STORRE
STORRE: Stirling Online Research Repository
http://hdl.handle.net/1893/31109| Appears in Collections: | Aquaculture Book Chapters and Sections |
| Title: | Development and Validation of Real-Time RT-LAMP Assays for the Specific Detection of Zika Virus |
| Author(s): | Lopez-Jimena, Benjamin Bakheit, Mohammed Bekaert, Michaël Harold, Graham Frischmann, Sieghard Fall, Cheikh Diagne, Cheikh Tidiane Faye, Oumar Faye, Ousmane Sall, Amadou Alpha Weidmann, Manfred |
| Contact Email: | m.w.weidmann@stir.ac.uk |
| Editor(s): | Kobinger, Gary Racine, Trina |
| Sponsor: | European Commission (Horizon 2020) |
| Citation: | Lopez-Jimena B, Bakheit M, Bekaert M, Harold G, Frischmann S, Fall C, Diagne CT, Faye O, Faye O, Sall AA & Weidmann M (2020) Development and Validation of Real-Time RT-LAMP Assays for the Specific Detection of Zika Virus. In: Kobinger G & Racine T (eds.) Zika Virus: Methods and Protocols. Methods in Molecular Biology, 2142. New York: Springer, pp. 147-164. https://doi.org/10.1007/978-1-0716-0581-3_13 |
| Keywords: | Zika virus Isothermal amplification Real-time detection RT-LAMP LAVA software Primer Explorer V4 |
| Issue Date: | 2020 |
| Date Deposited: | 7-May-2020 |
| Series/Report no.: | Methods in Molecular Biology, 2142 |
| Abstract: | Two one-step real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays for the detection of Zika virus (ZIKV) were developed, based on two different primer design approaches: (1) open source, based on a combination of sequence diversity clustering (phylogeny and principal component analysis) and LAVA algorithm, using 45 whole genome ZIKV sequences retrieved from the National Center for Biotechnology Information (NCBI) database; (2) standard software for LAMP primer design (Primer Explorer V4), using 59 sequences of the ZIKV 3′ UTR. The assays were firstly evaluated with External Quality Assessment panels from INSTAND e.V. (Germany) and EVD-LabNet (The Netherlands) including 4 and 12 unknown samples, respectively, and secondly, with 9 human, mosquito, and monkey ZIKV isolates from Africa (Senegal, Ivory Coast, and Uganda) and America (Brazil). The limit of detection as determined by probit analysis was 181 molecules for both RT-LAMP assays, and 100% reproducibility in the assays was obtained for 103 molecules (4/8 repetitions were positive for 102 molecules). Both assays were specific, amplifying only ZIKV RNA and not cross-detecting other arboviruses included in this study. |
| Rights: | The publisher does not allow this work to be made publicly available in this Repository. Please use the Request a Copy feature at the foot of the Repository record to request a copy directly from the author. You can only request a copy if you wish to use this work for your own research or private study. |
| DOI Link: | 10.1007/978-1-0716-0581-3_13 |
| Licence URL(s): | http://www.rioxx.net/licenses/under-embargo-all-rights-reserved |
| File | Description | Size | Format | |
|---|---|---|---|---|
| Lopez-Jimena et al 2020_CHAPTER_13.pdf | Fulltext - Published Version | 490.19 kB | Adobe PDF | Under Permanent Embargo Request a copy |
Note: If any of the files in this item are currently embargoed, you can request a copy directly from the author by clicking the padlock icon above. However, this facility is dependent on the depositor still being contactable at their original email address.
This item is protected by original copyright |
Items in the Repository are protected by copyright, with all rights reserved, unless otherwise indicated.
The metadata of the records in the Repository are available under the CC0 public domain dedication: No Rights Reserved https://creativecommons.org/publicdomain/zero/1.0/
If you believe that any material held in STORRE infringes copyright, please contact library@stir.ac.uk providing details and we will remove the Work from public display in STORRE and investigate your claim.