Evaluation of α-Tubulin as an Antigenic and Molecular Probe to Detect Giardia lamblia

Kim, Juri; Shin, Myoung-Ho; Song, Kyoung-Ju; Park, Soon‐Jung

doi:10.3347/kjp.2009.47.3.287

Cited by 9 publications

(6 citation statements)

References 24 publications

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“…In the present work, were detected new proteins for specific IgA antibody response against G. intestinalis antigens in the fractions with reactivity for proteins of 205, 176, 110kDa, a region between 76 to 41kDa and the band of 23kDa which were mainly recognized in the fraction 2. The protein of 41kDa could belong to α or β-tubulin, proteins present in the cytoskeleton and the flagella on the parasite M [25]. Other proteins have also been identified by proteomics studies similar to those detected in the present work such as the proteins of 65 to 80kDa [26,27].…”

Section: Discussionsupporting

confidence: 80%

Parasite-specific IgA response in infected children by Giardia intestinalis from rural communities of Venezuela.

Durán¹,

Jiménez²,

Pocaterra³

et al. 2018

J Parasit Dis Diagn Ther

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Giardiasis is an infection caused by Giardia intestinalis into the small intestine that affects children in school age. The aim of the work was evaluated the specific IgA anti-Giardia antibodies in serum of children from rural communities were evaluated 100 children of 3 to 13 years old and measurement the levels of IgA antibody response against immunodominant antigens of Giardia intestinalis by ELISA and Westernblot. The soluble extract of Giardia intestinalis and fractions were obtained by differential centrifugation. Electrophoretic pattern showed several proteins from 205 to 14kDa, and sera of infected children recognized proteins of 205, 176, a region from 76 to 41kDa, and a band of 23kDa in the fraction 2. The IgA antibody response anti-Giardia for to soluble extract was higher in infected children compared to uninfected children (0.29 ± 0.16 vs. 0.07 ± 0.04; p ≤ 0.05). The IgA antibody response against the fraction 2 was higher in infected group than controls (1.0 ± 0.2 vs. 0.75 ± 0.26; p ≥ 0.05), the fraction 3 showed similar response to the soluble extract (0.45 ± 0.12 vs. 0.3 ± 0.12; p ≥ 0.05). The infection by Giardia intestinalis produce specific IgA antibodies directed to the proteins of the fraction 2. These proteins could be tested as antigens for serological diagnosis of giardiasis.

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Section: Discussionsupporting

confidence: 80%

Parasite-specific IgA response in infected children by Giardia intestinalis from rural communities of Venezuela.

Durán¹,

Jiménez²,

Pocaterra³

et al. 2018

J Parasit Dis Diagn Ther

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“…[613] The parasite is found to be the most prevalent intestinal protozoan[14] and is taken into WHO's Neglected Diseases Initiative recently. [15] Its direct and easy life cycle, robust cysts that resist water chlorination and environmental conditions, contamination of food, water, and fomites; and its zoonotic potential, make it a common human parasite[26] The prevalence of human giardiasis in Iran is reported to be 10.9% and it is the most prevalent intestinal parasite in this country.…”

Section: Discussionmentioning

confidence: 99%

Identification of genotypes of Giardia duodenalis human isolates in Isfahan, Iran, using polymerase chain reaction - Restriction Fragment Length polymorphism

et al. 2012

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Background:Giardia duodenalis is one of the most prevalent intestinal parasites of human. It also infects a wide range of mammals. Two genotype of G.duodenalis (A and B) were commonly reported among humans with different frequency of distribution in different geographical locations. This work was conducted to discriminate genotypes of Giardia duodenalis human isolates in Isfahan city using PCR- RFLP. This is the first molecular study on human isolates of G.duodenalis in the area.Methods:Samples were collected from different health centers of Isfahan city during June 2011 and February 2012. From 175 Giardia positive stool samples 67 specimens were selected randomly. Cysts of Giardia positive samples were concentrated by flotation sucrose. Extraction of genomic DNA from trophozoite and cysts was performed using QIAamp Stool Mini kit with a modified protocol. PCR- RFLP method was used to amplify a fragment of 458bp at the glutamate dehydrogenase locus, and restriction enzymes BspLI and RsaI differentiated human genotypes A and B and their subgroups.Results:PCR – RFLP assay of 67 isolates showed 40(59.7%) isolates as Genotype A group II, 23(34.32%) samples as Genotype B Group III and two (2.98%) sample as Genotype B group IV. Mixed genotype of (AII and B) was detected only in two isolates (2.98%).Conclusions:PCR – RFLP assay targeting gdh locus is a sensitive tool and discriminates genotypes, sub genotypes and mixed type of G.duodenalis. Results of our study suggest both anthroponotic and zoonotic origins for the infections respectively.

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“…Following incubation with horseradish peroxidase (HRP)-conjugated secondary antibodies, the immunoreactive protein was visualized using an enhanced chemiluminescence (ECL) system (Amersham Pharmacia). Membranes were incubated in a stripping buffer (Thermo Scientific) at room temperature for 30 min, and then reacted with polyclonal rat antibodies specific to the α-tubulin of G. lamblia (1∶10000) [21] .…”

Section: Methodsmentioning

confidence: 99%

Characterization of Microtubule-Binding and Dimerization Activity of Giardia lamblia End-Binding 1 Protein

et al. 2014

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End-binding 1 (EB1) proteins are evolutionarily conserved components of microtubule (MT) plus-end tracking protein that regulate MT dynamics. Giardia lamblia, with two nuclei and cytoskeletal structures, requires accurate MT distribution for division. In this study, we show that a single EB1 homolog gene of G. lamblia regulates MT dynamics in mitosis. The haemagglutinin-tagged G. lamblia EB1 (GlEB1) localizes to the nuclear envelopes and median bodies, and is transiently present in mitotic spindles of dividing cells. Knockdown of GlEB1 expression using the morpholinos-based anti-EB1 oligonucleotides, resulted in a significant defect in mitosis of Giardia trophozoites. The MT-binding assays using recombinant GlEB1 (rGlEB1) proteins demonstrated that rGlEB1102–238, but not rGlEB11–184, maintains an MT-binding ability comparable with that of the full length protein, rGlEB11–238. Size exclusion chromatography showed that rGlEB1 is present as a dimer formed by its C-terminal domain and a disulfide bond. In vitro-mutagenesis of GlEB1 indicated that an intermolecular disulfide bond is made between cysteine #13 of the two monomers. Complementation assay using the BIM1 knockout mutant yeast, the yeast homolog of mammalian EB1, indicated that expression of the C13S mutant GlEB1 protein cannot rescue the mitotic defect of the BIM1 mutant yeast. These results suggest that dimerization of GlEB1 via the 13th cysteine residues plays a role during mitosis in Giardia.

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Evaluation of α-Tubulin as an Antigenic and Molecular Probe to Detect Giardia lamblia

Cited by 9 publications

References 24 publications

Parasite-specific IgA response in infected children by Giardia intestinalis from rural communities of Venezuela.

Parasite-specific IgA response in infected children by Giardia intestinalis from rural communities of Venezuela.

Identification of genotypes of Giardia duodenalis human isolates in Isfahan, Iran, using polymerase chain reaction - Restriction Fragment Length polymorphism

Characterization of Microtubule-Binding and Dimerization Activity of Giardia lamblia End-Binding 1 Protein

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