Rapid detection and identification of human hookworm infections through high resolution melting (HRM) analysis.

<h4>Background</h4>Hookworm infections are still endemic in low and middle income tropical countries with greater impact on the socioeconomic and public health of the bottom billion of the world's poorest people. In this study, a real-time polymerase chain reaction (PCR) coupled wit...

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Autores principales:	Romano Ngui, Yvonne A L Lim, Kek Heng Chua
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/56dd6be5930c47579fdbf581a7a830b5
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spelling	oai:doaj.org-article:56dd6be5930c47579fdbf581a7a830b52021-11-18T07:10:54ZRapid detection and identification of human hookworm infections through high resolution melting (HRM) analysis.1932-620310.1371/journal.pone.0041996https://doaj.org/article/56dd6be5930c47579fdbf581a7a830b52012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22844538/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Hookworm infections are still endemic in low and middle income tropical countries with greater impact on the socioeconomic and public health of the bottom billion of the world's poorest people. In this study, a real-time polymerase chain reaction (PCR) coupled with high resolution melting-curve (HRM) analysis was evaluated for an accurate, rapid and sensitive tool for species identification focusing on the five human hookworm species.<h4>Methods</h4>Real-time PCR coupled with HRM analysis targeting the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA as the genetic marker was used to identify and distinguish hookworm species in human samples. Unique and distinct characteristics of HRM patterns were produced for each of the five hookworm species. The melting curves were characterized by peaks of 79.24±0.05°C and 83.00±0.04°C for Necator americanus, 79.12±0.10°C for Ancylostoma duodenale, 79.40±0.10°C for Ancylostoma ceylanicum, 79.63±0.05°C for Ancylostoma caninum and 79.70±0.14°C for Ancylostoma braziliense. An evaluation of the method's sensitivity and specificity revealed that this assay was able to detect as low as 0.01 ng/µl hookworm DNA and amplification was only recorded for hookworm positive samples.<h4>Conclusion</h4>The HRM assay developed in this study is a rapid and straightforward method for the diagnosis, identification and discrimination of five human hookworms. This assay is simple compared to other probe-based genotyping methods as it does not require multiplexing, DNA sequencing or post-PCR processing. Therefore, this method offers a new alternative for rapid detection of human hookworm species.Romano NguiYvonne A L LimKek Heng ChuaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 7, p e41996 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Romano Ngui Yvonne A L Lim Kek Heng Chua Rapid detection and identification of human hookworm infections through high resolution melting (HRM) analysis.
description	<h4>Background</h4>Hookworm infections are still endemic in low and middle income tropical countries with greater impact on the socioeconomic and public health of the bottom billion of the world's poorest people. In this study, a real-time polymerase chain reaction (PCR) coupled with high resolution melting-curve (HRM) analysis was evaluated for an accurate, rapid and sensitive tool for species identification focusing on the five human hookworm species.<h4>Methods</h4>Real-time PCR coupled with HRM analysis targeting the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA as the genetic marker was used to identify and distinguish hookworm species in human samples. Unique and distinct characteristics of HRM patterns were produced for each of the five hookworm species. The melting curves were characterized by peaks of 79.24±0.05°C and 83.00±0.04°C for Necator americanus, 79.12±0.10°C for Ancylostoma duodenale, 79.40±0.10°C for Ancylostoma ceylanicum, 79.63±0.05°C for Ancylostoma caninum and 79.70±0.14°C for Ancylostoma braziliense. An evaluation of the method's sensitivity and specificity revealed that this assay was able to detect as low as 0.01 ng/µl hookworm DNA and amplification was only recorded for hookworm positive samples.<h4>Conclusion</h4>The HRM assay developed in this study is a rapid and straightforward method for the diagnosis, identification and discrimination of five human hookworms. This assay is simple compared to other probe-based genotyping methods as it does not require multiplexing, DNA sequencing or post-PCR processing. Therefore, this method offers a new alternative for rapid detection of human hookworm species.
format	article
author	Romano Ngui Yvonne A L Lim Kek Heng Chua
author_facet	Romano Ngui Yvonne A L Lim Kek Heng Chua
author_sort	Romano Ngui
title	Rapid detection and identification of human hookworm infections through high resolution melting (HRM) analysis.
title_short	Rapid detection and identification of human hookworm infections through high resolution melting (HRM) analysis.
title_full	Rapid detection and identification of human hookworm infections through high resolution melting (HRM) analysis.
title_fullStr	Rapid detection and identification of human hookworm infections through high resolution melting (HRM) analysis.
title_full_unstemmed	Rapid detection and identification of human hookworm infections through high resolution melting (HRM) analysis.
title_sort	rapid detection and identification of human hookworm infections through high resolution melting (hrm) analysis.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/56dd6be5930c47579fdbf581a7a830b5
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Rapid detection and identification of human hookworm infections through high resolution melting (HRM) analysis.

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