Rapid detection of Banna virus by reverse transcription-loop-mediated isothermal amplification (RT-LAMP)

Han Xia; Na Zhao; Lu Zhao; Yujuan Wang; Wang Zhao; Zhiming Yuan

doi:10.1016/j.ijid.2018.10.025

Rapid detection of Banna virus by reverse transcription-loop-mediated isothermal amplification (RT-LAMP)

Int J Infect Dis. 2019 Jan:78:93-98. doi: 10.1016/j.ijid.2018.10.025. Epub 2018 Nov 3.

Authors

Han Xia¹, Na Zhao², Lu Zhao², Yujuan Wang³, Wang Zhao⁴, Zhiming Yuan⁵

Affiliations

¹ Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, 430071, China. Electronic address: hanxia@wh.iov.cn.
² Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, 430071, China; University of Chinese Academy of Sciences, Beijing, 100049, China.
³ Henan Province OriginBio Biotechnology Co., LtD., Zhengzhou, Henan, 450000, China.
⁴ Shanghai Cohere Electronic Technology Co., LtD., Shanghai, 201101, China.
⁵ Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, Hubei, 430071, China. Electronic address: yzm@wh.iov.cn.

PMID: 30395978
DOI: 10.1016/j.ijid.2018.10.025

Abstract

Objectives: Banna virus (BAV) is classified in the genus Seadornavirus within the Reoviridae family and considered to be an emerging pathogen. We aimed to develop a rapid and simple molecular detection approach for all BAV subgroups in isothermal conditions.

Method: A set of six specific primers was designed to target the segment 12 of BAV, and the reverse transcription-loop mediated isothermal amplification (RT-LAMP) assay was developed and compared with conventional RT-PCR method.

Results: The amplification of the RT-LAMP assay can be obtained within 40min at 65°C. The results from specificity showed that only target BAVs RNA including genotypes A, B and C were amplified and the assay demonstrated a sensitivity of 3.6×10^-2PFU/mL, which was higher than conventional RT-PCR measurement. A good reliability for the assay was presented in the further evaluation for BAVs RNA from serial diluted BAV-spiked serum and 47 pools of field mosquito samples.

Conclusions: Our findings present a rapid, sensitive and specific RT-LAMP assay that can be applied for BAV detection in clinical or field samples in the future.

Keywords: Banna virus; Detection; Mosquito; RT-LAMP.

MeSH terms

Aedes
Animals
Coltivirus / isolation & purification*
Culex
Humans
Nucleic Acid Amplification Techniques / methods*
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction
Reverse Transcription