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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doaj-2947bf63e1d647029d59d18ee27ab4432020-11-24T21:26:40ZengMDPI AGPathogens2076-08172019-09-018315210.3390/pathogens8030152pathogens8030152Development of Conventional Multiplex PCR: A Rapid Technique for Simultaneous Detection of Soil-Transmitted HelminthsVivornpun Sanprasert0Ruthairat Kerdkaew1Siriporn Srirungruang2Sarit Charuchaibovorn3Kobpat Phadungsaksawasdi4Surang Nuchprayoon5Lymphatic Filariasis and Tropical Medicine Research Unit, Chulalongkorn Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, ThailandLymphatic Filariasis and Tropical Medicine Research Unit, Chulalongkorn Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, ThailandLymphatic Filariasis and Tropical Medicine Research Unit, Chulalongkorn Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, ThailandLymphatic Filariasis and Tropical Medicine Research Unit, Chulalongkorn Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, ThailandDepartment of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, ThailandLymphatic Filariasis and Tropical Medicine Research Unit, Chulalongkorn Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, ThailandSoil-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 <i>Ascaris lumbricoides</i>, <i>Necator americanus</i>, and <i>Strongyloides stercoralis</i> 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 <i>S. stercoralis</i>. However, multiplex PCR was comparable to FECT in the detection of <i>A. lumbricoides</i> and <i>N. americanus</i>. In conclusion, this method could be used as an alternative method for the detection of STHs, especially for <i>S. stercoralis</i>.https://www.mdpi.com/2076-0817/8/3/152multiplex PCRPCRsoil-transmitted helminths (STHs)diagnosis |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Vivornpun Sanprasert Ruthairat Kerdkaew Siriporn Srirungruang Sarit Charuchaibovorn Kobpat Phadungsaksawasdi Surang Nuchprayoon |
spellingShingle |
Vivornpun Sanprasert Ruthairat Kerdkaew Siriporn Srirungruang Sarit Charuchaibovorn Kobpat Phadungsaksawasdi Surang Nuchprayoon Development of Conventional Multiplex PCR: A Rapid Technique for Simultaneous Detection of Soil-Transmitted Helminths Pathogens multiplex PCR PCR soil-transmitted helminths (STHs) diagnosis |
author_facet |
Vivornpun Sanprasert Ruthairat Kerdkaew Siriporn Srirungruang Sarit Charuchaibovorn Kobpat Phadungsaksawasdi Surang Nuchprayoon |
author_sort |
Vivornpun Sanprasert |
title |
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_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_sort |
development of conventional multiplex pcr: a rapid technique for simultaneous detection of soil-transmitted helminths |
publisher |
MDPI AG |
series |
Pathogens |
issn |
2076-0817 |
publishDate |
2019-09-01 |
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 <i>Ascaris lumbricoides</i>, <i>Necator americanus</i>, and <i>Strongyloides stercoralis</i> 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 <i>S. stercoralis</i>. However, multiplex PCR was comparable to FECT in the detection of <i>A. lumbricoides</i> and <i>N. americanus</i>. In conclusion, this method could be used as an alternative method for the detection of STHs, especially for <i>S. stercoralis</i>. |
topic |
multiplex PCR PCR soil-transmitted helminths (STHs) diagnosis |
url |
https://www.mdpi.com/2076-0817/8/3/152 |
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