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Please use this identifier to cite or link to this item: http://hdl.handle.net/123456789/11978

Title: An isothermal DNA amplification method for detection of Onchocerca volvulus infection in skin biopsies
Authors: Lagatie, Ole
Merino, Michelle
Debrah, Linda Batsa
Debrah, Alexander Y.
Stuyver, Lieven J.
Keywords: Onchocerca volvulus
River blindness
Onchocerciasis
Skin biopsy
DNA
cox1
LAMP
Isothermal
Diagnostic
Issue Date: Dec-2016
Publisher: Parasites & Vectors
Citation: Parasites & Vectors 9(1)
Abstract: Background Diagnostic procedures for the diagnosis of infection with the nematode parasite Onchocerca volvulus are currently based on the microscopic detection of microfilariae in skin biopsies. Alternative approaches based on amplification of parasitic DNA in these skin biopsies are currently being explored. Mostly this is based on the detection of the O-150 repeat sequence using PCR based techniques. Methods An isothermal, loop-mediated amplification method has been designed using the mitochondrial O. volvulus cox1 gene as a target. ResultsAnalysis of dilution series of synthetic DNA containing the targeted sequence show a non-linear dose-response curve, as is usually the case for isothermal amplification methods. Evaluation of cross-reactivity with the heterologous sequence from the closely related parasites Wuchereria bancrofti, Loa loa and Brugia malayi demonstrated strong specificity, as none of these sequences was amplified. The assay however amplified both O. volvulus and O. ochengi DNA, but with a different melting point that can be used to discriminate between the species. Evaluation of this assay in a set of skin snip biopsies collected in an endemic area in Ghana showed a high correlation with O-150 qPCR and also demonstrated a similar sensitivity. Compared to qPCR, LAMP had a sensitivity of 88.2% and a specificity of 99.2%. Conclusions We have developed a sensitive and specific loop-mediated amplification method for detection of O. volvulus DNA in skin biopsies that is capable of providing results within 30 min.
Description: This article is published in Parasites & Vectors and also available at DOI: 10.1186/s13071-016-1913-7
URI: 10.1186/s13071-016-1913-7
http://hdl.handle.net/123456789/11978
Appears in Collections:College of Health Sciences

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