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Application of a Recombinase Polymerase Amplification (RPA) assay and pilot field testing for Giardia duodenalis at Lake Albert, Uganda.

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Molina-Gonzalez, Sandra J., Bhattacharyya, Tapan, AlShehri, Hajri, Poulton, Kate, Allen, Stephen ORCID: https://orcid.org/0000-0001-6675-249X, Miles, Michael A., Arianitwe, Moses, Tukahebwa, Edridah M., Webster, Bonnie, Stothard, Russell ORCID: https://orcid.org/0000-0002-9370-3420 and Bustinduy, Amaya L. (2020) 'Application of a Recombinase Polymerase Amplification (RPA) assay and pilot field testing for Giardia duodenalis at Lake Albert, Uganda.'. Parasites & Vectors, Vol 13, Issue 289.

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Abstract

Giardia duodenalis is a common gastrointestinal protozoan causing 184 million cases of giardiasis worldwide annually. Detection is by microscopy or coproantigen assays, although diagnostic sensitivity is often compromised by intermittent shedding of cysts or trophozoites, as well as operator expertise. Therefore, for enhanced surveillance field-applicable, point-of-care (POC), molecular assays are needed. Our aims were to: 1) optimise the recombinase polymerase amplification (RPA) assay for the isothermal amplification of the G. duodenalis β-giardin gene from trophozoites and cysts, using published primer and probe sequences, and 2) perform a pilot field validation of RPA at a field station in a resource-poor setting, on DNA extracted from stool samples from schoolchildren in villages around Lake Albert, Uganda. Results were compared to an established laboratory small subunit ribosomal RNA (SSU rDNA) qPCR with additional testing using a specific assemblage qPCR targeting the triose phosphate isomerase (tpi) DNA regions that can distinguish G. duodenalis of two different assemblages (A and B), which are human specific.

Item Type: Article
Subjects: QX Parasitology > QX 20 Research (General)
WA Public Health > Health Problems of Special Population Groups > WA 395 Health in developing countries
WC Communicable Diseases > Tropical and Parasitic Diseases > WC 680 Tropical diseases (General)
WC Communicable Diseases > Tropical and Parasitic Diseases > WC 700 Protozoan infections (General)
Faculty: Department: Biological Sciences > Department of Tropical Disease Biology
Clinical Sciences & International Health > Clinical Sciences Department
Digital Object Identifer (DOI): https://doi.org/10.1186/s13071-020-04168-1
Depositing User: Claire McIntyre
Date Deposited: 10 Jun 2020 12:06
Last Modified: 10 Jun 2020 12:06
URI: https://archive.lstmed.ac.uk/id/eprint/14529

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