Parasites, Hosts and Diseases

"Tong Soo Kim"

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
PDF

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.

Citations

Citations to this article as recorded by

A rapid and ultrasensitive CRISPR/Cas12a-based assay for the accurate identification of T-even type phages
Chenhang Jiang, Yang Li, Ping Yu, Mengjun Fang, Di Huang, Xiangming Fang, Zhinan Xu
Biotechnology Letters.2025;[Epub] CrossRef
Assessment of Cryptosporidium spp. Sub-Families and Giardia duodenalis Assemblages A and B in Ghanaian HIV Patients, Including Socio-Economic, Clinical, and Immunological Associations
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
Infectious Disease Reports.2025; 17(5): 129. CrossRef
Evaluation of molecular-based methods for the detection and quantification of Cryptosporidium spp. in wastewater
Oumaima Hachimi, Rebecca Falender, Gabriel Davis, Rispa Vranka Wafula, Melissa Sutton, June Bancroft, Paul Cieslak, Christine Kelly, Devrim Kaya, Tyler Radniecki
Science of The Total Environment.2024; 947: 174219. CrossRef
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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
The Importance of Endoscopy with Biopsy: Real-World Evidence of Gastrointestinal Involvement in Primary Immunodeficiency in Two Main Northern Italian Centres
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
Ukraïnsʹkij žurnal medicini, bìologìï ta sportu.2019; 4(2): 149. CrossRef
Molecular epidemiology of Giardia and Cryptosporidium infections – What's new?
R.C.A. Thompson, A. Ash
Infection, Genetics and Evolution.2019; 75: 103951. CrossRef
Cyclospora cayetanensis and Cyclosporiasis: An Update
Sonia Almeria, Hediye N. Cinar, Jitender P. Dubey
Microorganisms.2019; 7(9): 317. CrossRef

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294 Download
19 Web of Science
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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.

Citations

Citations to this article as recorded by

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
Journal of Dairy Science.2021; 104(5): 5152. CrossRef
Sporadic cyclosporiasis in symptomatic Cuban patients: Confirmation of positive results from conventional diagnostic methods by molecular assay
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
Cyclospora cayetanensis and Cyclosporiasis: An Update
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
Entamoeba histolytica: an Overview
Nadia A. El-Dib
Current Tropical Medicine Reports.2017; 4(1): 11. CrossRef
Cryptosporidium and Cyclospora Diarrheal Infection in Malnourished Children: a Nutritional Approach
María A. Echevarría, María L. Eva
Current Tropical Medicine Reports.2017; 4(3): 178. CrossRef

12,396 View
260 Download
9 Web of Science
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Experimental infection of Anopheles sinensis with Korean isolates of Plasmodium vivax: Hyeong Woo Lee, Shin Hyeong Cho, E-Hyun Shin, Jong Soo Lee, Joon-Sang Lee, Jong-Yil Chai, Soon-Hyung Lee, Tong Soo Kim; Korean J Parasitol 2001;39(2):177-183.
Published online June 30, 2001; DOI: https://doi.org/10.3347/kjp.2001.39.2.177

Abstract
PDF

The
objective
s of the present study were to (1) determine the susceptibility of Anopheles sinensis to Korean isolates of Plasmodium vivax, (2) establish a method to collect large quantities of P. vivax sporozoites for use as antigen in seroepidemiological studies, and (3) investigate the characteristics of Korean isolates of P. vivax sporozoites. Females of Anopheles sinensis were collected at non-epidemic area, Seokwha-ri, Cheongwon-gun and Chungcheongbuk-do using tent-trap methods coupled with dry ice. The females were artificially infected with gametocytes of P. vivax using blood obtained from P. vivax malaria patients. Individual mosquitoes were infected using either a parafilm-covered glass feeding apparatus or were allowed to feed on naturally infected volunteers. Mosquitoes were sacrificed between 16 and 18 days post-feeding and an enzyme-linked immunosorbent assay (ELISA) was used to detect sporozoites. Four (33.4%) of 12 mosquitoes, which were fed on naturally infected volunteers directly, were positive for sporozoites. In cases, the mosquitoes allowed to feed on whole blood which were extract from three different patients with heparin treated vacuutainers using a parafilm-covered glass apparatus. Two of 55 (3.6%) were positive which blood sample was maintained at room temperature for 8 hours, 1 of 68 (1.5%) was positive which blood was maintained at 4℃ for 24 hours and 1 of 47 (2.3%) was positive at 4℃ for 48 hours. The mean number of sporozoites was estimated about 818 (n=8; range of 648-1,056) based on optical density values of ELISA.

Citations

Citations to this article as recorded by

국내 말라리아 매개모기 집단유전학 분석
하늘 정, 보경 한, 현일 신, 명노 이, 정원 주, 희일 이
Public Health Weekly Report.2025; 18(16): 623. CrossRef
Genetic diversity and spatiotemporal population structure of Anopheles sinensis in the Republic of Korea based on the mitochondrial cytochrome c oxidase subunit I (COI) marker
Haneul Jung, Bo Gyeong Han, Hyun-Il Shin, Myoung-Ro Lee, Jung-Won Ju, Hee-Il Lee
Osong Public Health and Research Perspectives.2025; 16(4): 348. CrossRef
Plasmodium vivax gametocytes and transmission
Sirasate Bantuchai, Hisham Imad, Wang Nguitragool
Parasitology International.2022; 87: 102497. CrossRef
Quality Change of Fermented Soybean Products by Aspergillus spp. from Soybean Cultivar
Dong Sun Shin, In Duck Choi, Seuk Ki Lee, Ji Young Park, Nam Geol Kim, Kwang-Ho Jeong, Chang Hwan Park, Hye Sun Choi
Food Engineering Progress.2019; 23(4): 258. CrossRef
Epidemiological Characteristics of Re-emerging Vivax Malaria in the Republic of Korea (1993-2017)
Young Yil Bahk, Hyeong-Woo Lee, Byoung-Kuk Na, Jeonga Kim, Kyoung Jin, Yeong Seon Hong, Tong-Soo Kim
The Korean Journal of Parasitology.2018; 56(6): 531. CrossRef
Singapore’s Anopheles sinensis Form A is susceptible to Plasmodium vivax isolates from the western Thailand–Myanmar border
Sook-Cheng Pang, Chiara Andolina, Benoit Malleret, Peter R. Christensen, Sai-Gek Lam-Phua, Muhammad Aliff Bin Abdul Razak, Chee-Seng Chong, Daiqin Li, Cindy S. Chu, Bruce Russell, Laurent Rénia, Lee-Ching Ng, Francois Nosten
Malaria Journal.2017;[Epub] CrossRef
The dominant Anopheles vectors of human malaria in the Asia-Pacific region: occurrence data, distribution maps and bionomic précis
Marianne E Sinka, Michael J Bangs, Sylvie Manguin, Theeraphap Chareonviriyaphap, Anand P Patil, William H Temperley, Peter W Gething, Iqbal RF Elyazar, Caroline W Kabaria, Ralph E Harbach, Simon I Hay
Parasites & Vectors.2011;[Epub] CrossRef
The role of Pvs28 in sporozoite development in Anopheles sinensis and its longevity in BALB/c mice
Tong-Soo Kim, Hyung-Hwan Kim, Sung-Ung Moon, Sun-Sim Lee, E-Hyun Shin, Chang-Mi Oh, Yoon-Joong Kang, Do-Kyung Kim, Youngjoo Sohn, Hyuck Kim, Hyeong-Woo Lee
Experimental Parasitology.2011; 127(2): 346. CrossRef
The susceptibility of Anopheles lesteri to infection with Korean strain of Plasmodium vivax
Deepak Joshi, Wej Choochote, Mi-Hyun Park, Jung-Yeon Kim, Tong-Soo Kim, Wannapa Suwonkerd, Gi-Sik Min
Malaria Journal.2009;[Epub] CrossRef
Anopheles kleini,Anopheles pullus, andAnopheles sinensis: Potential Vectors ofPlasmodium vivaxin the Republic of Korea
Won-Ja Lee, Terry A. Klein, Heung-Chul Kim, Young-Mi Choi, Seok-Hyun Yoon, Kyu-Sik Chang, Sung-Tae Chong, In-Yong Lee, James W. Jones, Jolanta S. Jacobs, Jetsumon Sattabongkot, Jae-Sun Park
Journal of Medical Entomology.2007; 44(6): 1086. CrossRef
Anopheles kleini, Anopheles pullus, and Anopheles sinensis: Potential Vectors of Plasmodium vivax in the Republic of Korea
Won-Ja Lee, Terry A. Klein, Heung-Chul Kim, Young-Mi Choi, Seok-Hyun Yoon, Kyu-Sik Chang, Sung-Tae Chong, In-Yong Lee, James W. Jones, Jolanta S. Jacobs, Jetsumon Sattabongkot, Jae-Sun Park
Journal of Medical Entomology.2007; 44(6): 1086. CrossRef
Detection of vivax sporozoites naturally infected in Anopheline mosquitoes from endemic areas of northern parts of Gyeonggi-do (province) in Korea
Hyeong Woo Lee, E Hyun Shin, Shin Hyeong Cho, Hee Il Lee, Chung Lim Kim, Wook Gyo Lee, Sung Ung Moon, Jong Soo Lee, Wan Ja Lee, Tong Soo Kim
The Korean Journal of Parasitology.2002; 40(2): 75. CrossRef
Vector competence of Anopheles lesteri Baisas and Hu (Diptera: Culicidae) to Plasmodium vivax in Korea
E-Hyun Shin, Tong-Soo Kim, Hyeong-Woo Lee, Jong-Soo Lee, Won-Ja Lee
The Korean Journal of Parasitology.2002; 40(1): 41. CrossRef
Intraspecific Hybridization of Anopheles sinensis (Diptera: Culicidae) Strains from Thailand and Korea
Gi-Sik Min, Wej Choochote, Atchariya Jitpakdi, Se-Joo Kim, Won Kim, Jongwoo Jung, Anuluck Junkum
Molecules and Cells.2002; 14(2): 198. CrossRef

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Western blot diagnosis of vivax malaria with multiple stage-specific antigens of the parasite: Eui-Sun Son, Tong Soo Kim, Ho-Woo Nam; Korean J Parasitol 2001;39(2):171-176.
Published online June 30, 2001; DOI: https://doi.org/10.3347/kjp.2001.39.2.171

Abstract
PDF

Western blot analysis was performed to diagnose vivax malaria using stage-specific recombinant antigens. Genomic DNA from the whole blood of a malaria patient was used as templates to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), merozoite surface protein (MSP-1), apical merozoite antigen (AMA-1), serine repeat antigen (SERA), and exported antigen (EXP-1) of Plasmodium vivax. Each amplified DNA fragment was inserted into a pGEX-4T plasmid to induce the expression of GST fusion protein in Escherichia coli by IPTG. The bacterial cell extracts were separated on 10% SDS-PAGE followed by western blot analysis with patient sera which was confirmed by blood smear examination. When applied with patient sera, 147 (91.9%) out of 160 vivax malaria, 12 (92.3%) out of 13 falciparum malaria, and all 9 vivax/falciparum mixed malaria reacted with at least one antigen, while no reactions occurred with 20 normal uninfected sera. In the case of vivax malaria, CSP-1 reacted with 128 (80.0%) sera, MSP-1 with 102 (63.8%), AMA-1 with 128 (80.0%), SERA with 115 (71.9%), and EXP-1 with 89 (55.6%), respectively. We obtained higher detection rates when using 5 antigens (91.9%) rather than using each antigen solely (55.6-80%), a combination of 2 (76.3-87.5%), 3 (85.6-90.6%), or 4 antigens (89.4-91.3%). This method can be applied to serological diagnosis, mass screening in endemic regions, or safety test in transfusion of prevalent vivax malaria.

Citations

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A Dual, Systematic Approach to Malaria Diagnostic Biomarker Discovery
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Frontiers in Cellular and Infection Microbiology.2022;[Epub] CrossRef
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Biosensors.2017; 7(3): 28. CrossRef
Expression and Evaluation of Recombinant Plasmodium knowlesi Merozoite Surface Protein-3 (MSP-3) for Detection of Human Malaria
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ELISA detection of vivax malaria with recombinant multiple stage-specific antigens and its application to survey of residents in endemic areas
Sera Kim, Hye-Jin Ahn, Tong-Soo Kim, Ho-Woo Nam
The Korean Journal of Parasitology.2003; 41(4): 203. CrossRef