Early detection of dengue virus by use of reverse transcription-recombinase polymerase amplification
A method for the rapid diagnosis of early dengue virus (DENV) infection is highly needed. Here, a prototype reverse transcription-recombinase polymerase amplification (RT-RPA) assay was developed. The assay detected DENV RNA in <20 min without the need for thermocycling amplification. The assay e...
| Main Authors: | , , , , , , , , , , , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2015
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| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/101099 http://hdl.handle.net/10220/25718 |
| _version_ | 1811678313040576512 |
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| author | Teoh, Boon-Teong Sam, Sing-Sin Tan, Kim-Kee Danlami, Mohammed Bashar Shu, Meng-Hooi Johari, Jefree Hooi, Poh-Sim Brooks, David Piepenburg, Olaf Nentwich, Oliver Wilder-Smith, Annelies Franco, Leticia Tenorio, Antonio AbuBakar, Sazaly |
| author2 | Tang, Y.-W. |
| author_facet | Tang, Y.-W. Teoh, Boon-Teong Sam, Sing-Sin Tan, Kim-Kee Danlami, Mohammed Bashar Shu, Meng-Hooi Johari, Jefree Hooi, Poh-Sim Brooks, David Piepenburg, Olaf Nentwich, Oliver Wilder-Smith, Annelies Franco, Leticia Tenorio, Antonio AbuBakar, Sazaly |
| author_sort | Teoh, Boon-Teong |
| collection | NTU |
| description | A method for the rapid diagnosis of early dengue virus (DENV) infection is highly needed. Here, a prototype reverse transcription-recombinase polymerase amplification (RT-RPA) assay was developed. The assay detected DENV RNA in <20 min without the need for thermocycling amplification. The assay enabled the detection of as few as 10 copies of DENV RNA. The designed RT-RPA primers and exo probe detected the DENV genome of at least 12 genotypes of DENV circulating globally without cross-reacting with other arboviruses. We assessed the diagnostic performance of the RT-RPA assay for the detection of DENV RNA in 203 serum samples of patients with clinically suspected dengue. The sera were simultaneously tested for DENV using a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay, quantitative RT-PCR (qRT-PCR), and IgM- and IgG-capture enzyme-linked immunosorbent assays (ELISA). Acute DENV infection was confirmed in 130 samples and 61 of the samples (46.9%) were classified as viremic with qRT-PCR. The RT-RPA assay showed good concordance (κ of ≥0.723) with the RT-LAMP and qRT-PCR assays in detecting the dengue viremic samples. When used in combination with ELISA, both the RT-RPA and RT-LAMP assays increased the detection of acute DENV infection to ≥95.7% (≥45/47) in samples obtained within 5 days of illness. The results from the study suggest that the RT-RPA assay is the most rapid molecular diagnostic tool available for the detection of DENV. Hence, it is possible to use the RT-RPA assay in a laboratory to complement routine serology testing for dengue. |
| first_indexed | 2024-10-01T02:51:16Z |
| format | Journal Article |
| id | ntu-10356/101099 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T02:51:16Z |
| publishDate | 2015 |
| record_format | dspace |
| spelling | ntu-10356/1010992022-02-16T16:28:58Z Early detection of dengue virus by use of reverse transcription-recombinase polymerase amplification Teoh, Boon-Teong Sam, Sing-Sin Tan, Kim-Kee Danlami, Mohammed Bashar Shu, Meng-Hooi Johari, Jefree Hooi, Poh-Sim Brooks, David Piepenburg, Olaf Nentwich, Oliver Wilder-Smith, Annelies Franco, Leticia Tenorio, Antonio AbuBakar, Sazaly Tang, Y.-W. Lee Kong Chian School of Medicine (LKCMedicine) DRNTU::Science::Biological sciences::Microbiology A method for the rapid diagnosis of early dengue virus (DENV) infection is highly needed. Here, a prototype reverse transcription-recombinase polymerase amplification (RT-RPA) assay was developed. The assay detected DENV RNA in <20 min without the need for thermocycling amplification. The assay enabled the detection of as few as 10 copies of DENV RNA. The designed RT-RPA primers and exo probe detected the DENV genome of at least 12 genotypes of DENV circulating globally without cross-reacting with other arboviruses. We assessed the diagnostic performance of the RT-RPA assay for the detection of DENV RNA in 203 serum samples of patients with clinically suspected dengue. The sera were simultaneously tested for DENV using a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay, quantitative RT-PCR (qRT-PCR), and IgM- and IgG-capture enzyme-linked immunosorbent assays (ELISA). Acute DENV infection was confirmed in 130 samples and 61 of the samples (46.9%) were classified as viremic with qRT-PCR. The RT-RPA assay showed good concordance (κ of ≥0.723) with the RT-LAMP and qRT-PCR assays in detecting the dengue viremic samples. When used in combination with ELISA, both the RT-RPA and RT-LAMP assays increased the detection of acute DENV infection to ≥95.7% (≥45/47) in samples obtained within 5 days of illness. The results from the study suggest that the RT-RPA assay is the most rapid molecular diagnostic tool available for the detection of DENV. Hence, it is possible to use the RT-RPA assay in a laboratory to complement routine serology testing for dengue. Published version 2015-06-02T02:27:20Z 2019-12-06T20:33:22Z 2015-06-02T02:27:20Z 2019-12-06T20:33:22Z 2015 2015 Journal Article Teoh, B.-T., Sam, S.-S., Tan, K.-K., Danlami, M. B., Shu, M.-H., Johari, J., et al. (2015). Early detection of dengue virus by use of reverse transcription-recombinase polymerase amplification. Journal of clinical microbiology, 53(3), 830-837. https://hdl.handle.net/10356/101099 http://hdl.handle.net/10220/25718 10.1128/JCM.02648-14 25568438 en Journal of clinical microbiology © 2015 American Society for Microbiology. This paper was published in Journal of Clinical Microbiology and is made available as an electronic reprint (preprint) with permission of American Society for Microbiology. The paper can be found at the following official DOI: [http://dx.doi.org/10.1128/JCM.02648-14]. One print or electronic copy may be made for personal use only. Systematic or multiple reproduction, distribution to multiple locations via electronic or other means, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper is prohibited and is subject to penalties under law. 8 p. application/pdf |
| spellingShingle | DRNTU::Science::Biological sciences::Microbiology Teoh, Boon-Teong Sam, Sing-Sin Tan, Kim-Kee Danlami, Mohammed Bashar Shu, Meng-Hooi Johari, Jefree Hooi, Poh-Sim Brooks, David Piepenburg, Olaf Nentwich, Oliver Wilder-Smith, Annelies Franco, Leticia Tenorio, Antonio AbuBakar, Sazaly Early detection of dengue virus by use of reverse transcription-recombinase polymerase amplification |
| title | Early detection of dengue virus by use of reverse transcription-recombinase polymerase amplification |
| title_full | Early detection of dengue virus by use of reverse transcription-recombinase polymerase amplification |
| title_fullStr | Early detection of dengue virus by use of reverse transcription-recombinase polymerase amplification |
| title_full_unstemmed | Early detection of dengue virus by use of reverse transcription-recombinase polymerase amplification |
| title_short | Early detection of dengue virus by use of reverse transcription-recombinase polymerase amplification |
| title_sort | early detection of dengue virus by use of reverse transcription recombinase polymerase amplification |
| topic | DRNTU::Science::Biological sciences::Microbiology |
| url | https://hdl.handle.net/10356/101099 http://hdl.handle.net/10220/25718 |
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