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"immunofluorescence"

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"immunofluorescence"

Original Article

Cryptosporidium Oocyst Detection in Water Samples: Floatation Technique Enhanced with Immunofluorescence Is as Effective as Immunomagnetic Separation Method
Khuanchai Koompapong, Chantira Sutthikornchai, Yowalark Sukthana
Korean J Parasitol 2009;47(4):353-357.
Published online December 1, 2009
DOI: https://doi.org/10.3347/kjp.2009.47.4.353

Cryptosporidium can cause gastrointestinal diseases worldwide, consequently posing public health problems and economic burden. Effective techniques for detecting contaminated oocysts in water are important to prevent and control the contamination. Immunomagnetic separation (IMS) method has been widely employed recently due to its efficiency, but, it is costly. Sucrose floatation technique is generally used for separating organisms by using their different specific gravity. It is effective and cheap but time consuming as well as requiring highly skilled personnel. Water turbidity and parasite load in water sample are additional factors affecting to the recovery rate of those 2 methods. We compared the efficiency of IMS and sucrose floatation methods to recover the spiked Cryptosporidium oocysts in various turbidity water samples. Cryptosporidium oocysts concentration at 1, 101, 102, and 103 per 10 ?l were spiked into 3 sets of 10 ml-water turbidity (5, 50, and 500 NTU). The recovery rate of the 2 methods was not different. Oocyst load at the concentration < 102 per 10 ml yielded unreliable results. Water turbidity at 500 NTU decreased the recovery rate of both techniques. The combination of sucrose floatation and immunofluorescense assay techniques (SF-FA) showed higher recovery rate than IMS and immunofluorescense assay (IMS-FA). We used this SF-FA to detect Cryptosporidium and Giardia from the river water samples and found 9 and 19 out of 30 (30% and 63.3%) positive, respectively. Our results favored sucrose floatation technique enhanced with immunofluorescense assay for detecting contaminated protozoa in water samples in general laboratories and in the real practical setting.

Citations

Citations to this article as recorded by  Crossref logo
  • Treatment of esophagogastric intussusception due to cryptosporidiosis using paromomycin in a corn snake (Pantherophis guttatus)
    Urarako SHIMIZU, Manabu KURIHARA, Kenichi TAMUKAI, Kazutaka YAMADA
    Journal of Veterinary Medical Science.2025; 87(4): 389.     CrossRef
  • Improved recovery of Cryptosporidium and Giardia in water by optimizing immunomagnetic separation using acid and heat dissociation
    Eun-Sook Lee, Ji-Yeon Lee, Young-Ae Baek, Seok-Ju Cho
    Journal of Water and Health.2025; 23(6): 757.     CrossRef
  • A smartphone microscopic method for simultaneous detection of (oo)cysts of Cryptosporidium and Giardia
    Retina Shrestha, Rojina Duwal, Sajeev Wagle, Samiksha Pokhrel, Basant Giri, Bhanu Bhakta Neupane, Joseph Mathu Ndung'u
    PLOS Neglected Tropical Diseases.2020; 14(9): e0008560.     CrossRef
  • Implications of new legislation (US FSMA) and guidelines (EC) on the establishment of management systems for agricultural water
    A. Allende, A.R. Datta, W.A. Smith, R. Adonis, A. MacKay, A.D. Adell
    Food Microbiology.2018; 75: 119.     CrossRef
  • Cryptosporidium and Giardia in Wastewater and Surface Water Environments
    Kerry A. Hamilton, Monique Waso, Brandon Reyneke, Nazanin Saeidi, Audrey Levine, Cindy Lalancette, Marie‐Claude Besner, Wesaal Khan, Warish Ahmed
    Journal of Environmental Quality.2018; 47(5): 1006.     CrossRef
  • Evolution of monitoring for Giardia and Cryptosporidium in water
    Artemis Efstratiou, Jerry Ongerth, Panagiotis Karanis
    Water Research.2017; 123: 96.     CrossRef
  • Report of fatal mixed infection with Cryptosporidium parvum and Giardia intestinalis in neonatal calves
    Yuu Matsuura, Makoto Matsubayashi, Satoko Nukata, Tomoyuki Shibahara, Osamu Ayukawa, Yasuko Kondo, Tomohide Matsuo, Shigehiko Uni, Masaru Furuya, Hiroyuki Tani, Naotoshi Tsuji, Kazumi Sasai
    Acta Parasitologica.2017;[Epub]     CrossRef
  • Rapid concentration and sensitive detection of hookworm ova from wastewater matrices using a real-time PCR method
    P. Gyawali, J.P.S. Sidhu, W. Ahmed, P. Jagals, S. Toze
    Experimental Parasitology.2015; 159: 5.     CrossRef
  • Prevalence and distribution of Cryptosporidium and Giardia in wastewater and the surface, drinking and ground waters in the Lower Rhine, Germany
    C. GALLAS-LINDEMANN, I. SOTIRIADOU, J. PLUTZER, P. KARANIS
    Epidemiology and Infection.2013; 141(1): 9.     CrossRef
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Brief Communications
Evaluation of α-Tubulin as an Antigenic and Molecular Probe to Detect Giardia lamblia
Juri Kim, Myeong Heon Shin, Kyoung-Ju Song, Soon-Jung Park
Korean J Parasitol 2009;47(3):287-291.
Published online August 28, 2009
DOI: https://doi.org/10.3347/kjp.2009.47.3.287

The α/β-tubulin heterodimer is the basic subunit of microtubules in eukaryotes. Polyclonal antibodies specific to recombinant α-tubulin of Giardia lamblia were made, and found effective as a probe to specifically detect G. lamblia by immunofluorescence assays. Nucleotide sequences of α-tubulin genes were compared between G. lamblia WB and GS strains, prototypes of assemblage A and assemblage B, respectively. A set of primers was designed and used to amplify a portion of the α-tubulin gene from G. lamblia. PCR-RFLP analysis of this α-tubulin PCR product successfully differentiated G. lamblia into 2 distinct groups, assemblages A and B. The results indicate that α-tubulin can be used as a molecular probe to detect G. lamblia.

Citations

Citations to this article as recorded by  Crossref logo
  • Genetic diversity and molecular diagnosis of Giardia
    Yankai Chang, Junqiang Li, Longxian Zhang
    Infection, Genetics and Evolution.2023; 113: 105482.     CrossRef
  • Tubulin as a potential molecular target for resveratrol in Giardia lamblia trophozoites, in vitro and in silico approaches
    José Roberto Vargas-Villanueva, Filiberto Gutiérrez-Gutiérrez, Mariana Garza-Ontiveros, Sendar Daniel Nery-Flores, Lizeth Guadalupe Campos-Múzquiz, Dagoberto Vazquez-Obregón, Raul Rodriguez-Herrera, Lissethe Palomo-Ligas
    Acta Tropica.2023; 248: 107026.     CrossRef
  • Phosphorylation of Serine 148 in Giardia lamblia End‐binding 1 Protein is Important for Cell Division
    Juri Kim, Hye‐Yeon Lee, Kyu‐Ho Lee, Soon‐Jung Park
    Journal of Eukaryotic Microbiology.2017; 64(4): 464.     CrossRef
  • Host defences against Giardia lamblia
    G. Lopez‐Romero, J. Quintero, H. Astiazarán‐García, C. Velazquez
    Parasite Immunology.2015; 37(8): 394.     CrossRef
  • Characterization of Microtubule-Binding and Dimerization Activity of Giardia lamblia End-Binding 1 Protein
    Juri Kim, Sara Nagami, Kyu-Ho Lee, Soon-Jung Park, Ira J. Blader
    PLoS ONE.2014; 9(5): e97850.     CrossRef
  • Zoonotic potential of Giardia
    Una Ryan, Simone M. Cacciò
    International Journal for Parasitology.2013; 43(12-13): 943.     CrossRef
  • Identification of Antigenic Proteins in Trichomonas vaginalis
    Hye-Yeon Lee, Sujin Hyung, Jong Woong Lee, Juri Kim, Myeong Heon Shin, Jae-Sook Ryu, Soon-Jung Park
    The Korean Journal of Parasitology.2011; 49(1): 79.     CrossRef
  • 8,402 View
  • 115 Download
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Immunolocalization of the 150 kDa protein in cyst fluid of Taenia solium metacestodes
Hyun-Jong Yang, Young-Bae Chung
Korean J Parasitol 2004;42(2):81-84.
Published online June 20, 2004
DOI: https://doi.org/10.3347/kjp.2004.42.2.81

The 150 kDa protein of cyst fluid (CF) of Taenia solium metacestodes was purified by ammonium sulfate fractionation and Superose 6 HR gel filtration chromatography. The purified protein consisted of three subunits (15, 10 and 7 kDa proteins), which were analyzed with the use of a 7.5-15% gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Immunofluorescence study was carried out by using immunize specific polyclonal antibody. Positive reactions were noticed at bladder walls, calcareous corpuscles, granules of cyst fluid and some host tissue surrounding the bladder wall of the metacestodes. These results suggest that the 150 kDa protein was secreted into host tissues, inducing immune responses in the host, and it may play important roles in the cellular physiology of the parasites.

Citations

Citations to this article as recorded by  Crossref logo
  • Taenia saginata: Production and characterization of monoclonal antibodies against Taenia saginata metacestode antigens
    Josy Campanhã Vicentini-Oliveira, Marjorie A. Golim, Silvana de Cássia Paulan, Germano Francisco Biondi, Rosana Rossi-Ferreira, Elenice Deffune, Cáris Maroni Nunes
    Experimental Parasitology.2010; 126(4): 621.     CrossRef
  • 7,902 View
  • 69 Download
  • Crossref