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Open AccessArticle

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

1
Lymphatic Filariasis and Tropical Medicine Research Unit, Chulalongkorn Medical Research Center, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
2
Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand
3
Department of Parasitology, King Chulalongkorn Memorial Hospital, Bangkok 10330, Thailand
*
Author to whom correspondence should be addressed.
Present address: Siriraj Center of Excellence for Stem Cell Research (SiSCR), Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
Pathogens 2019, 8(3), 152; https://doi.org/10.3390/pathogens8030152
Received: 14 July 2019 / Revised: 5 September 2019 / Accepted: 7 September 2019 / Published: 16 September 2019
(This article belongs to the Section Human Pathogens)
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. View Full-Text
Keywords: multiplex PCR; PCR; soil-transmitted helminths (STHs); diagnosis multiplex PCR; PCR; soil-transmitted helminths (STHs); diagnosis
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Sanprasert, V.; Kerdkaew, R.; Srirungruang, S.; Charuchaibovorn, S.; Phadungsaksawasdi, K.; Nuchprayoon, S. Development of Conventional Multiplex PCR: A Rapid Technique for Simultaneous Detection of Soil-Transmitted Helminths. Pathogens 2019, 8, 152.

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