Parasites, Hosts and Diseases

"real-time PCR"

Original Article

Molecular cloning, identification, transcriptional analysis, and silencing of enolase on the life cycle of Haemaphysalis longicornis (Acari, Ixodidae) tick: Md. Samiul Haque, Md. Khalesur Rahman, Mohammad Saiful Islam, Myung-Jo You; Parasites Hosts Dis 2024;62(2):226-237.
Published online May 27, 2024; DOI: https://doi.org/10.3347/PHD.24015

Ticks, blood-sucking ectoparasites, spread diseases to humans and animals. Haemaphysalis longicornis is a significant vector for tick-borne diseases in medical and veterinary contexts. Identifying protective antigens in H. longicornis for an anti-tick vaccine is a key tick control strategy. Enolase, a multifunctional protein, significantly converts D-2-phosphoglycerate and phosphoenolpyruvate in glycolysis and gluconeogenesis in cell cytoplasm. This study cloned a complete open reading frame (ORF) of enolase from the H. longicornis tick and characterized its transcriptional and silencing effect. We amplified the full-length cDNA of the enolase gene using rapid amplification of cDNA ends. The complete cDNA, with an ORF of 1,297 nucleotides, encoded a 432-amino acid polypeptide. Enolase of the Jeju strain H. longicornis exhibited the highest sequence similarity with H. flava (98%), followed by Dermacentor silvarum (82%). The enolase motifs identified included N-terminal and C-terminal regions, magnesium binding sites, and several phosphorylation sites. Reverse transcription-polymerase chain reaction (RT-PCR) analysis indicated that enolase mRNA transcripts were expressed across all developmental stages of ticks and organs such as salivary gland and midgut. RT-PCR showed higher transcript levels in syn-ganglia, suggesting that synganglion nerves influence enolase,s role in tick salivary glands. We injected enolase double-stranded RNA into adult unfed female ticks, after which they were subsequently fed with normal unfed males until they spontaneously dropped off. RNA interference significantly (P<0.05) reduced feeding and reproduction, along with abnormalities in eggs (no embryos) and hatching. These findings suggest enolase is a promising target for future tick control strategies.

Citations

Citations to this article as recorded by

Comprehensive antigen identification and comparative analysis: significant approaches for controlling Haemaphysalis longicornis ticks
Md. Samiul Haque, Bumseok Kim, Myung-Jo You
Journal of Veterinary Science.2025;[Epub] CrossRef
Microbiome Composition of Haemaphysalis flava in Korea and Diversity Analysis Based on Region, Developmental Stage, and Sex
Min Kyu Sang, Jie eun Park, Dae Kwon Song, Jun Yang Jeong, Chan‐Eui Hong, Hyeonjun Shin, Hyeok Lee, Kyoung Won Lee, Hee Ju Hwang, Hyun woo Kim, Seong Yoon Kim, Wook‐Gyo Lee, So Young Park, Se Won Kang, Jung Han Park, Bharat Bhusan Patnaik, Sung‐Jae Cha, S
Entomological Research.2025;[Epub] CrossRef
Identification and knockdown effect of disulfide isomerase in the Haemaphysalis longicornis (Acari: Ixodidae)
Md Samiul Haque, Bumseok Kim, Myung-Jo You
Revista Brasileira de Parasitologia Veterinária.2025;[Epub] CrossRef

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Brief Communication

Clinical Usefulness of LabChip Real-time PCR using Lab-On-a-Chip Technology for Diagnosing Malaria: Jeeyong Kim, Da Hye Lim, Do-CiC Mihn, Jeonghun Nam, Woong Sik Jang, Chae Seung Lim; Korean J Parasitol 2021;59(1):77-82.
Published online February 19, 2021; DOI: https://doi.org/10.3347/kjp.2021.59.1.77

Abstract
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As malaria remains a major health problem worldwide, various diagnostic tests have been developed, including microscopy-based and rapid diagnostic tests. LabChip real-time PCR (LRP) is a small and portable device used to diagnose malaria using lab-on-a-chip technology. This study aimed to evaluate the diagnostic performance of LRP for detecting malaria parasites. Two hundred thirteen patients and 150 healthy individuals were enrolled from May 2009 to October 2015. A diagnostic detectability of LRP for malaria parasites was compared to that of conventional RT-PCR. Sensitivity of LRP for Plasmodium vivax, P. falciparum, P. malariae, and P. ovale was 95.5%, 96.0%, 100%, and 100%, respectively. Specificity of LRP for P. vivax, P. falciparum, P. malariae, and P. ovale was 100%, 99.3%, 100%, and 100%, respectively. Cohen’s Kappa coefficients between LRP and CFX96 for detecting P. vivax, P. falciparum, P. malariae, and P. ovale were 0.96, 0.98, 1.00, and 1.00, respectively. Significant difference was not observed between the results of LRP and conventional RT-PCR and microscopic examination. A time required to amplify DNAs using LRP and conventional RT-PCR was 27 min and 86 min, respectively. LRP amplified DNAs 2 times more fast than conventional RT-PCR due to the faster heat transfer. Therefore, LRP could be employed as a useful tool for detecting malaria parasites in clinical laboratories.

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Original Articles

Development of Molecular Diagnosis Using Multiplex Real-Time PCR and T4 Phage Internal Control to Simultaneously Detect Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis from Human Stool Samples: Ji-Hun Shin, Sang-Eun Lee, Tong Soo Kim, Da-Won Ma, Shin-Hyeong Cho, Jong-Yil Chai, Eun-Hee Shin; Korean J Parasitol 2018;56(5):419-427.
Published online October 31, 2018; DOI: https://doi.org/10.3347/kjp.2018.56.5.419

Abstract
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This study aimed to develop a new multiplex real-time PCR detection method for 3 species of waterborne protozoan parasites (Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis) identified as major causes of traveler’s diarrhea. Three target genes were specifically and simultaneously detected by the TaqMan probe method for multiple parasitic infection cases, including Cryptosporidium oocyst wall protein for C. parvum, glutamate dehydrogenase for G. lamblia, and internal transcribed spacer 1 for C. cayetanensis. Gene product 21 for bacteriophage T4 was used as an internal control DNA target for monitoring human stool DNA amplification. TaqMan probes were prepared using 4 fluorescent dyes, FAMTM, HEXTM, Cy5TM, and CAL Fluor Red® 610 on C. parvum, G. lamblia, C. cayetanensis, and bacteriophage T4, respectively. We developed a novel primer-probe set for each parasite, a primer-probe cocktail (a mixture of primers and probes for the parasites and the internal control) for multiplex real-time PCR analysis, and a protocol for this detection method. Multiplex real-time PCR with the primer-probe cocktail successfully and specifically detected the target genes of C. parvum, G. lamblia, and C. cayetanensis in the mixed spiked human stool sample. The limit of detection for our assay was 2×10 copies for C. parvum and for C. cayetanensis, while it was 2×103 copies for G. lamblia. We propose that the multiplex real-time PCR detection method developed here is a useful method for simultaneously diagnosing the most common causative protozoa in traveler’s diarrhea.

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Comparison of Three Real-Time PCR Assays Targeting the SSU rRNA Gene, the COWP Gene and the DnaJ-Like Protein Gene for the Diagnosis of Cryptosporidium spp. in Stool Samples
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Molecular Method Confirms Canine Leishmania Infection Detected by Serological Methods in Non-Endemic Area of Brazil: Emeline Riboldi, Flavio Carvalho, Pedro Roosevelt Torres Roma?o, Regina Bones Barcellos, Graziele Lima Bello, Raquel Rocha Ramos, Rosemari Terezinha de Oliveira, Joa?o Pessoa Arau?jo Ju?nior, Maria Lucia Rossetti, Eliane Dallegrave; Korean J Parasitol 2018;56(1):11-19.
Published online February 28, 2018; DOI: https://doi.org/10.3347/kjp.2018.56.1.11

Abstract
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In Brazil, visceral leishmaniasis (VL) is expanding and becoming urbanized, especially in non-endemic areas such as the State of Rio Grande do Sul. Considering that infected dogs are the main reservoir for zoonotic VL, this study evaluated the prevalence of canine visceral leishmaniasis (CVL) in the metropolitan area of Porto Alegre, a new area of ex- pansion of VL in Brazil. Serum and plasma from 405 asymptomatic dogs from the municipalities of Canoas (n = 107), Sa?o Leopoldo (n=216), and Novo Hamburgo (n=82) were tested for CVL using immunochromatographic (DPP^®) and ELISA EIE^® assays (2 assays officially adopted by the Brazilian government for the diagnosis of CVL) and real-time PCR to con- firm the results. There was no agreement among serological and real-time PCR results, indicating that the Leishmania infection in asymptomatic animals with low parasite load, confirmed by negative parasitological tests (smears and parasite culture), need to be evaluated by molecular methods. The prevalence of LVC in the metropolitan region of Porto Alegre, confirmed by real-time PCR was 4% (5.6% in Canoas and 4.6% in Sa?o Leopoldo). The use of molecular method is essential for accurate diagnosis of CVL, especially in asymptomatic dogs in non-endemic areas.

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Development and preliminary study of the rLiNTPDase2 rapid test: A lateral flow immunochromatographic assay for Canine Visceral Leishmaniasis
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Simultaneous Molecular Detection of Cryptosporidium and Cyclospora from Raw Vegetables in Korea: Seobo Sim, Jua Won, Jae-Whan Kim, Kyungjin Kim, Woo-Yoon Park, Jae-Ran Yu; Korean J Parasitol 2017;55(2):137-142.
Published online April 30, 2017; DOI: https://doi.org/10.3347/kjp.2017.55.2.137

Abstract
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Cryptosporidium and Cyclospora are well-known coccidian protozoa that can cause waterborne and foodborne diarrheal illnesses. There have been a few reports regarding contamination in different vegetables with Cryptosporidium, but no data are available regarding the sources of Cyclospora infections in Korea. In the present study, we collected 6 kinds of vegetables (perilla leaves, winter-grown cabbages, chives, sprouts, blueberries, and cherry tomatoes) from July 2014 to June 2015, and investigated contamination by these 2 protozoa using multiplex quantitative real-time PCR. Among 404 vegetables, Cryptosporidium and Cyclospora were detected in 31 (7.7%) and 5 (1.2%) samples, respectively. In addition, Cryptosporidium was isolated from all 6 kinds of vegetables, whereas Cyclospora was detected in 4 kinds of vegetables (except perilla leaves and chives). Cryptosporidium (17.8%) and Cyclospora (2.9%) had the highest detection rates in chives and winter-grown cabbages, respectively. Cryptosporidium was detected all year long; however, Cyclospora was detected only from October to January. In 2 samples (sprout and blueberry), both Cryptosporidium and Cyclospora were detected. Further investigations using TaqI restriction enzyme fragmentation and nested PCR confirmed Cryptosporidium parvum and Cyclospora cayetanensis, respectively. In conclusion, we detected C. cayetanensis in vegetables for the first time in Korea. This suggests that screening should be employed to prevent these protozoal infections in Korea.

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Assessment of pesticide residues and microbial contamination in raw leafy green vegetables marketed in Italy
Gino Angelo Santarelli, Giacomo Migliorati, Francesco Pomilio, Cristina Marfoglia, Patrizia Centorame, Antonella D'Agostino, Roberta D'Aurelio, Rossana Scarpone, Noemi Battistelli, Federica Di Simone, Giuseppe Aprea, Luigi Iannetti
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Differential Expression of Hox and Notch Genes in Larval and Adult Stages of Echinococcus granulosus: Ebrahim Saedi Dezaki, Mohammad Mehdi Yaghoobi, Elham Taheri, Pooya Ghaseminejad Almani, Farideh Tohidi, Bruno Gottstein, Majid Fasihi Harandi; Korean J Parasitol 2016;54(5):653-658.
Published online October 31, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.5.653

Abstract
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This investigation aimed to evaluate the differential expression of HoxB7 and notch genes in different developmental stages of Echinococcus granulosus sensu stricto. The expression of HoxB7 gene was observed at all developmental stages. Nevertheless, significant fold differences in the expression level was documented in the juvenile worm with 3 or more proglottids, the germinal layer from infected sheep, and the adult worm from an experimentally infected dog. The notch gene was expressed at all developmental stages of E. granulosus; however, the fold difference was significantly increased at the microcysts in monophasic culture medium and the germinal layer of infected sheep in comparison with other stages. The findings demonstrated that the 2 aforementioned genes evaluated in the present study were differentially expressed at different developmental stages of the parasite and may contribute to some important biological processes of E. granulosus.

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Yan Lv, Xuan Pang, Zhonghong Cao, Changping Song, Baohua Liu, Weiwei Wu, Qiuxiang Pang
International Journal of Molecular Sciences.2024; 25(6): 3322. CrossRef
Echinococcus multilocularis delta/notch signalling components are expressed in post-mitotic cells
Chris Speicher, Monika Bergmann, Klaus Brehm
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Notch signaling pathway involved in Echinococcus granulosus infection regulates dendritic cell development and differentiation
Mingxia Wang, Zailing Shang, Fei Qiao, Junhu Hei, Xueling Ma, Yana Wang
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Proteomic profiling of hydatid fluid from pulmonary cystic echinococcosis
Guilherme Brzoskowski dos Santos, Edileuza Danieli da Silva, Eduardo Shigueo Kitano, Maria Eduarda Battistella, Karina Mariante Monteiro, Jeferson Camargo de Lima, Henrique Bunselmeyer Ferreira, Solange Maria de Toledo Serrano, Arnaldo Zaha
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The Genomes of Two Strains of Taenia crassiceps the Animal Model for the Study of Human Cysticercosis
Raúl J. Bobes, Karel Estrada, Diana G. Rios-Valencia, Arturo Calderón-Gallegos, Patricia de la Torre, Julio C. Carrero, Alejandro Sanchez-Flores, Juan P. Laclette
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Bioinformatics analysis and experimental verification of Notch signalling pathway-related miRNA–mRNA subnetwork in extracellular vesicles during Echinococcus granulosus encystation
Jian Gao, Xuan Zhou, Ling Liu, Guodong Lv, Qiulian Hou, Xiaofan Zhang, Yujuan Shen
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Screening, construction, and serological identification of the diagnostic antigen molecule EG-06283 for the diagnosis of Echinococcus granulosus
Yazhou Zhu, Tingrui Zhang, Yinqi Zhao, Yin Wang, Yongxue Lv, Shasha Li, Songhao Yang, Mingxing Zhu, Wei Zhao, Qiang Wang
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Evolutionary Transformations of the Metazoan Body Plan: Genomic-Morphogenetic Correlations
V. V. Isaeva, S. V. Rozhnov
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In vitro effects of glutathione on Transforming Growth Factor beta and Epidermal Growth Factor Receptor genes expression in the protoscoleces and strobilated worms of Echinococcus granulosus
Seyyed Jafar Nosratabadi, Nasim Hayati Roodbari, Ali Afgar, Mohammad Hossein Modarressi, Alireza Farsinejad, Majid Fasihi Harandi
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T. Basika, G.P. Paludo, F.M. Araujo, A.C. Salim, F. Pais, L. Maldonado, N. Macchiaroli, J. Camargo de Lima, M. Rosenzvit, G.C. Oliveira, L. Kamenetzky, H.B. Ferreira
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Calmodulin-specific small interfering RNA induces consistent expression suppression and morphological changes in Echinococcus granulosus
Seyed Mohammad Mousavi, Ali Afgar, Mohammad Ali Mohammadi, Seifollah Mortezaei, Balal Sadeghi, Majid Fasihi Harandi
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Validation of Reference Genes for Quantitative Real-Time PCR in Bovine PBMCs Transformed and Non-transformed by Theileria annulata: Hongxi Zhao, Junlong Liu, Youquan Li, Congshan Yang, Shuaiyang Zhao, Juan Liu, Aihong Liu, Guangyuan Liu, Hong Yin, Guiquan Guan, Jianxun Luo; Korean J Parasitol 2016;54(1):39-46.
Published online February 26, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.1.39

Abstract
PDF

Theileria annulata is a tick-borne intracellular protozoan parasite that causes tropical theileriosis, a fatal bovine lymphoproliferative disease. The parasite predominantly invades bovine B lymphocytes and macrophages and induces host cell transformation by a mechanism that is not fully comprehended. Analysis of signaling pathways by quantitative real-time PCR (qPCR) could be a highly efficient means to understand this transformation mechanism. However, accurate analysis of qPCR data relies on selection of appropriate reference genes for normalization, yet few papers on T. annulata contain evidence of reference gene validation. We therefore used the geNorm and NormFinder programs to evaluate the stability of 5 candidate reference genes; 18S rRNA, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ACTB (β-actin), PRKG1 (protein kinase cGMP-dependent, type I) and TATA box binding protein (TBP). The results showed that 18S rRNA was the reference gene most stably expressed in bovine PBMCs transformed and non-transformed with T. annulata, followed by GAPDH and TBP. While 18S rRNA and GAPDH were the best combination, these 2 genes were chosen as references to study signaling pathways involved in the transformation mechanism of T. annulata.

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Elhem Yacoub, Daniel Kos, Murray Jelinski
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Shahin Tajeri, Debasish Chattopadhyay, Gordon Langsley, Ard M. Nijhof, Vikrant Sudan
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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
PDF

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.

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Paola Cociancic, Jacklyn Comas, J. Guillermo Esteban
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Narut Butploy, Wanida Kanarkard, Pewpan M. Intapan, Oranuch Sanpool
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Fish and Food-Fatale: Food-borne Trematode Opisthorchis viverrini and Cholangiocarcinoma
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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
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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
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Xialei Liu, Genglong Zhu, Chaonong Cai, Zhiyue Lv, Jian Li
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Vivornpun Sanprasert, Ruthairat Kerdkaew, Siriporn Srirungruang, Sarit Charuchaibovorn, Kobpat Phadungsaksawasdi, Surang Nuchprayoon
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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
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Molecular Differentiation of Schistosoma japonicum and Schistosoma mekongi by Real-Time PCR with High Resolution Melting Analysis: Amornmas Kongklieng, Worasak Kaewkong, Pewpan M. Intapan, Oranuch Sanpool, Penchom Janwan, Tongjit Thanchomnang, Viraphong Lulitanond, Pusadee Sri-Aroon, Yanin Limpanont, Wanchai Maleewong; Korean J Parasitol 2013;51(6):651-656.
Published online December 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.6.651

Abstract
PDF

Human schistosomiasis caused by Schistosoma japonicum and Schistosoma mekongi is a chronic and debilitating helminthic disease still prevalent in several countries of Asia. Due to morphological similarities of cercariae and eggs of these 2 species, microscopic differentiation is difficult. High resolution melting (HRM) real-time PCR is developed as an alternative tool for the detection and differentiation of these 2 species. A primer pair was designed for targeting the 18S ribosomal RNA gene to generate PCR products of 156 base pairs for both species. The melting points of S. japonicum and S. mekongi PCR products were 84.5±0.07℃ and 85.7±0.07℃, respectively. The method permits amplification from a single cercaria or an egg. The HRM real-time PCR is a rapid and simple tool for differentiation of S. japonicum and S. mekongi in the intermediate and final hosts.

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Adrian Miki C. Macalanda, Atcharaphan Wanlop, Kevin Austin L. Ona, Eloiza May S. Galon, Virak Khieu, Somphou Sayasone, Aya Yajima, Jose Ma. M. Angeles, Shin-ichiro Kawazu
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Chao Lv, Wangping Deng, Liping Wang, Zhiqiang Qin, Xiaonong Zhou, Jing Xu
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Rapid Detection and Identification of Wuchereria bancrofti, Brugia malayi, B. pahangi, and Dirofilaria immitis in Mosquito Vectors and Blood Samples by High Resolution Melting Real-Time P: Tongjit Thanchomnang, Pewpan M. Intapan, Chairat Tantrawatpan, Viraphong Lulitanond, Sudchit Chungpivat, Piyanan Taweethavonsawat, Worasak Kaewkong, Oranuch Sanpool, Penchom Janwan, Wej Choochote, Wanchai Maleewong; Korean J Parasitol 2013;51(6):645-650.
Published online December 31, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.6.645

Abstract
PDF

A simple, rapid, and high-throughput method for detection and identification of Wuchereria bancrofti, Brugia malayi, Brugia pahangi, and Dirofilaria immitis in mosquito vectors and blood samples was developed using a real-time PCR combined with high-resolution melting (HRM) analysis. Amplicons of the 4 filarial species were generated from 5S rRNA and spliced leader sequences by the real-time PCR and their melting temperatures were determined by the HRM method. Melting of amplicons from W. bancrofti, B. malayi, D. immitis, and B. pahangi peaked at 81.5±0.2℃, 79.0±0.3℃, 76.8±0.1℃, and 79.9±0.1℃, respectively. This assay is relatively cheap since it does not require synthesis of hybridization probes. Its sensitivity and specificity were 100%. It is a rapid and technically simple approach, and an important tool for population surveys as well as molecular xenomonitoring of parasites in vectors.

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Brief Communication

Cryptosporidium hominis Infection Diagnosed by Real-Time PCR-RFLP: Hyeng-Il Cheun, Kyungjin Kim, Sejoung Yoon, Won-Ja Lee, Woo-Yoon Park, Seobo Sim, Jae-Ran Yu; Korean J Parasitol 2013;51(3):353-355.
Published online June 30, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.3.353

Abstract
PDF

There are approximately 20 known species of the genus Cryptosporidium, and among these, 8 infect immunocompetent or immunocompromised humans. C. hominis and C. parvum most commonly infect humans. Differentiating between them is important for evaluating potential sources of infection. We report here the development of a simple and accurate real-time PCR-based restriction fragment length polymorphism (RFLP) method to distinguish between C. parvum and C. hominis. Using the CP2 gene as the target, we found that both Cryptosporidium species yielded 224 bp products. In the subsequent RFLP method using TaqI, 2 bands (99 and 125 bp) specific to C. hominis were detected. Using this method, we detected C. hominis infection in 1 of 21 patients with diarrhea, suggesting that this method could facilitate the detection of C. hominis infections.

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Case Report

A Case of Plasmodium ovale Malaria Imported from West Africa: Yunjung Kang, Jinyoung Yang; Korean J Parasitol 2013;51(2):213-218.
Published online April 25, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.2.213

Abstract
PDF

Malaria is a parasitic infection caused by Plasmodium species. Most of the imported malaria in Korea are due to Plasmodium vivax and Plasmodium falciparum, and Plasmodium ovale infections are very rare. Here, we report a case of a 24-year-old American woman who acquired P. ovale while staying in Ghana, West Africa for 5 months in 2010. The patient was diagnosed with P. ovale malaria based on a Wright-Giemsa stained peripheral blood smear, Plasmodium genus-specific real-time PCR, Plasmodium species-specific nested PCR, and sequencing targeting 18S rRNA gene. The strain identified had a very long incubation period of 19-24 months. Blood donors who have malaria with a very long incubation period could be a potential danger for propagating malaria. Therefore, we should identify imported P. ovale infections not only by morphological findings but also by molecular methods for preventing propagation and appropriate treatment.

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Original Article

Detection of Plasmodium vivax by Nested PCR and Real-Time PCR: Genc, Ahmet , Eroglu, Fadime , Koltas, Ismail Soner; Korean J Parasitol 2010;48(2):99-103.; DOI: https://doi.org/10.3347/kjp.2010.48.2.99

Abstract
PDF

Malaria is endemic in the Cukurova region while it is sporadic in other regions of Turkey. Therefore, the laboratory and clinical diagnosis of malaria is important for the treatment of malaria. In this study, 92 blood samples that were taken from the suspected malaria patients for routine diagnosis in a period of 10 years between 1999 and 2009 were analyzed. All of these blood samples were examined by microscopic examinations using Giemsa-stained thick blood films, nested PCR, and real-time PCR. The sensitivity-specificity and positive-negative predictive values for these diagnostic tests were then calculated. It was found that the positive predictive values of microscopic examination of thick blood films, nested PCR, and real-time PCR were 47.8%, 56.5%, and 60.9% for malaria, respectively. The real-time PCR was found to have a specificity of 75% and sensitivity of 100%, while specificity and sensitivity of nested PCR was found 81.2% and 97.7% according to the microscopic examination of thick blood films, respectively.

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