Cover Image

A conventional multiplex PCR for the detection of four common soil-transmitted nematodes in human feces : development and validation

Soil-transmitted helminth (STH) infections, mainly caused by Ascaris lumbricoides, Trichuris trichiura, and hookworms, are among the most common intestinal parasites that infect humans. The infections are widely distributed throughout tropical and subtropical countries, including Malaysia, particul...

Full description

Saved in:
Bibliographic Details
Main Authors: Hassan, N.A., Noor Badi, F.A, Mohd-Shaharuddin, N., Wan Yusoff, W. S, Lim, Y.A.L., Chua, K.H., Sidi Omar, S.F.N, Chang, L.Y., Majid, H. A, Romano, Ngui
Format: Article
Language:English
Published: The Malaysian Society of Parasitology and Tropical Medicine (MSPTM) 2022
Subjects:
QR Microbiology
Online Access:http://ir.unimas.my/id/eprint/42131/1/A%20conventional.pdf
http://ir.unimas.my/id/eprint/42131/
https://msptm.org/vol-39-no-1-march/
https://doi.org/10.47665/tb.39.1.016
Tags: Add Tag
No Tags, Be the first to tag this record!
id my.unimas.ir.42131
record_format eprints
spelling my.unimas.ir.421312023-07-05T02:08:40Z http://ir.unimas.my/id/eprint/42131/ A conventional multiplex PCR for the detection of four common soil-transmitted nematodes in human feces : development and validation Hassan, N.A. Noor Badi, F.A Mohd-Shaharuddin, N. Wan Yusoff, W. S Lim, Y.A.L. Chua, K.H. Sidi Omar, S.F.N Chang, L.Y. Majid, H. A Romano, Ngui QR Microbiology Soil-transmitted helminth (STH) infections, mainly caused by Ascaris lumbricoides, Trichuris trichiura, and hookworms, are among the most common intestinal parasites that infect humans. The infections are widely distributed throughout tropical and subtropical countries, including Malaysia, particularly in underprivileged communities. Microscopic and culture techniques have been used as a gold standard for diagnostic techniques. However, these methods yield low sensitivity and specificity, laborious and time-consuming. Therefore, simple, rapid, and accurate alternative methods are needed for the simultaneous detection of STH infections. Although advanced technologies such as real-time multiplex PCR have been established, the use of this technique as a routine diagnostic is limited due to the high cost of the instrument. Therefore, a single-round multiplex conventional PCR assay for rapid detection of four STH species in the fecal sample was developed in this study. To perform the single-round multiplex PCR, each pair of species-specific primers was selected from target genes, including Ancylostoma duodenale (Internal Transcribed Spacer 2; accession No. AJ001594; 156 base pair), Necator americanus (ITS 2; accession No. AJ001599; 225 base pair), Ascaris lumbricoides (Internal Transcribed Spacer 1; accession No. AJ000895; 334 base pair) and Trichuris triciura (partial ITS 1, 5.8s rRNA and partial ITS 2; accession No. AM992981; 518 base pair). The results showed that the newly designed primers could detect the DNA of STH at low concentrations (0.001 ng/μl) with no cross-amplification with other species. This assay enables the differentiation of single infections as well as mixed infections. It could be used as an alternative and is a convenient method for the detection of STHs, especially for the differentiation of N. americanus and A. duodenale. The Malaysian Society of Parasitology and Tropical Medicine (MSPTM) 2022 Article PeerReviewed text en http://ir.unimas.my/id/eprint/42131/1/A%20conventional.pdf Hassan, N.A. and Noor Badi, F.A and Mohd-Shaharuddin, N. and Wan Yusoff, W. S and Lim, Y.A.L. and Chua, K.H. and Sidi Omar, S.F.N and Chang, L.Y. and Majid, H. A and Romano, Ngui (2022) A conventional multiplex PCR for the detection of four common soil-transmitted nematodes in human feces : development and validation. Tropical Biomedicine, 39 (1). pp. 135-143. ISSN 2521-9855. https://msptm.org/vol-39-no-1-march/ https://doi.org/10.47665/tb.39.1.016
institution Universiti Malaysia Sarawak
building Centre for Academic Information Services (CAIS)
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaysia Sarawak
content_source UNIMAS Institutional Repository
url_provider http://ir.unimas.my/
language English
topic QR Microbiology
spellingShingle QR Microbiology
Hassan, N.A.
Noor Badi, F.A
Mohd-Shaharuddin, N.
Wan Yusoff, W. S
Lim, Y.A.L.
Chua, K.H.
Sidi Omar, S.F.N
Chang, L.Y.
Majid, H. A
Romano, Ngui
A conventional multiplex PCR for the detection of four common soil-transmitted nematodes in human feces : development and validation
description Soil-transmitted helminth (STH) infections, mainly caused by Ascaris lumbricoides, Trichuris trichiura, and hookworms, are among the most common intestinal parasites that infect humans. The infections are widely distributed throughout tropical and subtropical countries, including Malaysia, particularly in underprivileged communities. Microscopic and culture techniques have been used as a gold standard for diagnostic techniques. However, these methods yield low sensitivity and specificity, laborious and time-consuming. Therefore, simple, rapid, and accurate alternative methods are needed for the simultaneous detection of STH infections. Although advanced technologies such as real-time multiplex PCR have been established, the use of this technique as a routine diagnostic is limited due to the high cost of the instrument. Therefore, a single-round multiplex conventional PCR assay for rapid detection of four STH species in the fecal sample was developed in this study. To perform the single-round multiplex PCR, each pair of species-specific primers was selected from target genes, including Ancylostoma duodenale (Internal Transcribed Spacer 2; accession No. AJ001594; 156 base pair), Necator americanus (ITS 2; accession No. AJ001599; 225 base pair), Ascaris lumbricoides (Internal Transcribed Spacer 1; accession No. AJ000895; 334 base pair) and Trichuris triciura (partial ITS 1, 5.8s rRNA and partial ITS 2; accession No. AM992981; 518 base pair). The results showed that the newly designed primers could detect the DNA of STH at low concentrations (0.001 ng/μl) with no cross-amplification with other species. This assay enables the differentiation of single infections as well as mixed infections. It could be used as an alternative and is a convenient method for the detection of STHs, especially for the differentiation of N. americanus and A. duodenale.
format Article
author Hassan, N.A.
Noor Badi, F.A
Mohd-Shaharuddin, N.
Wan Yusoff, W. S
Lim, Y.A.L.
Chua, K.H.
Sidi Omar, S.F.N
Chang, L.Y.
Majid, H. A
Romano, Ngui
author_facet Hassan, N.A.
Noor Badi, F.A
Mohd-Shaharuddin, N.
Wan Yusoff, W. S
Lim, Y.A.L.
Chua, K.H.
Sidi Omar, S.F.N
Chang, L.Y.
Majid, H. A
Romano, Ngui
author_sort Hassan, N.A.
title A conventional multiplex PCR for the detection of four common soil-transmitted nematodes in human feces : development and validation
title_short A conventional multiplex PCR for the detection of four common soil-transmitted nematodes in human feces : development and validation
title_full A conventional multiplex PCR for the detection of four common soil-transmitted nematodes in human feces : development and validation
title_fullStr A conventional multiplex PCR for the detection of four common soil-transmitted nematodes in human feces : development and validation
title_full_unstemmed A conventional multiplex PCR for the detection of four common soil-transmitted nematodes in human feces : development and validation
title_sort conventional multiplex pcr for the detection of four common soil-transmitted nematodes in human feces : development and validation
publisher The Malaysian Society of Parasitology and Tropical Medicine (MSPTM)
publishDate 2022
url http://ir.unimas.my/id/eprint/42131/1/A%20conventional.pdf
http://ir.unimas.my/id/eprint/42131/
https://msptm.org/vol-39-no-1-march/
https://doi.org/10.47665/tb.39.1.016
_version_ 1770555415154655232
score 13.211869

Similar Items