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Pathogens. 2019 Sep 16;8(3). pii: E152. doi: 10.3390/pathogens8030152.

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

Author information

1
Lymphatic Filariasis and Tropical Medicine Research Unit, Chulalongkorn Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. vivornpun@chula.md.
2
Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. vivornpun@chula.md.
3
Lymphatic Filariasis and Tropical Medicine Research Unit, Chulalongkorn Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. joruthairat@gmail.com.
4
Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. joruthairat@gmail.com.
5
Lymphatic Filariasis and Tropical Medicine Research Unit, Chulalongkorn Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. lingnooy55@windowslive.com.
6
Department of Parasitology, King Chulalongkorn Memorial Hospital, Bangkok 10330, Thailand. lingnooy55@windowslive.com.
7
Lymphatic Filariasis and Tropical Medicine Research Unit, Chulalongkorn Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. sarit.src@gmail.com.
8
Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. kobpat22@hotmail.com.
9
Department of Parasitology, King Chulalongkorn Memorial Hospital, Bangkok 10330, Thailand. kobpat22@hotmail.com.
10
Lymphatic Filariasis and Tropical Medicine Research Unit, Chulalongkorn Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. fmedstt@gmail.com.
11
Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand. fmedstt@gmail.com.

Abstract

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.

KEYWORDS:

PCR; diagnosis; multiplex PCR; soil-transmitted helminths (STHs)

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