,},
KANG Longbin^{1, 2},
CHEN Qiuyong²,
WU Xuemin²,
WANG Longbai²,
ZHOU Lunjiang^{2
,
,},
LIU Yutao^{2
,
,}

1.
School of Animal Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China
2.
Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences/Fujian Provincial Engineering and Technology Research Center for Livestock and Poultry Disease Prevention and Control, Fuzhou, Fujian　350013, China

Received Date: 2024-03-11
Rev Recd Date: 2024-06-14

Available Online: 2024-08-15

Publish Date: 2024-07-01

Abstract

Abstract

Objective A rapid method applying the recombinase-aided amplification (RAA) method for detecting porcine pseudorabies virus (PRV) was developed. Methods Based on the sequence of gE gene in PRV, specific primers and probes were designed. Amplification conditions were optimized, and assay specificity, sensitivity, and reproducibility scrutinized by a verification trial on clinical samples. Results The newly developed assay successfully amplified the PRV nucleic acids in merely 23m under the constant temperature of 43 ℃ with a detection limit of 111 copies·μL⁻¹. There were no cross reactions with viruses that produced reproductive and respiratory syndromes, epidemic diarrhea, rotavirus, transmissible gastroenteritis, circovirus 2, or circovirus 3 on pigs. The coefficients of variation within a group and between groups on the repeatability test were less than 5%. And, on 40 clinical samples, the positive detection on PRV of the assay was 15% (6/40), which was comparable to that of the conventional PCR. Conclusion A simple, rapid, efficient, and accurate method of fluorescence RAA detection on PRV was established for laboratory testing and epidemiological investigation of the disease.
- Porcine pseudorabies virus,
- recombinase-aided amplification,
- detection method

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References(20)

References

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