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Development and evaluation of a loop-mediated isothermal amplification (LAMP) diagnostic test for detection of whipworm, Trichuris trichiura, in faecal samples

Published online by Cambridge University Press:  02 April 2020

M.G. Ngari ,
I.N. Mwangi ,
M.P. Njoroge ,
J. Kinyua ,
F.A. Osuna ,
B.M. Kimeu ,
P.W. Okanya  and
E.L. Agola Open the ORCID record for E.L. Agola [Opens in a new window]
M.G. Ngari
Affiliation:
Department of Biochemistry, Jomo Kenyatta University of Agriculture and Technology, PO Box 62000, 00200Nairobi, Kenya
I.N. Mwangi
Affiliation:
Centre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), PO Box 54840, 00200Nairobi, Kenya
M.P. Njoroge
Affiliation:
Department of Health, PO Box 24, Kirinyaga County, Kerugoya, Kenya
J. Kinyua
Affiliation:
Department of Biochemistry, Jomo Kenyatta University of Agriculture and Technology, PO Box 62000, 00200Nairobi, Kenya
F.A. Osuna
Affiliation:
Centre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), PO Box 54840, 00200Nairobi, Kenya
B.M. Kimeu
Affiliation:
Department of Biochemistry and Biotechnology, Technical University of Kenya, PO Box 52428, 00200Nairobi, Kenya
P.W. Okanya
Affiliation:
Department of Biochemistry and Biotechnology, Technical University of Kenya, PO Box 52428, 00200Nairobi, Kenya
E.L. Agola*
Affiliation:
Centre for Biotechnology Research and Development, Kenya Medical Research Institute (KEMRI), PO Box 54840, 00200Nairobi, Kenya Department of Biochemistry and Biotechnology, Technical University of Kenya, PO Box 52428, 00200Nairobi, Kenya
*
Author for correspondence: E.L. Agola, E-mail: [email protected]
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Abstract

Whipworm infection or trichuriasis caused by Trichuris trichiura is of major public health concern in sub-Saharan Africa, particularly among pre-school and school-going children. It is among the neglected tropical diseases targeted for elimination through mass drug administration (MDA). One of the outcomes of MDA is a rapid decline in levels of infection intensity, making it difficult to monitor effectiveness of control measures using the conventional Kato–Katz procedure, which relies on the microscopic detection of parasite ova in faecal samples. In the present study, a loop-mediated isothermal amplification (LAMP) test was developed for the detection of T. trichiura infection in faecal samples. LAMP technology offers greater sensitivity and specificity than the microscopy-based tests. A set of four specific primers targeting the internal transcribed spacer 2 region of the ribosomal DNA were designed using Primer Explorer software. DNA was extracted from faecal samples using the alkaline lysis method (HotSHOT) and the LAMP reaction performed at 63°C for 1 h. The amplicons were visualized by both gel electrophoresis and with the naked eye following staining with SYBR green dye. Sensitivity and specificity tests were determined using the standard Kato–Katz diagnostic procedure as a reference test. The developed LAMP assay reliably detected T. trichiura DNA in faecal samples, with a specificity and sensitivity of 88% and 77%, respectively. No cross-reactivity was observed with several common helminth parasites. The developed LAMP assay is an appropriate diagnostic method for the detection of T. trichiura DNA in human faecal samples due to its simplicity, low cost, high sensitivity and specificity.

Keywords

Kato–KatzLAMP assayTrichuris trichiuratrichuriasisdiagnosisITS-2Kenya
Type
Research Paper
Information
Journal of Helminthology , Volume 94 , 2020 , e142
Copyright
Copyright © The Author(s), 2020. Published by Cambridge University Press

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