Parasites, Hosts and Diseases

"multiplex PCR"

Brief Communication

Comparative Assessment of Diagnostic Performance of Cytochrome Oxidase Multiplex PCR and 18S rRNA Nested PCR: Preeti Kumari, Swati Sinha, Renuka Gahtori, Afshana Quadiri, Paras Mahale, Deepali Savargaonkar, Veena Pande, Bina Srivastava, Himmat Singh, Anupkumar R Anvikar; Korean J Parasitol 2022;60(4):295-299.
Published online August 24, 2022; DOI: https://doi.org/10.3347/kjp.2022.60.4.295

Malaria elimination and control require prompt and accurate diagnosis for treatment plan. Since microscopy and rapid diagnostic test (RDT) are not sensitive particularly for diagnosing low parasitemia, highly sensitive diagnostic tools are required for accurate treatment. Molecular diagnosis of malaria is commonly carried out by nested polymerase chain reaction (PCR) targeting 18S rRNA gene, while this technique involves long turnaround time and multiple steps leading to false positive results. To overcome these drawbacks, we compared highly sensitive cytochrome oxidase gene-based single-step multiplex reaction with 18S rRNA nested PCR. Cytochrome oxidase (cox) genes of P. falciparum (cox-III) and P. vivax (cox-I) were compared with 18S rRNA gene nested PCR and microscopy. Cox gene multiplex PCR was found to be highly specific and sensitive, enhancing the detection limit of mixed infections. Cox gene multiplex PCR showed a sensitivity of 100% and a specificity of 97%. This approach can be used as an alternative diagnostic method as it offers higher diagnostic performance and is amenable to high throughput scaling up for a larger sample size at low cost.

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Therapeutic mucosal vaccination of herpes simplex virus type 2 infected guinea pigs with an adenovirus-based vaccine expressing the ribonucleotide reductase 2 and glycoprotein D induces local tissue-resident CD4+ and CD8+ TRM cells associated with protect
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Validation of real-time PCR assays for detecting Plasmodium and Babesia DNA species in blood samples
Luz Helena Patiño, Sergio Castañeda, Milena Camargo, Li Yong Cao, Bernadette Liggayu, Alberto Paniz‐Mondolfi, Juan David Ramírez
Acta Tropica.2024; 258: 107350. CrossRef
Comparative Analysis of Multiplex/Semi-nested PCR and Microscopy for Accurate Human Malaria Species Diagnosis
Aram Khezri, Mehdi Nateghpour, Haleh Hanifian, Leila Farivar, Afsaneh Motevalli Haghi
Jundishapur Journal of Microbiology.2024;[Epub] CrossRef

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Original Article

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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Chenhang Jiang, Yang Li, Ping Yu, Mengjun Fang, Di Huang, Xiangming Fang, Zhinan Xu
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Lynn Glyschewski, Hagen Frickmann, Fred Stephen Sarfo, Betty Roberta Norman, Albert Dompreh, Emmanuel Acheamfour-Akowuah, Martin Kofi Agyei, Shadrack Osei Asibey, Richard Boateng, Edmund Osei Kuffour, Veronica Di Cristanziano, Sven Poppert, Felix Weinreic
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Oumaima Hachimi, Rebecca Falender, Gabriel Davis, Rispa Vranka Wafula, Melissa Sutton, June Bancroft, Paul Cieslak, Christine Kelly, Devrim Kaya, Tyler Radniecki
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Atul Kumar Sharma, K. Gururaj, Rama Sharma, Anjana Goel, Souvik Paul, Dinesh Kumar Sharma
Cell Biochemistry and Function.2023; 41(1): 45. CrossRef
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Stefania Nicola, Francesco Cinetto, Stefano Della Mura, Luca Lo Sardo, Elena Saracco, Ilaria Vitali, Riccardo Scarpa, Helena Buso, Vera Bonato, Claudia Discardi, Giovanni Rolla, Carla Felice, Marcello Rattazzi, Luisa Brussino
Biomedicines.2023; 11(1): 170. CrossRef
Efficacy of a membrane concentration method combined with real-time PCR for detection of Giardia and Cryptosporidium in drinking water
Jiang Jingyi, Yao Ping, Xu Jian, Chen Jia, Mao Xujian, Li Qiong, Tu Bowen, Wang Fengming
Letters in Applied Microbiology.2023;[Epub] CrossRef
Bacteriophages: from Isolation to Application
Abdallah Abdelsattar, Alyaa Dawoud, Salsabil Makky , Rana Nofal, Ramy Aziz, Ayman El-Shibiny
Current Pharmaceutical Biotechnology.2022; 23(3): 337. CrossRef
Comparative Evaluation of Real-Time Screening PCR Assays for Giardia duodenalis and of Assays Discriminating the Assemblages A and B
Felix Weinreich, Andreas Hahn, Kirsten Alexandra Eberhardt, Simone Kann, Torsten Feldt, Fred Stephen Sarfo, Veronica Di Cristanziano, Hagen Frickmann, Ulrike Loderstädt
Microorganisms.2022; 10(7): 1310. CrossRef
Review on Cyclosporiasis Outbreaks and Potential Molecular Markers for Tracing Back Investigations
Junqiang Li, Feifei Xu, Md Robiul Karim, Longxian Zhang
Foodborne Pathogens and Disease.2022; 19(12): 796. CrossRef
Multiplex Molecular Point-of-Care Test for Syndromic Infectious Diseases
Hanbi Kim, Hee Jae Huh, Eunkyoung Park, Doo-Ryeon Chung, Minhee Kang
BioChip Journal.2021; 15(1): 14. CrossRef
Comparative Performance of Eight PCR Methods to Detect Cryptosporidium Species
Damien Costa, Louise Soulieux, Romy Razakandrainibe, Louise Basmaciyan, Gilles Gargala, Stéphane Valot, Frédéric Dalle, Loic Favennec
Pathogens.2021; 10(6): 647. CrossRef
A review on application of next-generation sequencing methods for profiling of protozoan parasites in water: Current methodologies, challenges, and perspectives
N.P. Mthethwa, I.D. Amoah, P. Reddy, F. Bux, S. Kumari
Journal of Microbiological Methods.2021; 187: 106269. CrossRef
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
Felix Weinreich, Andreas Hahn, Kirsten Alexandra Eberhardt, Torsten Feldt, Fred Stephen Sarfo, Veronica Di Cristanziano, Hagen Frickmann, Ulrike Loderstädt
Pathogens.2021; 10(9): 1131. CrossRef
Advances in Cyclosporiasis Diagnosis and Therapeutic Intervention
Junqiang Li, Zhaohui Cui, Meng Qi, Longxian Zhang
Frontiers in Cellular and Infection Microbiology.2020;[Epub] CrossRef
Comparison of commercial and in-house real-time PCR platforms for 15 parasites and microsporidia in human stool samples without a gold standard
Thomas Köller, Andreas Hahn, Enkhtsetseg Altangerel, Jaco J. Verweij, Olfert Landt, Simone Kann, Denise Dekker, Jürgen May, Ulrike Loderstädt, Andreas Podbielski, Hagen Frickmann
Acta Tropica.2020; 207: 105516. CrossRef
Biochemical Markers of the Functional State of the Liver during Giardiasis
D. V. Morozenko, K. V. Gliebova, S. V. Ivannikova, O. G. Geyderikh, O. V. Shapovalova, A. V. Derevyanko
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R.C.A. Thompson, A. Ash
Infection, Genetics and Evolution.2019; 75: 103951. CrossRef
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Sonia Almeria, Hediye N. Cinar, Jitender P. Dubey
Microorganisms.2019; 7(9): 317. CrossRef

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Brief Communication

Differential Diagnosis of Human Sparganosis Using Multiplex PCR: Hyeong-Kyu Jeon, Kyu-Heon Kim, Woon-Mok Sohn, Keeseon S. Eom; Korean J Parasitol 2018;56(3):295-300.
Published online June 30, 2018; DOI: https://doi.org/10.3347/kjp.2018.56.3.295

Abstract
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Human sparganosis was diagnosed by morphological and genetic analyses in Korea. The complete mitochondrial genomes of Spirometra erinaceieuropaei and S. decipiens isolated in Korea have been recorded. Present study was performed to provide information to diagnose the etiologic agent of sparganosis by multiplex PCR using mitochondrial genome sequences of S. erinaceieuropaei and S. decipiens. In an effort to examine the differential diagnosis of spirometrid tapeworms, multiplex PCR assays were performed on plerocercoid larvae of S. erinaceieuropaei and S. decipiens. The PCR products obtained using species-specific primers were positively detected in all PCR assays on mixture of S. erinaceieuropaei and S. decipiens DNA. S. erinaceieuropaei-specific bands (239 bp and 401 bp) were obtained from all PCR assays using a mixture of S. erinaceieuropaei-specific primers (Se/Sd-1800F and Se-2018R; Se/Sd-7955F and Se-8356R) and S. erinaceieuropaei template DNA. S. decipiens-specific bands (540 bp and 644 bp) were also detected in all PCR assays containing mixtures of S. decipiens-specific primers (Se/Sd-1800F and Sd-2317R; Se/Sd-7955F and Sd-8567R) and S. decipiens template DNA. Sequence analyses on these species-specific bands revealed 100% sequence identity with homologous regions of the mtDNA sequences of S. erinaceieuropaei and S. decipiens. The multiplex PCR assay was useful for differential diagnosis of human sparganosis by detecting different sizes in species-specific bands.

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Irene Alvarado-Hidalgo, Josué Campos-Camacho, Yuliana Arguedas-Morales, Luis M. Romero-Vega, Alejandro Alfaro-Alarcón, Gabriela Anchia-Ureña, Laura G. Bass, Ivan Berrocal-Ávila, Isabel Hagnauer, Roberto W.I. Olivares, Alberto Solano-Barquero, Rodolfo Trau
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Epidemiology, Diagnosis, and Prevention of Sparganosis in Asia
Wei Liu, Tengfang Gong, Shuyu Chen, Quan Liu, Haoying Zhou, Junlin He, Yong Wu, Fen Li, Yisong Liu
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Alexander Morales, Rebeca M. Laird-Pérez, Virginia Capó, Enrique Iglesias, Luis Fonte, Arturo Plascencia-Hernández, Enrique J. Calderón, Keeseon S. Eom, Yaxsier de Armas, Héctor R. Pérez-Gómez
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Blaine A. Mathison, Sarah G. H. Sapp
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Large-scale survey of a neglected agent of sparganosis Spirometra erinaceieuropaei (Cestoda: Diphyllobothriidae) in wild frogs in China
Xi Zhang, Xiu Hong, Shi Nan Liu, Peng Jiang, Shu Chuan Zhao, Chuan Xi Sun, Zhong Quan Wang, Jing Cui, David Blair
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Original Article

Multiplex-Touchdown PCR to Simultaneously Detect Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the Major Causes of Traveler’s Diarrhea: Ji-Hun Shin, Sang-Eun Lee, Tong Soo Kim, Da-Won Ma, Jong-Yil Chai, Eun-Hee Shin; Korean J Parasitol 2016;54(5):631-636.
Published online October 31, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.5.631

Abstract
PDF

This study aimed to develop a multiplex-touchdown PCR method to simultaneously detect 3 species of protozoan parasites, i.e., Cryptosporidium parvum, Giardia lamblia, and Cyclospora cayetanensis, the major causes of traveler’s diarrhea and are resistant to standard antimicrobial treatments. The target genes included the Cryptosporidium oocyst wall protein for C. parvum, Glutamate dehydrogenase for G. lamblia, and 18S ribosomal RNA (18S rRNA) for C. cayetanensis. The sizes of the amplified fragments were 555, 188, and 400 bps, respectively. The multiplex-touchdown PCR protocol using a primer mixture simultaneously detected protozoa in human stools, and the amplified gene was detected in >1×10³ oocysts for C. parvum, >1×10⁴ cysts for G. lamblia, and >1 copy of the 18S rRNA gene for C. cayetanensis. Taken together, our protocol convincingly demonstrated the ability to simultaneously detect C. parvum, G. lamblia, and C. cayetanenesis in stool samples.

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A Multiplex RT-PCR for the Detection of Three Viruses and One Viroid Infecting Hemp
Derrick J. Grunwald, Jacob MacWilliams, Laine Hackenberg, Sydney Stroschein, Renee Rioux, Punya Nachappa, Shelby Ellison
PhytoFrontiers™.2024; 4(4): 524. CrossRef
Efficacy of a membrane concentration method combined with real-time PCR for detection of Giardia and Cryptosporidium in drinking water
Jiang Jingyi, Yao Ping, Xu Jian, Chen Jia, Mao Xujian, Li Qiong, Tu Bowen, Wang Fengming
Letters in Applied Microbiology.2023;[Epub] CrossRef
Screening of specific nucleic acid targets for Cronobacter sakazakii and visual detection by loop-mediated isothermal amplification and lateral flow dipstick method in powdered infant formula
Shiqian Fu, Xue Qin, Zhenghui Wang, Xinyan Yang, Sihan Chen, Tao Yang, Haonan Jin, Chaoxin Man, Yujun Jiang
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Luis Enrique Jerez Puebla, Fidel A. Núñez Fernández, Jorge Fraga Nodarse, Iraís Atencio Millán, Iredys Cruz Rodríguez, Lisette Pérez Santos, Yanet Pintos Saavedra, Laura Rodríguez Moreno, Lucy J. Robertson
Diagnostic Microbiology and Infectious Disease.2020; 97(3): 115048. CrossRef
Human cyclosporiasis
Annunziata Giangaspero, Robin B Gasser
The Lancet Infectious Diseases.2019; 19(7): e226. CrossRef
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Sonia Almeria, Hediye N. Cinar, Jitender P. Dubey
Microorganisms.2019; 7(9): 317. CrossRef
Evaluation of two DNA extraction methods for the PCR-based detection of eukaryotic enteric pathogens in fecal samples
Estelle Menu, Charles Mary, Isabelle Toga, Didier Raoult, Stéphane Ranque, Fadi Bittar
BMC Research Notes.2018;[Epub] CrossRef
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
The Korean Journal of Parasitology.2018; 56(5): 419. CrossRef
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Brief Communication

Loop-Mediated Isothermal Amplification Targeting Actin DNA of Trichomonas vaginalis: Youn-Kyoung Goo, Won-Sik Shin, Hye-Won Yang, So-Young Joo, Su-Min Song, Jae-Sook Ryu, Hyun-Hee Kong, Won-Ki Lee, Dong-Il Chung, Yeonchul Hong; Korean J Parasitol 2016;54(3):329-334.
Published online June 30, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.3.329

Abstract
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Trichomoniasis caused by Trichomonas vaginalis is a common sexually transmitted disease. Its association with several health problems, including preterm birth, pelvic inflammatory disease, cervical cancer, and transmission of human immunodeficiency virus, emphasizes the importance of improved access to early and accurate detection of T. vaginalis. In this study, a rapid and efficient loop-mediated isothermal amplification-based method for the detection of T. vaginalis was developed and validated, using vaginal swab specimens from subjects suspected to have trichomoniasis. The LAMP assay targeting the actin gene was highly sensitive with detection limits of 1 trichomonad and 1 pg of T. vaginalis DNA per reaction, and specifically amplified the target gene only from T. vaginalis. Validation of this assay showed that it had the highest sensitivity and better agreement with PCR (used as the gold standard) compared to microscopy and multiplex PCR. This study showed that the LAMP assay, targeting the actin gene, could be used to diagnose early infections of T. vaginalis. Thus, we have provided an alternative molecular diagnostic tool and a point-of-care test that may help to prevent trichomoniasis transmission and associated complications.

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Loop‐Mediated Isothermal Amplification (LAMP) for the Diagnosis of Sexually Transmitted Infections: A Review
Yasaman Ahmadi, Yejiong Yu, Zhanfeng Cui, Wei E. Huang, Monique I. Andersson
Microbial Biotechnology.2025;[Epub] CrossRef
A novel detection method based on MIRA-CRISPR/Cas13a-LFD targeting the repeated DNA sequence of Trichomonas vaginalis
Zhenke Yang, Jinghui Wang, Yiming Qi, Yiping Shi, Fakun Li, Weijuan Wang, Xiaowei Tian, Xuefang Mei, Zhenchao Zhang, Shuai Wang
Parasites & Vectors.2024;[Epub] CrossRef
A novel fluoro colorimetric Loop mediated isothermal amplification (LAMP) assay for detection of Trichomonas vaginalis
Shoorashetty Manohar Rudresh, Pareyam Pooja, Pattacheravanda Nanaiah Shakuntala, Kanta Madhu
Indian Journal of Medical Microbiology.2024; 49: 100610. CrossRef
Establishment of a programmatic detection method for Trichomonas vaginalis based on double antibody sandwich ELISA targeting TvCP39 antigen
Yuhua Li, Fakun Li, Wenjie Tian, Yani Zhang, Weijuan Wang, Zhenke Yang, Xiaowei Tian, Shuai Wang, Xuefang Mei, Zhenchao Zhang
Acta Tropica.2024; 260: 107489. CrossRef
Label-free electrochemical DNA biosensing of MR TV 29 18s ribosomal RNA gene of Trichomonas vaginalis by signalization of non-spherical gold nanoparticles
R. Dehdari Vais, H. Heli, N. Sattarahmady
Materials Today Communications.2023; 34: 105123. CrossRef
Construction a novel detection method for Trichomonas vaginalis based on recombinant enzyme polymerase amplification targeting the Actin gene
Fakun Li, Yangyang Deng, Wanxin Sheng, Xihui Gao, Weijuan Wang, Zhili Chu, Xuefang Mei, Zhenke Yang, Xiaowei Tian, Shuai Wang, Zhenchao Zhang
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Sebastián Lorenzo-Benito, Luis Alberto Rivera-Rivas, Lizbeth Sánchez-Ayala, Jaime Ortega-López, Octavio Montes-Flores, Daniel Talamás-Lara, Rossana Arroyo
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S. Ilbeigi, R. Dehdari Vais, N. Sattarahmady
Photodiagnosis and Photodynamic Therapy.2021; 34: 102290. CrossRef
Loop mediated isothermal amplification assay for detection of Trichomonas vaginalis in vaginal swabs among symptomatic women from North India
S. Khurana, R. Dadwal, N. Sharma, A. Mewara, S. Singh, R. Bagga, R. Yadav, S. Sethi
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Junjun Zhai, Zhang Yan, Feng Ping, Qu Lei, Xuelong Chen, Yanping Qi, Tianwen Wang
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Yuhua Li, Shuai Wang, Haoran Li, Xiaoxiao Song, Hao Zhang, Yujuan Duan, Chengyang Luo, Bingli Wang, Sifan Ji, Qing Xie, Zhenchao Zhang
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Label-free ultrasensitive electrochemical genosensing of Trichomonas vaginalis using anisotropic-shaped gold nanoparticles as a platform, a repeated sequence of the parasite DNA as a probe, and toluidine blue as a redox marker
N. Delshadi-Jahromi, R. Nazari-Vanani, H. Yadegari, N. Sattarahmady, G.R. Hatam, H. Heli
Sensors and Actuators B: Chemical.2018; 273: 234. CrossRef
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Original Article

Finding Wolbachia in Filarial larvae and Culicidae Mosquitoes in Upper Egypt Governorate: Ahmed K. Dyab, Lamia A. Galal, Abeer E. Mahmoud, Yasser Mokhtar; Korean J Parasitol 2016;54(3):265-272.
Published online June 30, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.3.265

Abstract
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Wolbachia is an obligatory intracellular endosymbiotic bacterium, present in over 20% of all insects altering insect reproductive capabilities and in a wide range of filarial worms which is essential for worm survival and reproduction. In Egypt, no available data were found about Wolbachia searching for it in either mosquitoes or filarial worms. Thus, we aimed to identify the possible concurrent presence of Wolbachia within different mosquitoes and filarial parasites, in Assiut Governorate, Egypt using multiplex PCR. Initially, 6 pools were detected positive for Wolbachia by single PCR. The simultaneous detection of Wolbachia and filarial parasites (Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens) by multiplex PCR was spotted in 5 out of 6 pools, with an overall estimated rate of infection (ERI) of 0.24%. Unexpectedly, the highest ERI (0.53%) was for Anopheles pharoensis with related Wolbachia and W. bancrofti, followed by Aedes (0.42%) and Culex (0.26%). We also observed that Wolbachia altered Culex spp. as a primary vector for W. bancrofti to be replaced by Anopheles sp. Wolbachia within filaria-infected mosquitoes in our locality gives a hope to use bacteria as a new control trend simultaneously targeting the vector and filarial parasites.

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Faramarz Bozorg-Omid, Mohammad Ali Oshaghi, Mozaffar Vahedi, Fateh Karimian, Seyyed Javad Seyyed-Zadeh, Ali Reza Chavshin
Applied Entomology and Zoology.2020; 55(1): 105. CrossRef
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Brief Communication

Prevalence of Trichomonas vaginalis in Women Visiting 2 Obstetrics and Gynecology Clinics in Daegu, South Korea: Youn-Kyoung Goo, Won-Sik Shin, Hye-Won Yang, So-Young Joo, Su-Min Song, Jae-Sook Ryu, Won-Myung Lee, Hyun-Hee Kong, Won-Ki Lee, Sang-Eun Lee, Won-Ja Lee, Dong-Il Chung, Yeonchul Hong; Korean J Parasitol 2016;54(1):75-80.
Published online February 26, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.1.75

Abstract
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This study explored epidemiological trends in trichomoniasis in Daegu, South Korea. Wet mount microscopy, PCR, and multiplex PCR were used to test for Trichomonas vaginalis in vaginal swab samples obtained from 621 women visiting 2 clinics in Daegu. Of the 621 women tested, microscopy detected T. vaginalis in 4 (0.6%) patients, PCR detected T. vaginalis in 19 (3.0%) patients, and multiplex PCR detected T. vaginalis in 12 (1.9%) patients. Testing via PCR demonstrated high sensitivity and high negative predictive value for T. vaginalis. Among the 19 women who tested positive for T. vaginalis according to PCR, 94.7% (18/19) reported vaginal signs and symptoms. Notably, more than 50% of T. vaginalis infections occurred in females younger than 30 years old, and 58% were unmarried. Multiplex PCR, which simultaneously detects pathogens from various sexually transmitted infections, revealed that 91.7% (11/12) of patients were infected with 2 or more pathogens. Mycoplasma hominis was the most prevalent co-infection pathogen with T. vaginalis, followed by Ureaplasma urealyticum and Chlamydia trachomatis. Our results indicate that PCR and multiplex PCR are the most sensitive tools for T. vaginalis diagnosis, rather than microscopy which has been routinely used to detect T. vaginalis infections in South Korea. Therefore, clinicians should take note of the high prevalence of T. vaginalis infections among adolescent and young women in order to prevent persistent infection and transmission of this disease.

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Original Article

Xenomonitoring of Different Filarial Nematodes Using Single and Multiplex PCR in Mosquitoes from Assiut Governorate, Egypt: Ahmed Kamal Dyab, Lamia Ahmed Galal, Abeer El-Sayed Mahmoud, Yasser Mokhtar; Korean J Parasitol 2015;53(1):77-83.
Published online February 27, 2015; DOI: https://doi.org/10.3347/kjp.2015.53.1.77

Abstract
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Dirofilaria immitis, Dirofilaria immitis, and Dirofilaria repens are filarial nematodes transmitted by mosquitoes belonging to Culex, Aedes, and Anopheles genera. Screening by vector dissection is a tiresome technique. We aimed to screen filarial parasites in their vectors by single and multiplex PCR and evaluate the usefulness of multiplex PCR as a rapid xenomonitoring and simultaneous differentiation tool, in area where 3 filarial parasites are coexisting. Female mosquitoes were collected from 7 localities in Assiut Governorate, were microscopically identified and divided into pools according to their species and collection site. Detection of W. bancrofti, D. immitis, and D. repens using single PCR was reached followed by multiplex PCR. Usefulness of multiplex PCR was evaluated by testing mosquito pools to know which genera and species are used by filarial parasites as a vector. An overall estimated rate of infection (ERI) in mosquitoes was 0.6%; the highest was Culex spp. (0.47%). W. bancrofti, D. immitis, and D. repens could be simultaneously and differentially detected in infected vectors by using multiplex PCR. Out of 100 mosquito pools, 8 were positive for W. bancrofti (ERI of 0.33%) and 3 pools each were positive for D. immitis and D. repens (ERI 0.12%). The technique showed 100% sensitivity and 98% specificity. El-Nikhila, El-Matiaa villages, and Sahel Seleem district in Assiut Governorate, Egypt are still endemic foci for filarial parasites. Multiplex PCR offers a reliable procedure for molecular xenomonitoring of filariasis within their respective vectors in endemic areas. Therefore, it is recommended for evaluation of mosquito infection after lymphatic filariasis eradication programs.

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Brief Communications

Prevalence of Trichomonas vaginalis by PCR in Men Attending a Primary Care Urology Clinic in South Korea: Jun-Hyeok Seo, Hye-Won Yang, So-Young Joo, Su-Min Song, Yu-Ran Lee, Jae-Sook Ryu, Eun Sang Yoo, Won Kee Lee, Hyun-Hee Kong, Sang-Eun Lee, Won-Ja Lee, Youn-Kyoung Goo, Dong-Il Chung, Yeonchul Hong; Korean J Parasitol 2014;52(5):551-555.
Published online October 22, 2014; DOI: https://doi.org/10.3347/kjp.2014.52.5.551

Abstract
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Trichomonas vaginalis, a causative agent of trichomoniasis, may trigger symptomatic or asymptomatic nongonococcal urethritis and chronic prostatitis in men. Despite the availability of highly sensitive diagnostic tests, such as nucleic acid amplification tests, including PCR, few prospective studies present data on male T. vaginalis infection in South Korea. In the present study, the prevalence of T. vaginalis and associated clinical conditions were evaluated in 201 male patients from a primary care urology clinic in South Korea. The prevalence of T. vaginalis infection in our cohort was 4% (8/201) by PCR. T. vaginalis infection was common in men older than 40 years (median age, 52 years). Among the 8 Trichomonas-positive patients, 87.5% (7/8) had prostatic diseases, such as prostatitis and benign prostatic hyperplasia, and 25.0% (2/8) and 12.5% (1/8) were coinfected with Chlamydia trachomatis and Mycoplasma genitalium, respectively. Our results suggest that T. vaginalis infection is not rare in men attending primary care urology clinics in South Korea, especially in those older than 40 years, in whom it may explain the presence of prostatic disease. The possibility of T. vaginalis infection should be routinely considered in older male patients with prostatic diseases in South Korea.

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Systematic review and meta-analysis of the global prevalence and infection risk factors of Trichomonas vaginalis
Wenjie Tian, Yuhua Li, Yani Zhang, Yiming Zhang, Yiran Qin, Yalin Han, Dongxian Li, Shuai Wang, Zhenke Yang, Xiaowei Tian, Xuefang Mei, Zhenchao Zhang
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The Korean Journal of Parasitology.2019; 57(1): 27. CrossRef
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Sun Huh
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Prevalence of Trichomonas vaginalis in Women Visiting 2 Obstetrics and Gynecology Clinics in Daegu, South Korea
Youn-Kyoung Goo, Won-Sik Shin, Hye-Won Yang, So-Young Joo, Su-Min Song, Jae-Sook Ryu, Won-Myung Lee, Hyun-Hee Kong, Won-Ki Lee, Sang-Eun Lee, Won-Ja Lee, Dong-Il Chung, Yeonchul Hong
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Recent Situation of Taeniasis in Mongolia (2002-2012): Anu Davaasuren, Temuulen Dorjsuren, Tetsuya Yanagida, Yasuhito Sako, Kazuhiro Nakaya, Abmed Davaajav, Gurbadam Agvaandaram, Tsatsral Enkhbat, Battsetseg Gonchigoo, Nyamkhuu Dulmaa, Gantigmaa Chuluunbaatar, Akira Ito; Korean J Parasitol 2014;52(2):211-214.
Published online April 18, 2014; DOI: https://doi.org/10.3347/kjp.2014.52.2.211

Abstract
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Epidemiological situation of taeniasis in Mongolia was assessed based on mitochondrial DNA identification of the parasite species. Multiplex PCR was used on a total of 194 proglottid specimens of Taenia species and copro-PCR and loop-mediated isothermal amplification (LAMP) assays were utilized for detection of copro-DNA of 37 fecal samples from taeniasis patients submitted to the Mongolian National Center for Communicable Diseases (NCCD) from 2002 to 2012. In addition, 4 out of 44 calcified cysts in beef kept in formalin since 2003 were evaluated for histopathological confirmation of cattle cysticercosis. All proglottid specimens and stool samples were confirmed to be Taenia saginata by multiplex PCR and by copro-PCR and LAMP, respectively. Cysts collected from cattle were morphologically confirmed to be metacestodes of Taenia species. T. saginata taeniasis was identified from almost all ages from a 2-year-old boy up to a 88-year-old woman and most prominently in 15-29 age group (37%, 74/198) followed by 30-44 age group (34.8%, 69/198 ) from 15 of Mongolia's 21 provinces, while cattle cysticerci were found from 12 provinces. The highest proportion of taeniasis patients was in Ulaanbaatar, the capital of Mongolia.

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Sayed Ajmal Qurishi, Hong-Bin Yan, Li Li, John Aeskhaen Ohiolei, Mughees Aizaz Alvi, Lin-Sheng Zhang, Ha Da, Hong-Mei Qiao, Nigus Abebe Shumuye, Bao Hua, Bing-Xin Bai, Wen-Jun Tian, Ju-Mei Xu, Bao-Quan Fu, Wan-Zhong Jia
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Akira Ito, Tiaoying Li, Toni Wandra, Paron Dekumyoy, Tetsuya Yanagida, Munehiro Okamoto, Christine M Budke
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Neurocysticercosis: A case study of a Mongolian traveler who visited China and India with an updated review in Asia
Anu Davaasuren, Abmed Davaajav, Baigalmaa Ukhnaa, Altantsetseg Purvee, Saraa Unurkhaan, Amartuvshin Luvsan, Jenae E. Logan, Akira Ito
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Articles from Symposium on Asian Taenia (October 2011, Osong, Korea)

Molecular Approaches to Taenia asiatica: Hyeong-Kyu Jeon, Keeseon S. Eom; Korean J Parasitol 2013;51(1):1-8.
Published online February 18, 2013; DOI: https://doi.org/10.3347/kjp.2013.51.1.1

Abstract
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Taenia solium, T. saginata, and T. asiatica are taeniid tapeworms that cause taeniasis in humans and cysticercosis in intermediate host animals. Taeniases remain an important public health concerns in the world. Molecular diagnostic methods using PCR assays have been developed for rapid and accurate detection of human infecting taeniid tapeworms, including the use of sequence-specific DNA probes, PCR-RFLP, and multiplex PCR. More recently, DNA diagnosis using PCR based on histopathological specimens such as 10% formalin-fixed paraffin-embedded and stained sections mounted on slides has been applied to cestode infections. The mitochondrial gene sequence is believed to be a very useful molecular marker for not only studying evolutionary relationships among distantly related taxa, but also for investigating the phylo-biogeography of closely related species. The complete sequence of the human Taenia tapeworms mitochondrial genomes were determined, and its organization and structure were compared to other human-tropic Taenia tapeworms for which complete mitochondrial sequence data were available. The multiplex PCR assay with the Ta4978F, Ts5058F, Tso7421F, and Rev7915 primers will be useful for differential diagnosis, molecular characterization, and epidemiological surveys of human Taenia tapeworms.

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Original Article

Morphologic and Genetic Identification of Taenia Tapeworms in Tanzania and DNA Genotyping of Taenia solium: Keeseon S. Eom, Jong-Yil Chai, Tai-Soon Yong, Duk-Young Min, Han-Jong Rim, Charles Kihamia, Hyeong-Kyu Jeon; Korean J Parasitol 2011;49(4):399-403.
Published online December 16, 2011; DOI: https://doi.org/10.3347/kjp.2011.49.4.399

Abstract
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Species identification of Taenia tapeworms was performed using morphologic observations and multiplex PCR and DNA sequencing of the mitochondrial cox1 gene. In 2008 and 2009, a total of 1,057 fecal samples were collected from residents of Kongwa district of Dodoma region, Tanzania, and examined microscopically for helminth eggs and proglottids. Of these, 4 Taenia egg positive cases were identified, and the eggs were subjected to DNA analysis. Several proglottids of Taenia solium were recovered from 1 of the 4 cases. This established that the species were T. solium (n=1) and T. saginata (n=3). One further T. solium specimen was found among 128 fecal samples collected from Mbulu district in Arusha, and this had an intact strobila with the scolex. Phylegenetic analysis of the mtDNA cox1 gene sequences of these 5 isolates showed that T. saginata was basal to the T. solium clade. The mitochondrial cox1 gene sequences of 3 of these Tanzanian isolates showed 99% similarity to T. saginata, and the other 2 isolates showed 100% similarity to T. solium. The present study has shown that Taenia tapeworms are endemic in Kongwa district of Tanzania, as well as in a previously identified Mbulu district. Both T. solium isolates were found to have an "African/Latin American" genotype (cox1).

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Brief Communication

Molecular Identification of Taenia Tapeworms by Cox1 Gene in Koh Kong, Cambodia: Hyeong-Kyu Jeon, Tai-Soon Yong, Woon-Mok Sohn, Jong-Yil Chai, Sung-Jong Hong, Eun-Taek Han, Hoo-Gn Jeong, Tep Chhakda, Muth Sinuon, Duong Socheat, Keeseon S. Eom; Korean J Parasitol 2011;49(2):195-197.
Published online June 14, 2011; DOI: https://doi.org/10.3347/kjp.2011.49.2.195

Abstract
PDF

We collected fecal samples from 21 individuals infected with Taenia tapeworms in Koh Kong Province, Cambodia, and performed nucleotide sequencing of the cox1 gene and multiplex PCR on the eggs for DNA differential diagnosis of human Taenia tapeworms. Genomic DNA was extracted from the eggs of a minimum number of 10 isolated from fecal samples. Using oligonucleotide primers Ta7126F, Ts7313F, Tso7466F, and Rev7915, the multiplex PCR assay proved useful for differentially diagnosing Taenia solium, Taenia saginata, and Taenia asiatica based on 706, 629, and 474 bp bands, respectively. All of the Taenia specimens from Kho Kong, Cambodia, were identified as either T. saginata (n=19) or T. solium (n=2) by cox1 sequencing and multiplex PCR.

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Original Article

Multiplex PCR Detection of Waterborne Intestinal Protozoa: Microsporidia, Cyclospora, and Cryptosporidium: Seung-Hyun Lee, Migyo Joung, Sejoung Yoon, Kyoungjin Choi, Woo-Yoon Park, Jae-Ran Yu; Korean J Parasitol 2010;48(4):297-301.
Published online December 16, 2010; DOI: https://doi.org/10.3347/kjp.2010.48.4.297

Abstract
PDF

Recently, emerging waterborne protozoa, such as microsporidia, Cyclospora, and Cryptosporidium, have become a challenge to human health worldwide. Rapid, simple, and economical detection methods for these major waterborne protozoa in environmental and clinical samples are necessary to control infection and improve public health. In the present study, we developed a multiplex PCR test that is able to detect all these 3 major waterborne protozoa at the same time. Detection limits of the multiplex PCR method ranged from 10¹ to 10² oocysts or spores. The primers for microsporidia or Cryptosporidium used in this study can detect both Enterocytozoon bieneusi and Encephalitozoon intestinalis, or both Cryptosporidium hominis and Cryptosporidium parvum, respectively. Restriction enzyme digestion of PCR products with BsaBI or BsiEI makes it possible to distinguish the 2 species of microsporidia or Cryptosporidium, respectively. This simple, rapid, and cost-effective multiplex PCR method will be useful for detecting outbreaks or sporadic cases of waterborne protozoa infections.

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Mini Review

Geographical Distribution of Taenia asiatica and Related Species: Keeseon S. Eom, Hyeong-Kyu Jeon, Han-Jong Rim; Korean J Parasitol 2009;47(Suppl):S115.
Published online October 27, 2009; DOI: https://doi.org/10.3347/kjp.2009.47.S.S115

Abstract
PDF

Geographical information of Taenia asiatica is reviewed together with that of T. solium and T. saginata. Current distribution of T. asiatica was found to be mostly from Asian countries: the Republic of Korea, China, Taiwan, Indonesia, and Thailand. Molecular genotypic techniques have found out more countries with T. asiatica from Japan, the Philippines, and Vietnam. Specimens used in this paper were collected from around the world and mostly during international collaboration projects of Korean foundations for parasite control activities (1995-2009) in developing countries.

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