A conventional multiplex PCR for the detection of four common soil-transmitted nematodes in human feces: development and validation
https://doi.org/10.47665/tb.39.1.016
- Author:
Hassan, N.A.
1
;
Noor Badi, F.A.
2
;
Mohd-Shaharuddin, N.
1
;
Wan Yusoff, W.S.
1
;
Lim, Y.A.L.
1
;
Chua, K.H.
2
;
Sidi Omar, S.F.N.
1
;
Chang, L.Y.
3
;
Majid, H.A.
4
;
Ngui, R.
1
Author Information
1. Department of Parasitology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia
2. Department of Biomedical Science, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia
3. Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia
4. Department of Social Preventive Medicine, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia
- Publication Type:Journal Article
- Keywords:
Multiplex PCR;
soil-transmitted helminths (STHs);
intestinal nematodes;
Malaysia.
- From:Tropical Biomedicine
2022;39(No.1):135-142
- CountryMalaysia
- Language:English
-
Abstract:
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.
- Full text:8.2022my1323.pdf
