Molecular identification of Paragonimus species by DNA pyrosequencing technology |
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| Authors: | Chairat Tantrawatpan Pewpan M Intapan Penchom Janwan Oranuch Sanpool Viraphong Lulitanond Chutatip Srichantaratsamee Witthaya Anamnart Wanchai Maleewong |
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| Institution: | 1. Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen 40002, Thailand;2. Division of Cell Biology, Department of Preclinical Sciences, Faculty of Medicine, Thammasat University, Rangsit Campus, Pathum Thani 12121, Thailand;3. Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand;4. Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand;5. Virology and Molecular Microbiology Unit, Department of Pathology, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok 14000, Thailand;6. Department of Medical Technology, School of Allied Health Sciences and Public Health, Walailak University, Thasala, Nakhon Si Thammarat 80160, Thailand |
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| Abstract: | DNA pyrosequencing for PCR amplicons is an attractive strategy for the identification of microorganisms because of its short time performance for large number of samples. In this study, the primers targeting the fragment of ITS2 region of nuclear ribosomal RNA gene were newly developed for pyrosequencing-based identification of 6 Paragonimus species, Paragonimus bangkokensis, Paragonimus harinasutai, Paragonimus heterotremus, Paragonimus macrorchis, Paragonimus siamensis and Paragonimus westermani. Pyrosequencing determination of 39 nucleotides of partial ITS2 region could discriminate 6 Paragonimus species, and could also detect intra-species genetic variation of P. macrorchis. This DNA pyrosequencing-based identification can be a valuable tool to improve species-level identification of Paragonimus in the endemic areas. |
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