RPB0547

Pathogen Description

Target Pathogen Pathogen Name NCBI Taxonomy ID Order Family Genus Species Pathogen type
Tick-Borne Encephalitis Virus Tick-borne encephalitis virus,TBEV,FSME virus,Tick born encephalitis virus,tick-borne encephalitis virus TBE virus,tick-borne encephalitis virus TBEV 11084 Amarillovirales Flaviviridae Orthoflavivirus Orthoflavivirus encephalitidis Viruses

Primer Description

Primer Name Sequence(5'-3') Length(bp) Primer Final Concentration(μM） GC Content（%） Predicted Melting Temperature(℃） Molecular Weight(g/moles) Positions in GenBank accession number
RAA-F CCTTTGGACAGCAGCGAGTGTTCAARGAGAA 31 \ 50 64.87 9601.3 \
RAA-R CTATGAAYTCCTCTCTGCTGCACATTCGTGG 31 \ 50 63.32 9420.66 \
RAA-P AGGCTCAGGAGCCTCAGCCTGGCACAARGG[FAM-dT](THF)A [BHQ-dT] CATGAGAGCAGTGAATGA (C3-spacer) 49 \ 56.12 75.42 15230.93 \

Gene Description

Target Gene GenBank ID
NS5 gene \

Assay Description

Application Assay Primer Designing Software Reaction Time(min) Assay Temperature(℃) Readout System(s) Limit of Detection(LoD) Sensitivity(%) Specificity(%)
The RT-RAA assay developed is as good as an RT-qPCR assay, indicating that it is a valuable addition to the serological tests currently available for the diagnosis of TBE. This technique should attract more attention and undergo further development in the near future. RT-RAA Primer Express 3.0 30 min 39 °C \ 20 copies per reaction \ 1

Publication Description

Year of Publication Title Author(s) Journal PMID DOI
2019 A Reverse-transcription Recombinase-aided Amplification Assay for the Rapid Detection of the Far-Eastern Subtype of Tick-borne Encephalitis Virus Qian Ying Wang,Fan Li,Xin Xin Shen,Shi Hong Fu,Ying He,Wen Wen Lei,Guo Dong Liang,Huan Yun Wang,Xue Jun Ma Biomedical and environmental sciences : BES 31217052 10.3967/bes2019.047

A Reverse-transcription Recombinase-aided Amplification Assay for the Rapid Detection of the Far-Eastern Subtype of Tick-borne Encephalitis Virus

Author(s):

Qian Ying Wang,Fan Li,Xin Xin Shen,Shi Hong Fu,Ying He,Wen Wen Lei,Guo Dong Liang,Huan Yun Wang,Xue Jun Ma

Journal:

Biomedical and environmental sciences : BES

Year:

2019

Abstract:

Objective: Tick-borne encephalitis virus (TBEV) is an emerging pathogen in Europe and North Asia that causes tick-borne encephalitis (TBE). A simple, rapid method for detecting TBEV RNA is needed to control this disease. Methods: A reverse-transcription recombinase-aided amplification (RT-RAA) assay was developed. This assay can be completed in one closed tube at 39 °C within 30 minutes. The sensitivity and specificity of RT-RAA were validated using non-infectious synthetic RNA representing a fragment of the NS5 region of the wild-type (WT) TBEV genome and the Senzhang strain. Additionally, 10 batches of tick samples were used to evaluate the performance of the RT-RAA assay. Results: The analytical limit of detection of the assay was 20 copies per reaction of the TBEV synthetic transcript and 3 plaque-forming units (pfu) per reaction of TBEV titers. With the specific assay, no signal due to other arboviruses was observed. Of the 10 batches of tick samples obtained from the Changbai Mountains of China, three were TBEV-positive, which was consistent with the results of the quantitative real-time PCR assay. Conclusion: A rapid, highly sensitive, specific, and easy-to-use method was developed for the detection of the TBEV Far-Eastern subtype.