BACKGROUND: The dramatic increase in use of the IgG test for toxoplasma, rubella, cytomegalovirus (CMV), and herpes simplex virus (HSV) [TORCH] has led to the requirement for a high-efficiency method that can be used in the clinical laboratory. This study aimed to compare the results of BGI-Array ELISA TORCH IgG (BGI-GBI, China) screening method to those of Virion/Serion TORCH IgG ELISA (Virion/Serion, Germany). METHODS: Serum specimens (n=400) submitted for routine IgG testing by Virion/Serion ELISA were also tested using the BGI-Array ELISA method. The agreements of these two kinds of method were analyzed by kappa-coefficients calculation. RESULTS: Following repeat testing, the BGI-Array ELISA TORCH IgG assays demonstrated agreements of 99.5% (398/400 specimens), 98% (392/400 specimens), 99% (396/400 specimens), and 99.5% (398/400 specimens), respectively. The BGI-Array ELISA IgG assays provided results comparable to Virion/Serion ELISA results, with kappa-coefficients showing near-perfect agreement for the HSV (kappa=0.87), rubella (kappa=0.92) and CMV (kappa=0.93) and substantial agreement for the toxoplasma (kappa=0.80) IgG assays. The use of the BGI-Array ELISA TORCH IgG assays could reduce the turnaround time (1.5 hr vs. 5 hr by Virion/Serion ELISA for 100 specimens) and were easy to use. CONCLUSIONS: BGI-Array ELISA TORCH IgG shows a good agreement with Virion/Serion ELISA methods and is suitable for clinical application.
Antibodies, Viral/blood ; Cytomegalovirus/immunology/*metabolism ; *Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin G/*analysis/blood ; Protozoan Infections/diagnosis ; Reagent Kits, Diagnostic ; Rubella virus/immunology/*metabolism ; Sensitivity and Specificity ; Simplexvirus/immunology/*metabolism ; Toxoplasma/immunology/*metabolism ; Virion/*immunology/metabolism ; Virus Diseases/diagnosis

OBJECTIVETo express the rubella virus E1-374 glycoprotein in Pichia pastoris and study the immunogenecity of the recombinant protein.

METHODSThe cDNA of protein E1-374 was cloned into the expression vector pGAPZalphaA and transformed into Pichia pastoris GS115 cells by electrotransfection. The expressed protein was confirmed by indirect immunofluorescence and demonstrated immunoreactivity by Western Blot. Rubella virus IgG antibody was assayed with ELISA after mice were inmmunized by E1-374 glycoprotein.

RESULTSSDS-PAGE analysis and Western Blot analysis of E1-374 protein revealed this protein to be 46.89 x 10(3). Antiserum (1:100) and E1-374 (5.5 microg/ml) was chosen for ELISA optimization. The intra-assay coefficient of variation for the ELISA was 0.36%-12.45%.

CONCLUSIONProtein E1-374 was highly expressed in Pichia pastoris cells, and it was a good choice to prepare rubella virus recombinant protein vaccines.

Animals ; Enzyme-Linked Immunosorbent Assay ; Female ; Gene Expression ; Humans ; Mice ; Mice, Inbred BALB C ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; genetics ; immunology ; Rubella virus ; genetics ; immunology ; Viral Envelope Proteins ; genetics ; immunology

OBJECTIVETo apply recombinant rubella virus envelope protein-1 (E1) to detect human rubella virus IgG antibody.

METHODSRubella virus E1 protein was expressed in E. coli, purified E1 protein was used as the antigen for the detecting of anti rubella in human sera in the way of enzyme linked Immunosorbant assay (ELISA).

RESULTSThe antigenicity of the recombinant protein was checked by WHO rubella sera panel. We detected 200 sera samples, which came from Guangxi Guilin. 93% of these samples were positive.

CONCLUSIONThe antigenicity of recombinant E1 is a satisfied candidate antigen for the detecting of human rubella virus antibody. The prevalence of anti rubella virus IgG in Guangxi is 93%. It is at the some level compared with other provinces in China.

Animals ; Antibodies, Viral ; analysis ; immunology ; Cercopithecus aethiops ; China ; Cloning, Molecular ; Escherichia coli ; genetics ; Gene Expression ; Genetic Vectors ; Humans ; Recombinant Proteins ; genetics ; immunology ; Rubella virus ; genetics ; immunology ; Vero Cells ; virology ; Viral Envelope Proteins ; genetics ; metabolism