2015
DOI: 10.1371/journal.pone.0121647
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Exploring Early and Late Toxoplasma gondii Strain RH Infection by Two-Dimensional Immunoblots of Chicken Immunoglobulin G and M Profiles

Abstract: Toxoplasma gondii is an intracellular apicomplexan parasite infecting warm-blooded vertebrate hosts, with only early infection stage being contained with drugs. But diagnosis differencing early and late infection was not available. In the present investigation, 2-dimensional immunobloting was used to explore early and late infections in chickens. The protein expression of T. gondii was determined by image analysis of the tachyzoites proteome separated by standard-one and conventional two-dimentional gel polyac… Show more

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Cited by 5 publications
(6 citation statements)
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“…The climatic conditions, lighting program, and chicken fodder and water were manually-operated and the chicks were cared for in agreement with the approved guidelines of the Institutional Animal Care and Committee of China Agricultural University. Sporulated oocysts (2.5 × 10 3 ) were inoculated into broiler chicks by oral gavage, and faeces were collected at 5–9 days post-inoculation according to eimerian species and the experimental requirements 19 20 21 . The collected oocysts of Eimeria mitis were used to spike other oocyst-free chicken faeces.…”
Section: Methodsmentioning
confidence: 99%
“…The climatic conditions, lighting program, and chicken fodder and water were manually-operated and the chicks were cared for in agreement with the approved guidelines of the Institutional Animal Care and Committee of China Agricultural University. Sporulated oocysts (2.5 × 10 3 ) were inoculated into broiler chicks by oral gavage, and faeces were collected at 5–9 days post-inoculation according to eimerian species and the experimental requirements 19 20 21 . The collected oocysts of Eimeria mitis were used to spike other oocyst-free chicken faeces.…”
Section: Methodsmentioning
confidence: 99%
“…The protein preparation was performed according to [ 10 ]. Protein was prepared by rapid freezing and thawing 3 times using liquid nitrogen to disrupt sporozoites in lysis buffer containing 7 M urea, 2 M Thiourea, 4%(w/v) CHAPS, 1%(w/v) DTT, 1 mM PMSF and 0.5%(v/v) IPG buffer pH (3–10) dissolved in 40 mM Tris-base pH 9.6.…”
Section: Methodsmentioning
confidence: 99%
“…Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) of sporozoite proteins was conducted according to the procedure of [ 10 ] as follows: Sporozoite antigens of two E . maxima strains were subjected to nonlinear immobilized pH gradient strips (IPG) (Immobiline DryStrip, pH 3–10 nonlinear, 13cm, Amersham, Uppsala, Sweden) using the Ettan IPGphor Isoelectric Focusing system (Amersham).…”
Section: Methodsmentioning
confidence: 99%
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“…Toxoplasma gondii is an obligate intracellular protozoan parasite, which can infect almost all warmblooded, including humans, leading to toxoplasmosis [1,2,3,4,5]. Approximately 30% of the world's population has serological evidence of Toxoplasma infection [6].…”
Section: Introductionmentioning
confidence: 99%