Parasites, Hosts and Diseases

"western blot"

Brief Communications

Detection of Human Anti-Trypanosoma cruzi Antibody with Recombinant Fragmented Ribosomal P Protein: Yeong Hoon Kim, Zhaoshou Yang, Jihoo Lee, Hye-Jin Ahn, Chom-Kyu Chong, Wagner Maricondi, Ronaldo F. Dias, Ho-Woo Nam; Korean J Parasitol 2019;57(4):435-437.
Published online August 31, 2019; DOI: https://doi.org/10.3347/kjp.2019.57.4.435

Abstract
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Chagas disease is caused by the protozoan parasite Trypanosoma cruzi, and is endemic in many Latin American countries. Diagnosis is based on serologic testing and the WHO recommends two or more serological tests for confirmation. Acidic ribosomal P protein of T. cruzi showed strong reactivity against positive sera of patients, and we cloned the protein after fragmenting it to enhance its antigenicity and solubility. Twelve positive sera of Chagas disease patients were reacted with the fragmented ribosomal P protein using western blot. Detection rate and density for each fragment were determined. Fragments F1R1, F1R2, and F2R1 showed 100% rate of detection, and average density scoring of 2.00, 1.67, and 2.42 from a maximum of 3.0, respectively. Therefore, the F2R1 fragment of the ribosomal P protein of T. cruzi could be a promising antigen to use in the diagnosis of Chagas disease in endemic regions with high specificity and sensitivity.

Citations

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In vitro diagnostic methods of Chagas disease in the clinical laboratory: a scoping review
Luis C. Ascanio, Savannah Carroll, Alberto Paniz-Mondolfi, Juan David Ramírez
Frontiers in Microbiology.2024;[Epub] CrossRef
A refined genome phage display methodology delineates the human antibody response in patients with Chagas disease
André Azevedo Reis Teixeira, Luis Rodriguez Carnero, Andréia Kuramoto, Fenny Hui Fen Tang, Carlos Hernique Gomes, Natalia Bueno Pereira, Léa Campos de Oliveira, Regina Garrini, Jhonatas Sirino Monteiro, João Carlos Setubal, Ester Cerdeira Sabino, Renata P
iScience.2021; 24(6): 102540. CrossRef

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Western Blot Detection of Human Anti-Chikungunya Virus Antibody with Recombinant Envelope 2 Protein: Zhaoshou Yang, Jihoo Lee, Hye-Jin Ahn, Chom-Kyu Chong, Ronaldo F. Dias, Ho-Woo Nam; Korean J Parasitol 2016;54(2):239-241.
Published online April 30, 2016; DOI: https://doi.org/10.3347/kjp.2016.54.2.239

Abstract
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Chikungunya virus (CHIKV), a tropical pathogen, has re-emerged and has massive outbreaks abruptly all over the world. Containing many dominant epitopes, the envelope E2 protein of CHIKV has been explored for the vaccination or diagnosis. In the present study, the antigenicity of a recombinant expressed intrinsically disorder domain (IUD) of E2 was tested for the detection of the antibody against CHIKV through western blot method. The gene of the IUD of E2 was inserted into 2 different vectors and expressed as recombinant GST-E2 and recombinant MBP-E2 fusion protein, respectively. Two kinds of fusion proteins were tested with 30 CHIKV patient sera and 30 normal sera, respectively. Both proteins were detected by 25 patients sera (83.3%) and 1 normal serum (3.3%). This test showed a relatively high sensitivity and very high specificity of the recombinant E2 proteins to be used as diagnostic antigens against CHIKV infection.

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Anna Andrew, Tholasi Nadhan Navien, Tzi Shien Yeoh, Marimuthu Citartan, Ernest Mangantig, Magdline S. H. Sum, Ewe Seng Ch’ng, Thean-Hock Tang, Guilherme S. Ribeiro
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An Overview of Laboratory Diagnosis of Central Nervous System Viral Infections
Sharifah Aliah Diyanah Syed Hussin, Ang-Lim Chua, Hassanain Al-Talib, Shamala Devi Sekaran, Seok Mui Wang
Journal of Pure and Applied Microbiology.2022; 16(4): 2225. CrossRef
Molecular and Immunological Diagnostic Techniques of Medical Viruses
Daniel Hussien Reta, Tesfaye Sisay Tessema, Addis Simachew Ashenef, Adey Feleke Desta, Wajana Lako Labisso, Solomon Tebeje Gizaw, Solomon Mequanente Abay, Daniel Seifu Melka, Fisseha Alemu Reta
International Journal of Microbiology.2020; 2020: 1. CrossRef
Apigenin enhances the cisplatin cytotoxic effect through p53-modulated apoptosis
Rui Liu, Ping Ji, Bin Liu, Haishi Qiao, Xia Wang, Likun Zhou, Ting Deng, Yi Ba
Oncology Letters.2017; 13(2): 1024. CrossRef

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Case Report

Imported Intraocular Gnathostomiasis with Subretinal Tracks Confirmed by Western Blot Assay: Ji Ho Yang, Moosang Kim, Eung Suk Kim, Byoung-Kuk Na, Seung-Young Yu, Hyung-Woo Kwak; Korean J Parasitol 2012;50(1):73-78.
Published online March 6, 2012; DOI: https://doi.org/10.3347/kjp.2012.50.1.73

Abstract
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We report a case of intraocular gnathostomiasis diagnosed by western blot assay in a patient with subretinal tracks. A 15-year-old male patient complained of blurred vision in the right eye, lasting for 2 weeks. Eight months earlier, he had traveled to Vietnam for 1 week and ate raw wild boar meat and lobster. His best-corrected visual acuity was 20/20 in both eyes and anterior chamber examination revealed no abnormalities. Fundus examination showed subretinal tracks in the right eye. Fluorescein angiography and indocyanine green angiography showed linear hyperfluorescence of the subretinal lesion observed on fundus in the right eye. Ultrasound examination revealed no abnormalities. Blood tests indicated mild eosinophilia (7.5%), and there was no abnormality found by systemic examinations. Two years later, the patient visited our department again for ophthalmologic evaluation. Visual acuity remained 20/20 in both eyes and the subretinal tracks in the right eye had not changed since the previous examination. Serologic examination was performed to provide a more accurate diagnosis, and the patient's serum reacted strongly to the Gnathostoma nipponicum antigen by western blot assay, which led to a diagnosis of intraocular gnathostomiasis. This is the first reported case of intraocular gnathostomiasis with subretinal tracks confirmed serologically using western blot in Korea.

Citations

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Ocular infections in international travelers
Francesca F. Norman, Julio J. González-López, Diego Gayoso-Cantero, Marta Vicente-Antolin, Maria-Dolores Corbacho-Loarte, Rogelio López-Vélez, Marta González-Sanz
Travel Medicine and Infectious Disease.2025; 63: 102789. CrossRef
Imported parasitic diseases in the Republic of Korea: status and issues
Jong-Yil Chai
Journal of the Korean Medical Association.2025; 68(1): 52. CrossRef
Intracameral gnathostomiasis presenting as neuroretinitis
Kimhour Lay, Leng Un, Kossama Chukmol, Piseth Kong
BMJ Case Reports.2025; 18(3): e260393. CrossRef
Gastrodin reduces Aβ brain levels in an Alzheimer's disease mouse model by inhibiting P-glycoprotein ubiquitination
Chenghao Zhu, Shangtao Wang, Siyu Ma, Shurui Zhang, Yanjun Wang, Baoshan Li, Wei Zhang, Zhirong Sun
Phytomedicine.2024; 135: 156229. CrossRef
Intracameral Gnathostomiasis: A Case Report and Literature Review
Wijak Kongwattananon, Thanaporn Wiriyabanditkul, Waraluck Supwatjariyakul, Thanapong Somkijrungroj
Ocular Immunology and Inflammation.2023; 31(5): 1092. CrossRef
A case of vocal cord gnathostomiasis diagnosed with sectional morphologies in a histopathological specimen from a Chinese woman living in Korea
Doo Sik Park, Eun Hyun Cho, Kyung Hoon Park, Soo Min Jo, Bumjung Park, Sun Huh
Parasites, Hosts and Diseases.2023; 61(3): 298. CrossRef
Subretinal gnathostomiasis: A successful focal laser photocoagulation for a living parasite
Supalert Prakhunhungsit, Somanus Thoongsuwan, Sutasinee Boonsopon, Thitiyaporn Panawattanawong, Pitchaya Amornvararak, Nuttawut Rodanant, Nopasak Phasukkijwatana
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Jorge Costa Eiras, Gilberto Cezar Pavanelli, Ricardo Massato Takemoto, Yukifumi Nawa
The Korean Journal of Parasitology.2018; 56(3): 215. CrossRef
Imported gnathostomiasis manifesting as cutaneous larva migrans and Löffler’s syndrome
William L. Hamilton, Daniel Agranoff
BMJ Case Reports.2018; 2018: bcr-2017-223132. CrossRef
Ocular Gnathostomiasis—Update of Earlier Survey
Kittisak Sawanyawisuth, Yukifumi Nawa, Sandra Fernández Figueiras, Sylvia Paz Diaz Camacho, Verajit Chotmongkol, Masahide Yoshikawa, Maria Benavides
The American Journal of Tropical Medicine and Hygiene.2017; 97(4): 1232. CrossRef
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S. Hem, A. Tarantola, R. Chheang, P. Nop, A. Kerléguer
Bulletin de la Société de pathologie exotique.2015; 108(5): 312. CrossRef
Three-dimensional spectral domain optical coherence tomography and light microscopy of an intravitreal parasite
Aziz A. Khanifar, Michael J. Espiritu, Jane S. Myung, Grant D. Aaker, Audrey N. Schuetz, Donald J. D’Amico, R. V. Paul Chan
Journal of Ophthalmic Inflammation and Infection.2015;[Epub] CrossRef
Toxocara Optic Neuropathy after Ingestion of Raw Meat Products
Hee Kyung Yang, Se Joon Woo, Jeong-Min Hwang
Optometry and Vision Science.2014; 91(11): e267. CrossRef
Gnathostoma spinigerum Infection in the Upper Lip of a Korean Woman: An Autochthonous Case in Korea
Jae Hee Kim, Hyemi Lim, Young-Sang Hwang, Tae Yeon Kim, Eun Mee Han, Eun-Hee Shin, Jong-Yil Chai
The Korean Journal of Parasitology.2013; 51(3): 343. CrossRef
Cutaneous Gnathostomiasis with Recurrent Migratory Nodule and Persistent Eosinophilia: a Case Report from China
Jing Cui, Ye Wang, Zhong Quan Wang
The Korean Journal of Parasitology.2013; 51(4): 467. CrossRef

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Original Articles

Seroprevalence of Antibodies against Anisakis simplex Larvae among Health-Examined Residents in Three Hospitals of Southern Parts of Korea: Jung Kim, Jin Ok Jo, Seon Hee Choi, Min Kyoung Cho, Hak Sun Yu, Hee Jae Cha, Meesun Ock; Korean J Parasitol 2011;49(2):139-144.
Published online June 14, 2011; DOI: https://doi.org/10.3347/kjp.2011.49.2.139

Abstract
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The present study was performed to estimate the seroprevalence of larval Anisakis simplex infection among the residents health-examined in 3 hospitals in southern parts of Korea. A total of 498 serum samples (1 serum per person) were collected in 3 hospitals in Busan Metropolitan city, Masan city, and Geoje city in Gyeongsangnam-do (Province) and were examined by IgE-ELISA and IgE-western blotting with larval A. simplex crude extract and excretory-secretory products (ESP). The prevalence of antibody positivity was 5.0% and 6.6% with ELISA against crude extracts and ESP, respectively. It was also revealed that infection occurred throughout all age groups and higher in females than in males. A specific protein band of 130 kDa was detected from 10 patients with western blot analysis against crude extract and ESP among those who showed positive results by ELISA. Our study showed for the first time the seroprevalence of anisakiasis in Korea. The allergen of 130 kDa can be a candidate for serologic diagnosis of anisakiasis.

Citations

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Seroprevalence of IgG and IgE Antibodies Against Anisakis in the Presumably Healthy Population of the Canary Islands
Eligia González-Rodríguez, Marta Rodero, J. Alberto Montoya-Alonso, Kevin M. Santana-Hernández, Myriam R. Ventura, Carmen Cuéllar, Eligia Rodríguez-Ponce
Antibodies.2025; 14(3): 60. CrossRef
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Shin Ae Kang, Hak Sun Yu
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Progress in Anisakis Allergy Research: Milestones and Reversals
Alvaro Daschner, Carmen Cuéllar
Current Treatment Options in Allergy.2020; 7(4): 457. CrossRef
IgE sensitization to Anisakis pegreffii in Italy: Comparison of two methods for the diagnosis of allergic anisakiasis
S. Mattiucci, A. Colantoni, B. Crisafi, F. Mori‐Ubaldini, L. Caponi, P. Fazii, G. Nascetti, F. Bruschi
Parasite Immunology.2017;[Epub] CrossRef
Anthelmintic Activities of Aporphine from Nelumbo nucifera Gaertn. cv. Rosa-plena against Hymenolepis nana
Rong-Jyh Lin, Mei-Hsuan Wu, Yi-Hsuan Ma, Li-Yu Chung, Chung-Yi Chen, Chuan-Min Yen
International Journal of Molecular Sciences.2014; 15(3): 3624. CrossRef
Endoscopic imaging of parasites in the human digestive tract
Naoki Hosoe, Haruhiko Ogata, Toshifumi Hibi
Parasitology International.2014; 63(1): 216. CrossRef
An Extended Study of Seroprevalence of Anti‐Anisakis simplex IgE Antibodies in Norwegian Blood Donors
A. H. Lin, I. Nepstad, E. Florvaag, E. Egaas, T. Van Do
Scandinavian Journal of Immunology.2014; 79(1): 61. CrossRef
Occurrence of anisakid nematode larvae in chub mackerel (Scomber japonicus) caught off Korea
Tae-Jong Bak, Chan-Hyeok Jeon, Jeong-Ho Kim
International Journal of Food Microbiology.2014; 191: 149. CrossRef
Clinical Characteristics of Gastroallergic Anisakiasis and Diagnostic Implications of Immunologic Tests
Young-Bae Chung, Jaechun Lee
Allergy, Asthma & Immunology Research.2014; 6(3): 228. CrossRef
Preventive and Therapeutic Effects of <italic>Anisakis simplex</italic> Larval Protein in a Mouse Model of Crohn’S Disease
Hee-Jae Cha, Mee Sun Ock
Kosin Medical Journal.2013; 28(2): 107. CrossRef
Cross-sectional study of serum reactivity to Anisakis simplex in healthy adults in Niter?i, Brazil
Israel Junior, Mauricio Vericimo, Luciana Cardoso, Sergio Clemente, Elmiro Nascimento, Gerlinde Teixeira
Acta Parasitologica.2013;[Epub] CrossRef
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Jeannette Guarner
Scientifica.2012; 2012: 1. CrossRef
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Hee Jae Cha, Mee Sun Ock
Kosin Medical Journal.2012; 27(2): 73. CrossRef
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Erica L. Pufall, Andria Jones-Bitton, Scott A. McEwen, Tanya M. Brown, Victoria L. Edge, Jerzy Rokicki, Katarzyna Karpiej, Andrew S. Peregrine, Manon Simard
Foodborne Pathogens and Disease.2012; 9(11): 1002. CrossRef
A technique for the intra-gastric administration of live larvae of Anisakis simplex in mice
Israel Figueiredo, Luciana Cardoso, Gerlinde Teixeira, Leila Lopes, Sergio Carmona São Clemente, Mauricio Afonso Vericimo
Experimental Parasitology.2012; 130(3): 285. CrossRef

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A Hospital-Based Serological Survey of Cryptosporidiosis in the Republic of Korea: Jong-Kyu Lee, Eun-Taek Han, Sun Huh, Woo-Yoon Park, Jae-Ran Yu; Korean J Parasitol 2009;47(3):219-225.
Published online August 28, 2009; DOI: https://doi.org/10.3347/kjp.2009.47.3.219

Abstract
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The seroprevalence of cryptosporidiosis was examined using patients' sera collected from hospitals located in 4 different areas of the Republic of Korea. ELISA was used to measure antibody titers against Cryptosporidium parvum antigens from a total of 2,394 serum samples, which were collected randomly from patients in local hospitals; 1) Chungbuk National University Hospital, 2) Konkuk University Hospital, 3) local hospitals in Chuncheon, Gangwon-do (province), 4) Jeonnam National University Hospital, from 2002 through 2003. Of the 2,394 samples assayed, 34%, 26%, and 56% were positive for C. parvum-specific IgG, IgM, and IgA antibodies, respectively. Positive IgG titers were most common in sera from Jeonnam National University Hospital, Gwangju, Jeollanam-do, and positive IgM titers were most common in sera from Chungbuk National University Hospital, Cheongju, Chuncheongbuk-do. The seropositivity was positively correlated with age for both the IgG and IgA antibodies but was negatively correlated with age for the IgM antibodies. Western blotting revealed that 92%, 83%, and 77% of sera positive for IgG, IgM, and IgA ELISA reacted with 27-kDa antigens, respectively. These results suggested that infection with Cryptosporidium in hospital patients occurs more commonly than previously reported in the Republic of Korea.

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Eun-Joo Cho, Jin-Young Yang, Eun-Sook Lee, Se-Chul Kim, So-Yang Cha, Sung-Tek Kim, Man-Ho Lee, Sun-Hee Han, Young-Sang Park
The Korean Journal of Parasitology.2013; 51(4): 461. CrossRef
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Clinical and Vaccine Immunology.2012; 19(6): 849. CrossRef
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Migyo Joung, Sejoung Yoon, Kyungmi Choi, Joung-Yeon Kim, Woo-Yoon Park, Jae-Ran Yu
Experimental Parasitology.2011; 129(4): 331. CrossRef
Cryptosporidiosis
Jae-Ran Yu
Hanyang Medical Reviews.2010; 30(3): 187. CrossRef

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Brief Communication

Seroprevalence of Toxocara antibodies among patients suspected of ocular toxocariasis in Slovenia: Jernej Logar, Barbara ?oba, Aleksandra Kraut, Branka Stirn-Kranjc; Korean J Parasitol 2004;42(3):137-140.
Published online September 20, 2004; DOI: https://doi.org/10.3347/kjp.2004.42.3.137

Abstract
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Ocular toxocariasis named also ocular larva migrans is caused by larvae of the roundworm Toxocara spp. The purpose of this study was to find out the seroprevalence of Toxocara antibodies in patients suspected of ocular toxocariasis. Between January 2001 and December 2003, sera from 239 ocular patients, aged 3 to 80 years, were examined by ELISA and confirmed by Western blot test. Out of the 239 patients, 172 (72%) were seronegative and 67 (28%) were Toxocara seropositive; 95% CI (22-34%). The median age of Toxocara seropositive patients was 37.6 years. There was no significant difference in the number of Toxocara positive sera between the younger age group (≤14 years) and the older age group (>14 years), p>0.05. A high rate of Toxocara seropositivity in ocular patients should alert the ophthalmologists in Slovenia to include toxocariasis in the differential diagnosis of eye diseases more frequently.

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A new ELISA and western blot technique based on recombinant TES antigen and/or larval antigen for the detection of toxocariasis in humans
Marie-Kristin Raulf, Daniela Jordan, Herbert Auer, Jens M. Warnecke, Bernd Lepenies, Christina Strube
Parasitology.2021; 148(3): 333. CrossRef
Seroprevalence of Larval Toxocarosis in the Czech Republic
Katerina Skulinova, Jan Novak, Martin Kasny, Libuse Kolarova
Acta Parasitologica.2020; 65(1): 68. CrossRef
Toxocariasis: critical analysis of serology in patients attending a public referral center for ophthalmology in Brazil
Guita Rubinsky-Elefant, Joyce H. Yamamoto, Carlos E. Hirata, Luiz E. Prestes-Carneiro
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Marta Serrano-Moliner, María Morales-Suarez-Varela, M. Adela Valero
Pathogens and Global Health.2018; 112(5): 249. CrossRef
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M. Rudzińska, B. Kowalewska, K. Sikorska
Parasite Immunology.2017;[Epub] CrossRef
Evaluation of the prevalence and clinical impact of toxocariasis in patients with eosinophilia of unknown origin
Hong-Beum Kim, Jun-Won Seo, Jun-Hyung Lee, Byung-Seok Choi, Sang-Gon Park
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Devika Iddawela, Kiruthiha Ehambaram, Pemindra Bandara
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Vojtech Boldiš, František Ondriska, Eva Špitalská, Katarína Reiterová
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Domenico Otranto, Mark L Eberhard
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A-P. Bellanger, P. Humbert, B. Gavignet, A.D. Deschaseaux, C. Barisien, S. Roussel, L. Millon, F. Aubin, R. Piarroux
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L. E. PRESTES-CARNEIRO, D. H. P. SOUZA, G. C. MORENO, C. TROIANI, V. SANTARÉM, S. C. S. ZAGO, N. A. MIGUEL, S. B. Z. FREITAS, R. FARIA, L. MARTINI, G. RUBINSKY-ELEFANT, A. IHA, A. J. VAZ
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Yong-Hun Kim, Sun Huh, Young-Bae Chung
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Lenneke de Visser, Aniki Rothova, Joke H. de Boer, Anton M. van Loon, Frank T. Kerkhoff, Marijke R. Canninga-van Dijk, Annemarie Y.L. Weersink, Jolanda D.F. de Groot-Mijnes
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Original Article

Identification of newly isolated Babesia parasites from cattle in Korea by using the Bo-RBC-SCID mice: Shin-Hyeong Cho, Tong-Soo Kim, Hyeong-Woo Lee, Masayoshi Tsuji, Chiaki Ishihara, Jong-Taek Kim, Sung-Hwan Wee, Chung-Gil Lee; Korean J Parasitol 2002;40(1):33-40.
Published online March 31, 2002; DOI: https://doi.org/10.3347/kjp.2002.40.1.33

Abstract
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Attempts were made to isolate and identify Korean bovine Babesia parasite. Blood samples were collected from Holstein cows in Korea, and Babesia parasites were propagated in SCID mice with circulating bovine red blood cells for isolation. The isolate was then antigenically and genotypically compared with several Japanese isolates. The Korean parasite was found to be nearly identical to the Oshima strain isolated from Japanese cattle, which was recently designated as Babesia ovata oshimensis n. var. Haemaphysalis longicornis was the most probable tick species that transmited the parasite.

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Jung-Yeon Kim, Shin-Hyeong Cho, Hyun-Na Joo, Masayoshi Tsuji, Sung-Ran Cho, Il-Joong Park, Gyung-Tae Chung, Jung-Won Ju, Hyeng-Il Cheun, Hyeong-Woo Lee, Young-Hee Lee, Tong-Soo Kim
Journal of Clinical Microbiology.2007; 45(6): 2084. CrossRef

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Brief Communication

Enzyme-linked immunosorbent assay for detection of Trichinella spiralis antibodies and the surveillance of selected pig breeding farms in the Republic of Korea: Sung-Hwan Wee, Chung-Gil Lee, Hoo-Don Joo, Yung-Bai Kang; Korean J Parasitol 2001;39(3):261-264.
Published online September 30, 2001; DOI: https://doi.org/10.3347/kjp.2001.39.3.261

Abstract
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Trichinellosis is a parasitic zoonosis of public health importance. It is caused by Trichinella spiralis which has a wide host range including humans. In the present communication, the ELISA technique was employed on a total of 803 blood samples from 7 selected pig breeding farms in 1996 for diagnosis and surveillance of trichinellosis. Out of the entire 803 samples, nine were found to be suspected while one was positive by ELISA. But western blot analyses employed for further confirmation have shown that all of 10 samples did not react to larval excretory-secretory product antigens. These results indicate that pig breeding farms included in the present study are free from trichinellosis. However, it does not mean Korea is free from trichinellosis since human trichinellosis has recently been reported. The necessity of continued surveillance for trichinellosis in both pigs and wild animals was discussed.

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Primary characterization and assessment of a T. spiralis antigen for the detection of Trichinella infection in pigs
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Evaluation of ELISA coupled with Western blot as a surveillance tool for Trichinella infection in wild boar (Sus scrofa)
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Seroprevalence of trichinellosis in domestic animals in northwestern Vietnam
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Original Articles

Determination of antigenic domain in GST fused major surface protein (Nc-p43) of Neospora caninum: Eui-Sun Son, Hye-Jin Ahn, Jae-Hoon Kim, Dae-Yong Kim, Ho-Woo Nam; Korean J Parasitol 2001;39(3):241-246.
Published online September 30, 2001; DOI: https://doi.org/10.3347/kjp.2001.39.3.241

Abstract
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The antigenic domain of the major surface protein (Nc-p43) of Neospora caninum was examined by polymerase chain reaction of its gene fragments and recombinant expression as GST fusion proteins. The fragments of Nc-p43 were as follow: a total open reading frame (OFR), T; OFR without signal sequence and C-terminal hydrophobic sequence, S; N-terminal 2/3 parts of S, A; C-terminal 2/3 parts, P; N-terminal 1/3 part, X; middle 1/3 part, Y; and C-terminal 1/3 part, Z, respectively. The DNA fragments were cloned into pGEX-4T vector. Recombinant plasmids transformed into Escherichia coli of BL21 pLysS (DE3) strain were induced to express GST or GST fused fragments of Nc-p43 such as 69 kDa protein for T, 66 kDa for S, 52 kDa for A, 53 kDa for P, and 40 kDa proteins for X, Y, and Z, respectively in SDS-PAGE. The Nc-p43 fragments of T, S, and P reacted with a bovine serum of neosporosis while those of A, X, Y, and Z together with GST did not in the western blot. These findings suggest that the antigenic domain of Nc-p43 of N. caninum may be localized in the C-terminal 2/3 parts. Together with A19 clone in SAG1 of Toxoplasma gondii (Nam et al., 1996), the P fragment of Nc-p43 could be used as efficient antigens to diagnose and differentiate those infections with both species.

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Molecular characterization of Neospora caninum major antigens NcSAG1 and NcSRS2
Soledad Echeverría, Federico Carrión, Martín Soñora, Andrés Cabrera, Carlos Robello
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Expression ofNeospora caninumNcSRS2 surface protein inPichia pastorisand its application for serodiagnosis ofNeosporainfection
Amanda Fernandes Pinheiro, Sibele Borsuk, Maria Elisabeth Aires Berne, Luciano da Silva Pinto, Renato Andreotti, Talita Roos, Barbara Couto Rollof, Fábio Pereira Leivas Leite
Pathogens and Global Health.2013; 107(3): 116. CrossRef
Induction of Interferon-Gamma (IFN-γ) and T Helper 1 (Th1) Immune Response by Bitter Gourd Extract
Kazunori IKE, Yuko UCHIDA, Tomohiko NAKAMURA, Soichi IMAI
Journal of Veterinary Medical Science.2005; 67(5): 521. CrossRef
ELISA detection of IgG antibody against a recombinant major surface antigen (Nc-p43) fragment of Neospora caninum in bovine sera
Hye-Jin Ahn, Sera Kim, Dae-Yong Kim, Ho-Woo Nam
The Korean Journal of Parasitology.2003; 41(3): 175. 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.

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A Dual, Systematic Approach to Malaria Diagnostic Biomarker Discovery
Seda Yerlikaya, Ewurama D A Owusu, Augustina Frimpong, Robert Kirk DeLisle, Xavier C Ding
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Antigen Discovery in Circulating Extracellular Vesicles From Plasmodium vivax Patients
Iris Aparici-Herraiz, Melisa Gualdrón-López, Carlos J. Castro-Cavadía, Jaime Carmona-Fonseca, María Fernanda Yasnot, Carmen Fernandez-Becerra, Hernando A. del Portillo
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Aver Hemben, Jon Ashley, Ibtisam Tothill
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Jeremy Ryan De Silva, Yee-Ling Lau, Mun-Yik Fong, Luzia Helena Carvalho
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Yee Ling Lau, Fei Wen Cheong, Rohela Mahmud, Mun Yik Fong
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Identification of Plasmodium vivax Proteins with Potential Role in Invasion Using Sequence Redundancy Reduction and Profile Hidden Markov Models
Daniel Restrepo-Montoya, David Becerra, Juan G. Carvajal-Patiño, Alvaro Mongui, Luis F. Niño, Manuel E. Patarroyo, Manuel A. Patarroyo, Leonardo Mariño-Ramírez
PLoS ONE.2011; 6(10): e25189. CrossRef
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

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Detection of IgG antibody against Neospora caninum in cattle in Korea: Ji-Seon Bae, Dae-Yong Kim, Woo-Suk Hwang, Jae-Hoon Kim, Nam-Seok Lee, Ho-Woo Nam; Korean J Parasitol 2000;38(4):245-249.
Published online December 31, 2000; DOI: https://doi.org/10.3347/kjp.2000.38.4.245

Abstract
PDF

A total of 492 cattle sera was screened by IgG-ELISA against Neospora caninum (Nc-1 strain and a Korean isolate, KBA-2) and Toxoplasma gondii. Out of 492, 113 sera (23.0%) reacted positively to either Nc-1 or KBA-2 strains of N. caninum. Among the 113 positive sera, 92 sera (81.4%) reacted with antigens of both strains, but 6 sera (5.3%) with Nc-1 and 15 sera (13.3%) with KBA-2 strain only. And with T. gondii antigen, 6 sera (1.2%) were positive but all reacted with N. caninum antigen also. Western blot revealed typical binding pattern according to ELISA values, such that high OD group reacted specifically to the major surface proteins including 43 kDa protein. Seroprevalence of 23.0% indicates that neosporosis seemed to be one of major causes of abortion in cattle. It is suggested here to establish more epidemiological researches nationwide systematically.

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Serological evaluation of Neospora caninum in pregnant women treated at referral center for prenatal screening in Mato Grosso do Sul, Brazil
Pâmella Oliveira Duarte, Bárbara Guimarães Csordas, Leandra Marla Oshiro, Leandro de Oliveira Souza Higa, Namor Pinheiro Zimmermann, Kauê Rodriguez Martins, Jacqueline Cavalcante Barros, Renato Andreotti
Revista Brasileira de Parasitologia Veterinária.2020;[Epub] CrossRef
Meta-analysis of the prevalence and risk factors associated with bovine neosporosis
Claudia Mello Ribeiro, Isabela Ribeiro Soares, Rodrigo Guerrero Mendes, Paula Andrea de Santis Bastos, Satie Katagiri, Renato Bacarin Zavilenski, Hudson Felipe Porto de Abreu, Vera Afreixo
Tropical Animal Health and Production.2019; 51(7): 1783. CrossRef
Seroprevalence of Neospora caninum infection in dairy cows in Northern provinces, Thailand
Tawin Inpankaew, Sathaporn Jittapalapong, Thomas Mitchell, Chainirun Sununta, Ikuo Igarashi, Xuenan Xuan
Acta Parasitologica.2014;[Epub] CrossRef
Risk factors of Neospora caninum infection in dogs and cats in dairy farms in Western Thailand
Pipat Arunvipas, Tawin Inpankaew, Sathaporn Jittapalapong
Tropical Animal Health and Production.2012; 44(5): 1117. CrossRef
Seroprevalence of Toxoplasma gondii and Neospora caninum in dogs from Korea
Thuy Nguyen, Se-Eun Choe, Jae-Won Byun, Hong-Bum Koh, Hee-Soo Lee, Seung-Won Kang
Acta Parasitologica.2012;[Epub] CrossRef
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D. Romero-Sal, Z. Garcia-Vaz, F. Montiel-Pa, T. Montiel-Pe, M. Aguilar-Do, L. Medina-Esp, C. Cruz-Vazqu
Journal of Animal and Veterinary Advances.2010; 9(10): 1445. CrossRef
Seroepidemiology of Neospora caninum and Toxoplasma gondii infection in yaks (Bos grunniens) in Qinghai, China
J. Liu, J.Z. Cai, W. Zhang, Q. Liu, D. Chen, J.P. Han, Q.R. Liu
Veterinary Parasitology.2008; 152(3-4): 330. CrossRef
Seroepidemiology of Neospora caninum and Toxoplasma gondii in cattle and water buffaloes (Bubalus bubalis) in the People's Republic of China
Jinhai Yu, Zhaofei Xia, Qun Liu, Jing Liu, Jun Ding, Wei Zhang
Veterinary Parasitology.2007; 143(1): 79. CrossRef
Epidemiology and Control of Neosporosis andNeospora caninum
J. P. Dubey, G. Schares, L. M. Ortega-Mora
Clinical Microbiology Reviews.2007; 20(2): 323. CrossRef
Diagnosis of bovine neosporosis
J.P. Dubey, G. Schares
Veterinary Parasitology.2006; 140(1-2): 1. CrossRef
ELISA detection of IgG antibody against a recombinant major surface antigen (Nc-p43) fragment of Neospora caninum in bovine sera
Hye-Jin Ahn, Sera Kim, Dae-Yong Kim, Ho-Woo Nam
The Korean Journal of Parasitology.2003; 41(3): 175. CrossRef
Seroprevalence of Neospora, Toxoplasma gondii and Sarcocystis neurona antibodies in horses from Jeju island, South Korea
G.D Gupta, J Lakritz, Jae-Hoon Kim, Dae-Yong Kim, Jin-Kap Kim, A.E Marsh
Veterinary Parasitology.2002; 106(3): 193. CrossRef
Determination of antigenic domain in GST fused major surface protein (Nc-p43) of Neospora caninum
Eui-Sun Son, Hye-Jin Ahn, Jae-Hoon Kim, Dae-Yong Kim, Ho-Woo Nam
The Korean Journal of Parasitology.2001; 39(3): 241. CrossRef

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Western blot analysis of stray cat sera against Toxoplasma gondii and the diagnostic availability of monoclonal antibodies in sandwich-ELISA: Woon-Mok Sohn, Ho-Woo Nam; Korean J Parasitol 1999;37(4):249-256.
Published online December 31, 1999; DOI: https://doi.org/10.3347/kjp.1999.37.4.249

Abstract
PDF

A total of 198 sera from stray cats was assayed against Toxoplasma gondii antigen by western blot. Out of 198 sera assayed, 26 sera (13.1%) showed typical blot patterns against T. gondii. When spotted by ELISA absorbance and indirect latex agglutination test (ILAT) titer, all 26 cases were distributed over the cut-off value of ELISA whereas 24 cases (92.3%) were in the positive range of 1:32 or higher and 2 cases in negative range by ILAT. Among western blot negative 172 sera, 162 cases were negative in both ILAT and ELISA while 10 cases were reactive falsely such that three cases were ILAT positive with 1:32 titer and 9 cases were ELISA positive (2 cases overlapped). These 10 cases reacted peculiarly without typical binding pattern in Western blot. Sandwich-ELISA was performed with monoclonal antibodies (mAbs) of Tg563 (30 kDa, SAG1), Tg505 (22 kDa, SAG2), Tg605 (43 kDa, SAG3), Tg556 (28 kDa, GRA2), Tg737 (32 kDa, GRA6), Tg695 (66 kDa, ROP2), Tg786 (42 kDa, ROP6), and Tg621 (32 kDa, anonymous but cytosolic) clone, respectively. All western blot-positive cases were in the positive range and negative cases in the negative range clearly. Among the 10 false reactive cases, 3 cases were in the positive range with one or more mAbs. All mAbs used in this study were confirmed to be specific to T. gondii infection as a standardized sandwich-ELISA to differentiate it from other pathogens.

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