Parasites, Hosts and Diseases

"Pham Ngoc Doanh"

Original Articles

Molecular Differentiation of Opisthorchis viverrini and Clonorchis sinensis Eggs by Multiplex Real-Time PCR with High Resolution Melting Analysis: Worasak Kaewkong, Pewpan M. Intapan, Oranuch Sanpool, Penchom Janwan, Tongjit Thanchomnang, Porntip Laummaunwai, Viraphong Lulitanond, Pham Ngoc Doanh, Wanchai Maleewong; Korean J Parasitol 2013;51(6):689-694.
Published online December 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.6.689

Abstract
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Opisthorchis viverrini and Clonorchis sinensis are parasites known to be carcinogenic and causative agents of cholangiocarcinoma in Asia. The standard method for diagnosis for those parasite infections is stool examination to detect parasite eggs. However, the method has low sensitivity, and eggs of O. viverrini and C. sinensis are difficult to distinguish from each other and from those of some other trematodes. Here, we report a multiplex real-time PCR coupled with high resolution melting (HRM) analysis for the differentiation of O. viverrini and C. sinensis eggs in fecal samples. Using 2 pairs of species-specific primers, DNA sequences from a portion of the mitochondrial NADH dehydrogenase subunit 2 (nad 2) gene, were amplified to generate 209 and 165 bp products for O. viverrini and C. sinensis, respectively. The distinct characteristics of HRM patterns were analyzed, and the melting temperatures peaked at 82.4±0.09℃ and 85.9±0.08℃ for O. viverrini and C. sinensis, respectively. This technique was able to detect as few as 1 egg of O. viverrini and 2 eggs of C. sinensis in a 150 mg fecal sample, which is equivalent to 7 and 14 eggs per gram of feces, respectively. The method is species-specific, rapid, simple, and does not require fluorescent probes or post-PCR processing for discrimination of eggs of the 2 species. It offers a new tool for differentiation and detection of Asian liver fluke infections in stool specimens.

Citations

Citations to this article as recorded by

Multiparasitism in Spain in a Korean Visiting Friends and Relatives: Case Report and Review of Imported Cases of Clonorchis sinensis in Europe
Paola Cociancic, Jacklyn Comas, J. Guillermo Esteban
Clinical Case Reports.2025;[Epub] CrossRef
Study of the population genetic structure of Opisthorchis-like eggs in northern Thailand using mitochondrial genes
Picha Suwannahitatorn, Mathirut Mungthin, Ittisak Subrungruang, Lakhanawan Charoensuk, Nithikoon Aksorn, Saiwasan Buathong, Krystyna Cwiklinski
PLOS Neglected Tropical Diseases.2024; 18(8): e0012445. CrossRef
An Approach for Egg Parasite Classification Based on Ensemble Deep Learning
Narut Butploy, Wanida Kanarkard, Pewpan M. Intapan, Oranuch Sanpool
Journal of Advanced Computational Intelligence and Intelligent Informatics.2023; 27(6): 1113. CrossRef
Fish and Food-Fatale: Food-borne Trematode Opisthorchis viverrini and Cholangiocarcinoma
S. Tan, M. Machrumnizar
Helminthologia.2023; 60(4): 287. CrossRef
Are Melanoides tuberculata and Tarebia granifera (Gastropoda, Thiaridae), suitable first intermediate hosts of Clonorchis sinensis in Vietnam?
Hung Manh Nguyen, Hien Hoang Van, Loan Thi Ho, Yulia V. Tatonova, Henry Madsen, Xiao-Nong Zhou
PLOS Neglected Tropical Diseases.2021; 15(1): e0009093. CrossRef
Clonorchis sinensis and clonorchiasis
Byoung-Kuk Na, Jhang Ho Pak, Sung-Jong Hong
Acta Tropica.2020; 203: 105309. CrossRef
Serodiagnostic antigens of Clonorchis sinensis identified and evaluated by high-throughput proteogenomics
Pyo Yun Cho, Ji-Yun Lee, Tae Im Kim, Jin-Ho Song, Sung-Jong Hong, Won Gi Yoo, Takafumi Tsuboi, Kwon-Soo Ha, Jae-Wan Jung, Satoru Takeo, Eun-Taek Han, Banchob Sripa, Sung-Tae Hong, Jong-Yil Chai, Ho-Woo Nam, Jhang Ho Pak, Tong-Soo Kim, Krystyna Cwiklinski
PLOS Neglected Tropical Diseases.2020; 14(12): e0008998. CrossRef
Clonorchiasis sinensis detected by laparoscopic exploration of biliary tracts in two patients with obstructive jaundice
Xialei Liu, Genglong Zhu, Chaonong Cai, Zhiyue Lv, Jian Li
BMC Infectious Diseases.2019;[Epub] CrossRef
Development of Conventional Multiplex PCR: A Rapid Technique for Simultaneous Detection of Soil-Transmitted Helminths
Vivornpun Sanprasert, Ruthairat Kerdkaew, Siriporn Srirungruang, Sarit Charuchaibovorn, Kobpat Phadungsaksawasdi, Surang Nuchprayoon
Pathogens.2019; 8(3): 152. CrossRef
Performance evaluation of existing immunoassays for Clonorchis sinensis infection in China
Hong-Mei Li, Men-Bao Qian, Yi-Chao Yang, Zhi-Hua Jiang, Kang Wei, Jia-Xu Chen, Jun-Hu Chen, Ying-Dan Chen, Xiao-Nong Zhou
Parasites & Vectors.2018;[Epub] CrossRef
Molecular characterization of the unique Mesostephanus appendiculatus (Trematoda: Cyathocotylidae) by small ribosomal RNA from Egypt
Nasr M. El-Bahy, Eman K. Bazh, Shimaa S. Sorour, Nagwa M. Elhawary
Parasitology Research.2017; 116(4): 1129. CrossRef
LAMPhimerus: A novel LAMP assay for detecting Amphimerus sp. DNA in human stool samples
William Cevallos, Pedro Fernández-Soto, Manuel Calvopiña, Cristina Fontecha-Cuenca, Hiromu Sugiyama, Megumi Sato, Julio López Abán, Belén Vicente, Antonio Muro, David Blair
PLOS Neglected Tropical Diseases.2017; 11(6): e0005672. CrossRef
Molecular discrimination of Opisthorchis-like eggs from residents in a rural community of central Thailand
Saiwasan Buathong, Saovanee Leelayoova, Mathirut Mungthin, Toon Ruang-areerate, Tawee Naaglor, Picha Suwannahitatorn, Phunlerd Piyaraj, Paanjit Taamasri, Peerapan Tan-ariya, Edoardo Pozio
PLOS Neglected Tropical Diseases.2017; 11(11): e0006030. CrossRef
Current status ofClonorchis sinensisand clonorchiasis in China
De-Hua Lai, Xiao-Kun Hong, Bi-Xiu Su, Chi Liang, Geoff Hide, Xiaoli Zhang, Xinbing Yu, Zhao-Rong Lun
Transactions of The Royal Society of Tropical Medicine and Hygiene.2016; 110(1): 21. CrossRef

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Paragonimus paishuihoensis Metacercariae in Freshwater Crabs, Potamon lipkei, in Vientiane Province, Lao PDR: Shigehisa Habe, Pham Ngoc Doanh, Shinichiro Yahiro, Nanthasane Vannavong, Hubert Barennes, Peter Odermatt, Gilles Dreyfuss, Yoichiro Horii, Yukifumi Nawa; Korean J Parasitol 2013;51(6):683-687.
Published online December 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.6.683

Abstract
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Among Paragonimus species, P. paishuihoensis is one of the most mysterious and poorly understood species. Metacercariae are characterized by having a unique dendritically branched excretory bladder. However, the morphology of the adult worm remains unknown. To date, metacercariae of this species have been reported only in China and Thailand. In this study, we first found P. paishuihoensis metacercariae in freshwater crabs, Potamon lipkei, in Hinheub District, Vientiane, Lao PDR, with a prevalence of 77.7% and the average intensity of 10.3 (range 1-28) metacercariae per crab. The molecular data based on ITS2 and CO1 markers indicated that P. paishuihoensis from Laos and Thailand were almost completely identical and were close to members of the Paragonimus bangkokensis/Paragonimus harinasutai complex. Attempts to infect experimental animals (cats, dogs, and rats) with P. paishuihoensis were unsuccessful, suggesting that these animals might be unsuitable definitive hosts for the species. Further studies are necessary to elucidate the taxonomic status and life cycle of P. paishuihoensis.

Citations

Citations to this article as recorded by

Lung flukes of the genusParagonimus: ancient and re-emerging pathogens
David Blair
Parasitology.2022; 149(10): 1286. CrossRef
Paragonimus and paragonimiasis in Asia: An update
Ayako Yoshida, Pham Ngoc Doanh, Haruhiko Maruyama
Acta Tropica.2019; 199: 105074. CrossRef
Morphological and molecular identification of a lung fluke, Paragonimus macrorchis (Trematoda, Paragonimidae), found in central Lao PDR and its molecular phylogenetic status in the genus Paragonimus
Oranuch Sanpool, Pewpan Maleewong Intapan, Tongjit Thanchomnang, Penchom Janwan, Sakhone Laymanivong, Hiromu Sugiyama, Wanchai Maleewong
Parasitology International.2015; 64(6): 513. CrossRef

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120 Download
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Morphological and Molecular Characterization of Lymnaeid Snails and Their Potential Role in Transmission of Fasciola spp. in Vietnam: Bui Thi Dung, Pham Ngoc Doanh, Dang Tat The, Ho Thi Loan, Bertrand Losson, Yannick Caron; Korean J Parasitol 2013;51(6):657-662.
Published online December 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.6.657

Abstract
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Freshwater snails of the family Lymnaeidae play an important role in the transmission of fascioliasis worldwide. In Vietnam, 2 common lymnaeid species, Lymnaea swinhoei and Lymnaea viridis, can be recognized on the basis of morphology, and a third species, Lymnaea sp., is known to exist. Recent studies have raised controversy about their role in transmission of Fasciola spp. because of confusion in identification of the snail hosts. The aim of this study is, therefore, to clarify the identities of lymnaeid snails in Vietnam by a combination of morphological and molecular approaches. The molecular analyses using the second internal transcribed spacer (ITS2) of the nuclear ribosomal DNA clearly showed that lymnaeids in Vietnam include 3 species, Austropeplea viridis (morphologically identified as L. viridis), Radix auricularia (morphologically identified as L. swinhoei) and Radix rubiginosa (morphologically identified as Lymnaea sp.). R. rubiginosa is a new record for Vietnam. Among them, only A. viridis was found to be infected with Fasciola spp. These results provide a new insight into lymnaeid snails in Vietnam. Identification of lymnaeid snails in Vietnam and their role in the liver fluke transmission should be further investigated.

Citations

Citations to this article as recorded by

Distribution and Fasciola infection rates of Lymnaea snails and cattle in high-salinity areas of Mekong Delta, Vietnam
Dang Thi LOAN, Lam Thanh NGUYEN, Tran Ngoc BICH, Nguyen Thuy Y VI, Yasunobu MATSUMOTO
Journal of Veterinary Medical Science.2025; 87(3): 291. CrossRef
Susceptibility of lymnaeid snails to Fasciola hepatica and Fasciola gigantica (Digenea: Fasciolidae): a systematic review and meta-analysis
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Na Li, Bin Hou, Wurilige Tian, Siyao Li, Buhe Buyin, Ying Hai, Surong Hasi
International Journal for Parasitology: Parasites and Wildlife.2024; 23: 100896. CrossRef
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Vinh Hoang Quang, Bruno Levecke, Dung Do Trung, Brecht Devleesschauwer, Binh Vu Thi Lam, Kathy Goossens, Katja Polman, Steven Callens, Pierre Dorny, Veronique Dermauw, Krystyna Cwiklinski
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Environmental influence on abundance and infection patterns of snail intermediate hosts of liver and intestinal flukes in North and Central Vietnam
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Parasitology Research.2024;[Epub] CrossRef
Assessment of the genetic diversity of lymnaeid (Gastropoda: Pulmonata) snails and their infection status with trematode cercariae in different regions of Thailand
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Infection, Genetics and Evolution.2024; 119: 105576. CrossRef
Mitochondrial genome of the fluke pond snail, Austropeplea cf. brazieri (Gastropoda: Lymnaeidae)
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Parasites & Vectors.2024;[Epub] CrossRef
A multidisciplinary analysis of over 53,000 fascioliasis patients along the 1995–2019 countrywide spread in Vietnam defines a new epidemiological baseline for One Health approaches
Nguyen Van De, Pham Ngoc Minh, Thanh Hoa Le, Do Trung Dung, Tran Thanh Duong, Bui Van Tuan, Le Thanh Dong, Nguyen Van Vinh Chau, Pablo F. Cuervo, M. Dolores Bargues, M. Adela Valero, Albis Francesco Gabrielli, Antonio Montresor, Santiago Mas-Coma
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Veterinary Parasitology: Regional Studies and Reports.2019; 17: 100318. CrossRef
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Mini Review

Paragonimus and Paragonimiasis in Vietnam: an Update: Pham Ngoc Doanh, Yoichiro Horii, Yukifumi Nawa; Korean J Parasitol 2013;51(6):621-627.
Published online December 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.6.621

Abstract
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Paragonimiasis is a food-borne parasitic zoonosis caused by infection with lung flukes of the genus Paragonimus. In Vietnam, research on Paragonimus and paragonimiasis has been conducted in northern and central regions of the country. Using a combination of morphological and molecular methods, 7 Paragonimus species, namely P. heterotremus, P. westermani, P. skrjabini, P. vietnamensis, P. proliferus, P. bangkokenis and P. harinasutai, have been identified in Vietnam. Of these, the first 3, P. heterotremus, P. westermani and P. skrjabini, are known to infect humans in other countries. However, in Vietnam, only P. heterotremus, found in some northern provinces, has been shown to infect humans. Even nowadays, local people in some northern provinces, such as Lai Chau and Yen Bai, are still suffering from P. heterotremus infection. In some provinces of central Vietnam, the prevalence and infection intensity of P. westermani metacercariae in freshwater crabs (the second intermediate hosts) are extremely high, but human cases have not been reported. Likewise, although P. skrjabini was found in Thanh Hoa Province, its pathogenicity to humans in Vietnam still remains uncertain. The results of molecular phylogenetic analyses of Vietnamese Paragonimus species provides new insights on the phylogeny and taxonomy of the genus Paragonimus. Comprehensive molecular epidemiological and geobiological studies on the genus in Vietnam and adjacent countries are needed to clarify the biodiversity and public health significance of the lung flukes.

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Lung flukes of the genusParagonimus: ancient and re-emerging pathogens
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Molecular and morphological variation of Paragonimus westermani in Vietnam with records of new second intermediate crab hosts and a new locality in a northern province
N. PHAM DOANH, A. LUU TU, T. DUNG BUI, T. HO LOAN, NARIAKI NONAKA, YOICHIRO HORII, DAVID BLAIR, YUKIFUMI NAWA
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