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Production and Evaluation of Toxoplasma gondii Recombinant Sur-face Antigen 1 (SAG1) for Serodiagnosis of Acute and ChronicToxop-lasma Infection in Human Sera
M (Mina) Selseleh,H Keshavarz,M Mohebali,S Shojaee
Iranian Journal of Parasitology , 2012,
Abstract: Background: The assays currently available for the detection of specific anti-Toxoplasma antibodies may vary in their abilities to detect serum immunoglobulins, due to the Lack of a purified standardized antigen. The aim of this study was evaluation the recombinant Toxoplasma gondii SAG1 antigen for the serodiagnosis of acute and chronic toxoplasmosis. Methods: This study describes an ELISA using recombinant SAG1 for detection of IgM and IgG antibodies against Toxoplasma gondii in human sera. Genomic DNA of T. gondii (RH Strain) was isolated and PCR reaction was performed. Recovered DNA was cloned into PTZ57R cloning vector. The recombinant plasmid was detected by restriction analysis. The SAG1 gene was subcloned in the pET- 28a expression vector. Protein production was then induced with 1 mM isopropyl-D - thiogalactopyrano-side (IPTG). A total of 204 sera were tested using a commercial IgG and IgM ELISA kit (Trinity, USA) as gold standard prior to testing them with the recombinant antigen. Results: Tested sera were divided into the following groups:(a) The 74 T. gondii IgG positive (b) 70 T.gondii IgM positive (c) 60 sera who had no serological evidence of toxoplasmosis as negative sera.To determine the specificity of the test, we used other parasitic diseases including echinococusis (N=5), malaria (N=14), leishmania-sis (N=7),fasciolasis (N=4 ), sterengyloidiasis (N=1 ). Sensitivity and specificity of the generated recombinant IgG ELISA in comparison with commercial ELISA (Com ELISA) were 93% and 95%, and the sensitivity and specificity of the generated recombinant IgM ELISA were 87% and 95% respectively. Conclusion: The results acquired here show that this antigen is useful for diagnostic purposes and could be replaced by lysed, whole cell antigens for diagnosis of chronic toxoplasmosis.
A Case Report of Blood Group Discrepancy because of Anti A1 with Clinical Significance and Anti C
Esmaeili, J.,Ebrahimy, P.,Selseleh, M.,Babadivand, P
Medical Laboratory Journal , 2012, DOI: http://www.goums.ac.ir/mljgoums/index.php?&slct_pg_id=10&sid=1&slc_lang=en
Abstract: Background and objectives: ABO phenotyping is one of theessential tests in Immunohematology. Incompatible blood grouptransfusion leads to acute hemolysis reactions and other seriouscomplications. Anti A1 is a cold Antibody with no clinicalsignificance, but if it is reacted at 37 c can be clinically significant,which will be happened rarely. At the present, we report a Case withanti A1 having clinical significance and Anti C.Material and methods: The patient was suffering from Paroxysmalnocturnal hemoglobinuria (PNH) and received repeated bloodtransfusion. The tests performed for this patient were blood grouping,Antibody screening, panel test and cross match.Result: the patient’s blood group is AB (A2B) with anti A1. Thepresence of anti C is verified by applying Panel test.Conclusion: to prepare appropriate blood, Cross mach test wasperformed on A2B blood bags without C Antigen. During laboratorytests and blood transfusion, no reaction was observed. This reportindicates that being aware of anti-antigen antibodies is an importantpoint.Key words: Anti C and Anti A1 with Clinical importance, PNH, AbScreenin
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