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

Sandra J. Molina-Gonzalez; Sandra J. Molina-Gonzalez; Tapan Bhattacharyya; Tapan Bhattacharyya; Hajri R. Alshehri; Hajri R. Alshehri; Kate Poulton; Kate Poulton; Stephen Allen; Michael A. Miles; Michael A. Miles; Moses Arianitwe; Edridah M. Tukahebwa; Bonnie Webster; Bonnie Webster; Russell Stothard; Amaya L. Bustinduy; Amaya L. Bustinduy

doi:10.1186/s13071-020-04168-1

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

Sandra J. Molina-Gonzalez, Sandra J. Molina-Gonzalez, Tapan Bhattacharyya, Tapan Bhattacharyya, Hajri R. Alshehri, Hajri R. Alshehri, Kate Poulton, Kate Poulton, Stephen Allen, Michael A. Miles, Michael A. Miles, Moses Arianitwe, Edridah M. Tukahebwa, Bonnie Webster, Bonnie Webster, Russell Stothard, Amaya L. Bustinduy, Amaya L. Bustinduy

Research output: Contribution to journal › Article › peer-review

12 Citations (Scopus)

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.

Original language	English
Article number	289
Journal	Parasites and Vectors
Volume	13
Issue number	1
DOIs	https://doi.org/10.1186/s13071-020-04168-1
Publication status	Published - 6 Jun 2020

Keywords

Assemblage typing
Epidemiology
Giardia duodenalis
Giardia intestinalis
Giardia lamblia
Giardiasis
Point-of-care
Recombinase polymerase amplification
Uganda

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Molina-Gonzalez, S. J., Molina-Gonzalez, S. J., Bhattacharyya, T., Bhattacharyya, T., Alshehri, H. R., Alshehri, H. R., Poulton, K., Poulton, K., Allen, S., Miles, M. A., Miles, M. A., Arianitwe, M., Tukahebwa, E. M., Webster, B., Webster, B., Stothard, R., Bustinduy, A. L., & Bustinduy, A. L. (2020). Application of a Recombinase Polymerase Amplification (RPA) assay and pilot field testing for Giardia duodenalis at Lake Albert, Uganda. Parasites and Vectors, 13(1), Article 289. https://doi.org/10.1186/s13071-020-04168-1

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

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.",

keywords = "Assemblage typing, Epidemiology, Giardia duodenalis, Giardia intestinalis, Giardia lamblia, Giardiasis, Point-of-care, Recombinase polymerase amplification, Uganda",

author = "Molina-Gonzalez, \{Sandra J.\} and Molina-Gonzalez, \{Sandra J.\} and Tapan Bhattacharyya and Tapan Bhattacharyya and Alshehri, \{Hajri R.\} and Alshehri, \{Hajri R.\} and Kate Poulton and Kate Poulton and Stephen Allen and Miles, \{Michael A.\} and Miles, \{Michael A.\} and Moses Arianitwe and Tukahebwa, \{Edridah M.\} and Bonnie Webster and Bonnie Webster and Russell Stothard and Bustinduy, \{Amaya L.\} and Bustinduy, \{Amaya L.\}",

year = "2020",

month = jun,

day = "6",

doi = "10.1186/s13071-020-04168-1",

language = "English",

volume = "13",

journal = "Parasites and Vectors",

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publisher = "BioMed Central Ltd",

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Molina-Gonzalez, SJ, Molina-Gonzalez, SJ, Bhattacharyya, T, Bhattacharyya, T, Alshehri, HR, Alshehri, HR, Poulton, K, Poulton, K, Allen, S, Miles, MA, Miles, MA, Arianitwe, M, Tukahebwa, EM, Webster, B, Webster, B, Stothard, R, Bustinduy, AL & Bustinduy, AL 2020, 'Application of a Recombinase Polymerase Amplification (RPA) assay and pilot field testing for Giardia duodenalis at Lake Albert, Uganda.', Parasites and Vectors, vol. 13, no. 1, 289. https://doi.org/10.1186/s13071-020-04168-1

TY - JOUR

T1 - Application of a Recombinase Polymerase Amplification (RPA) assay and pilot field testing for Giardia duodenalis at Lake Albert, Uganda.

AU - Molina-Gonzalez, Sandra J.

AU - Bhattacharyya, Tapan

AU - Alshehri, Hajri R.

AU - Poulton, Kate

AU - Allen, Stephen

AU - Miles, Michael A.

AU - Arianitwe, Moses

AU - Tukahebwa, Edridah M.

AU - Webster, Bonnie

AU - Stothard, Russell

AU - Bustinduy, Amaya L.

PY - 2020/6/6

Y1 - 2020/6/6

N2 - 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.

AB - 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.

KW - Assemblage typing

KW - Epidemiology

KW - Giardia duodenalis

KW - Giardia intestinalis

KW - Giardia lamblia

KW - Giardiasis

KW - Point-of-care

KW - Recombinase polymerase amplification

KW - Uganda

U2 - 10.1186/s13071-020-04168-1

DO - 10.1186/s13071-020-04168-1

M3 - Article

SN - 1756-3305

VL - 13

JO - Parasites and Vectors

JF - Parasites and Vectors

IS - 1

M1 - 289

ER -

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

Abstract

Keywords

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