Method for the elucidation of LAMP products captured on lateral flow strips in a point of care test for HPV 16
Date
2020-09
Authors
Landaverde, Lena
Wong, Winnie
Hernandez, Gabriela
Fan, Andy
Klapperich, Catherine
Version
Accepted manuscript
OA Version
Citation
L. Landaverde, W. Wong, G. Hernandez, A. Fan, C. Klapperich. 2020. "Method for the elucidation of LAMP products captured on lateral flow strips in a point of care test for HPV 16.." Anal Bioanal Chem, Volume 412, Issue 24, pp. 6199 - 6209. https://doi.org/10.1007/s00216-020-02702-9
Abstract
Loop-mediated amplification (LAMP) is an isothermal amplification technique favored in diagnostics and point-of-care work due to its high sensitivity and ability to run in isothermal conditions. In addition, a visual readout by lateral flow strips (LFS) can be used in conjunction with LAMP, making the assay accessible at the point-of-care. However, the amplicons resulting from a LAMP reaction varied in length and shape, making them undiscernible on a double-stranded DNA intercalating dye stained gel. Standard characterization techniques also do not identify which amplicons specifically bind to the LFS, which generate the visual readout. We aimed to standardize our characterization of LAMP products during assay development by using fluorescein amidite (FAM) and biotin-tagged loop forward and backward primers during assay development. A pvuII restriction enzyme digest is applied to the LAMP products. FAM-tagged bands are directly correlated with the LFS visual readout. We applied this assay development workflow for an HPV 16 assay using both plasmid DNA and clinical samples to demonstrate proof of concept for generalized assay development work.
Description
License
Copyright © Springer-Verlag GmbH Germany, part of Springer Nature 2020. This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic.