Yujun Zhu

Zhejiang University

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Miaoli Wu

Guangdong Laboratory Animals Monitoring Institute

Yuexiao Lian

Guangdong Laboratory Animals Monitoring Institute

Bihong Huang

International Monetary Fund

Xinchao Liu

Peking Union Medical College Hospital

Xiao Cong

Guangdong Laboratory Animals Monitoring Institute

Yue Shi

Chinese PLA General Hospital

Yinzhu Luo

Guangdong Laboratory Animals Monitoring Institute

Youfang Gu

Anhui Science and Technology University

Feng Cong

Guangdong Laboratory Animals Monitoring Institute

Mengdi Zhang

Zhejiang Chinese Medical University

Cooperative Institutions

Guangdong Laboratory Animals Monitoring Institute

Zhejiang University

Wenzhou Vocational College of Science and Technology

Anhui Science and Technology University

Anhui University of Science and Technology

Chinese Academy of Agricultural Sciences

Harbin Veterinary Research Institute

South China Agricultural University

Chinese Academy of Medical Sciences & Peking Union Medical College

Tongji University

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Establishment of a recombinase polymerase amplification (RPA) fluorescence assay for the detection of swine acute diarrhea syndrome coronavirus (SADS-CoV)

Research Square (Research Square) (2022)

Xiao Cong Yujun Zhu Xinchao Liu Yuexiao Lian Bihong Huang

Abstract Background: Swine acute diarrhea syndrome coronavirus (SADS-CoV) causes acute vomiting and diarrhea of piglets, leading to significant financial losses in pig industry. Recombinase polymerase amplification (RPA) technology is the second method for nucleic acid amplification under constant temperature conditions besides loop-mediated isothermal amplification (LAMP). The study established a real-time reverse transcription (RT)-RPA assay for early confirmatory diagnoses to detection SADS-CoV. Results: The detection limit of the real-time RT-RPA was 74 copies/µL of SADS-CoV genomic RNA standard in 95% of cases. The assay was performed in less than 30 min and no cross-reactions were observed with 8 other common viruses that affect swine, namely, CSFV, PRRSV, PRV, SIV, SVA, TGEV, PEDV, or PDCoV. The coefficient of variation (C.V.) values of the two standards dilutions and a positive clinical sample ranged from 0 to 4.5%. A total of 72 clinical fecal samples from swine with diarrheal symptoms were analyzed via the developed RT-RPA and qRT-PCR. There was 98.61% agreement between the RT-RPA and the qRT-PCR results. Conclusions: These results indicate that the developed RT-RPA assay has good specificity, sensitivity, stability, and repeatability. In summary, the established RT-RPA assay could satisfy the demand for infield diagnoses, and is suitable for use in remote areas as it is fast, portable, and cost-effective.

Recombinase Polymerase Amplification

Coronavirus

Serial dilution

10.21203/rs.3.rs-1385813/v1

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The Establishment of a Recombinant Polymerase Amplification Technique for the Detection of Mouse Poxvirus

Research Square (Research Square) (2023)

Yuexiao Lian Mengdi Zhang Yujun Zhu Miaoli Wu Bihong Huang

Abstract Background: Ectromelia virus (ECTV) is the pathogen of mousepox, which has a limited host range and a high mortality rate. In the past century, ECTV has affected laboratory mouse colonies worldwide. Recombinant polymerase amplification (RPA), which is widely used in virus detection, is an isothermal amplification method. Results: In this study, a probe-based RPA detection method was established for rapid and sensitive detection of ECTV.Primers were designed for the highly conserved region of crmD gene, the main core protein of recessive poxvirus, and standard plasmids were constructed.The sensitivity of the RPA assay is 100 copies of the genome per reaction. In addition, the method showed high specificity and did not cross-react with other common mouse viruses.Therefore, the practicability of the RPA method in the field was confirmed by the detection of 52 clinical samples. The real-time RPA assay was very similar to the ECTV real-time PCR assay, with 100% agreement. Conclusions: In conclusion, this RPA assay, especially in low-resource settings, provides a novel alternative for sensitive, simple, and specific identification of ECTV.

Ectromelia virus

10.21203/rs.3.rs-2343023/v1

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Additional file 3 of Establishment of a recombinase polymerase amplification (RPA) fluorescence assay for the detection of swine acute diarrhea syndrome coronavirus (SADS-CoV)

Figshare (2022)

Xiao Cong Yujun Zhu Xinchao Liu Yuexiao Lian Bihong Huang

Additional file 3: Supplementary Table 1. The detection results of 11 samples by quantitative real-time PCR assays and the RT-PCR system.

Recombinase Polymerase Amplification

Coronavirus

10.6084/m9.figshare.21316179

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