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Development of Conventional Multiplex PCR: A Rapid Technique for Simultaneous Detection of Soil-Transmitted Helminths

Soil-transmitted helminths (STHs) are the most common intestinal parasites infecting humans worldwide. STH infections are a major cause of morbidity and disability. Accurate diagnostic tools are pivotal for assessing the exact prevalence of parasitic infections. Microscopic examination and culture t...

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Autores principales: Sanprasert, Vivornpun, Kerdkaew, Ruthairat, Srirungruang, Siriporn, Charuchaibovorn, Sarit, Phadungsaksawasdi, Kobpat, Nuchprayoon, Surang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6789620/
https://www.ncbi.nlm.nih.gov/pubmed/31527459
http://dx.doi.org/10.3390/pathogens8030152
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author Sanprasert, Vivornpun
Kerdkaew, Ruthairat
Srirungruang, Siriporn
Charuchaibovorn, Sarit
Phadungsaksawasdi, Kobpat
Nuchprayoon, Surang
author_facet Sanprasert, Vivornpun
Kerdkaew, Ruthairat
Srirungruang, Siriporn
Charuchaibovorn, Sarit
Phadungsaksawasdi, Kobpat
Nuchprayoon, Surang
author_sort Sanprasert, Vivornpun
collection PubMed
description Soil-transmitted helminths (STHs) are the most common intestinal parasites infecting humans worldwide. STH infections are a major cause of morbidity and disability. Accurate diagnostic tools are pivotal for assessing the exact prevalence of parasitic infections. Microscopic examination and culture techniques have been used to observe the presence of eggs or larvae of parasites in stool samples, but they are time-consuming and have low sensitivity. Therefore, accurate, simple, and inexpensive diagnostic techniques are still required for simultaneous detection of STH infections. Although molecular-based techniques, such as real-time PCR and multiplex real-time PCR, have been developed, they are not suitable for routine diagnosis due to the requirement for expensive reagents and instruments. In this study, we established a conventional multiplex PCR for simultaneous rapid detection of Ascaris lumbricoides, Necator americanus, and Strongyloides stercoralis in stool samples. Our results show that the multiplex PCR could detect the DNA of STHs at a very low target gene concentrations (lower than 1 pg) with no cross-amplification. Multiplex PCR had five times higher sensitivity than the formalin–ethyl acetate concentration technique (FECT) in the detection of multiple infections, and two times higher for detection of S. stercoralis. However, multiplex PCR was comparable to FECT in the detection of A. lumbricoides and N. americanus. In conclusion, this method could be used as an alternative method for the detection of STHs, especially for S. stercoralis.
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spelling pubmed-67896202019-10-16 Development of Conventional Multiplex PCR: A Rapid Technique for Simultaneous Detection of Soil-Transmitted Helminths Sanprasert, Vivornpun Kerdkaew, Ruthairat Srirungruang, Siriporn Charuchaibovorn, Sarit Phadungsaksawasdi, Kobpat Nuchprayoon, Surang Pathogens Article Soil-transmitted helminths (STHs) are the most common intestinal parasites infecting humans worldwide. STH infections are a major cause of morbidity and disability. Accurate diagnostic tools are pivotal for assessing the exact prevalence of parasitic infections. Microscopic examination and culture techniques have been used to observe the presence of eggs or larvae of parasites in stool samples, but they are time-consuming and have low sensitivity. Therefore, accurate, simple, and inexpensive diagnostic techniques are still required for simultaneous detection of STH infections. Although molecular-based techniques, such as real-time PCR and multiplex real-time PCR, have been developed, they are not suitable for routine diagnosis due to the requirement for expensive reagents and instruments. In this study, we established a conventional multiplex PCR for simultaneous rapid detection of Ascaris lumbricoides, Necator americanus, and Strongyloides stercoralis in stool samples. Our results show that the multiplex PCR could detect the DNA of STHs at a very low target gene concentrations (lower than 1 pg) with no cross-amplification. Multiplex PCR had five times higher sensitivity than the formalin–ethyl acetate concentration technique (FECT) in the detection of multiple infections, and two times higher for detection of S. stercoralis. However, multiplex PCR was comparable to FECT in the detection of A. lumbricoides and N. americanus. In conclusion, this method could be used as an alternative method for the detection of STHs, especially for S. stercoralis. MDPI 2019-09-16 /pmc/articles/PMC6789620/ /pubmed/31527459 http://dx.doi.org/10.3390/pathogens8030152 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Sanprasert, Vivornpun
Kerdkaew, Ruthairat
Srirungruang, Siriporn
Charuchaibovorn, Sarit
Phadungsaksawasdi, Kobpat
Nuchprayoon, Surang
Development of Conventional Multiplex PCR: A Rapid Technique for Simultaneous Detection of Soil-Transmitted Helminths
title Development of Conventional Multiplex PCR: A Rapid Technique for Simultaneous Detection of Soil-Transmitted Helminths
title_full Development of Conventional Multiplex PCR: A Rapid Technique for Simultaneous Detection of Soil-Transmitted Helminths
title_fullStr Development of Conventional Multiplex PCR: A Rapid Technique for Simultaneous Detection of Soil-Transmitted Helminths
title_full_unstemmed Development of Conventional Multiplex PCR: A Rapid Technique for Simultaneous Detection of Soil-Transmitted Helminths
title_short Development of Conventional Multiplex PCR: A Rapid Technique for Simultaneous Detection of Soil-Transmitted Helminths
title_sort development of conventional multiplex pcr: a rapid technique for simultaneous detection of soil-transmitted helminths
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6789620/
https://www.ncbi.nlm.nih.gov/pubmed/31527459
http://dx.doi.org/10.3390/pathogens8030152
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