Test ID: SYPGR    
Syphilis IgG Antibody with Reflex, Serum

The BioPlex Syphilis IgG is a multiplex flow immunoassay performed on the BioPlex 2200 System (Bio-Rad Laboratories, Hercules, CA). Three types of dyed beads are coated with recombinant proteins associated with Treponema pallidum (15kDa, 17kDa and 47kDa). An aliquot of patient sample, sample diluent, and bead reagent are combined in a reaction vessel, and the mixture is incubated at 37 degrees C. After a wash cycle, antihuman IgG antibody conjugated to phycoerythrin (PE) is added to the dyed beads, and this mixture is incubated at 37 degrees C. Excess conjugate is removed in another wash cycle and beads are resuspended in wash buffer. The bead mixture is then passed through a flow-based detector and identified according to the fluorescence emitted, which is specific to the internal dye composition of the microsphere. The amount of antibody bound to the capture antigen is then determined by measuring the fluorescence of the attached PE. Raw data is calculated in relative fluorescence intensity (RFI).(Package insert: BioPlex 2200 System Syphilis IgG [T.pallidum], Bio-Rad Laboratories Clinical Diagnostic Group, Hercules, CA)

If the IgG result is positive, a rapid plasma regain (RPR) is automatically performed. The RPR test is a macroscopic screening assay done with unheated serum. Reagin reacts with nontreponemal antigen containing colloidal charcoal particles. This reaction results in a visual flocculation of the black particles against the white card background. The test yields a positive or negative result, and all positive samples are titered to determine the highest positive dilution.(Huber TW, Storms S, Young P, et al: Reactivity of microhemagglutination, fluorescent treponemal antibody absorption, Venereal Disease Research Laboratory, and rapid plasma reagin tests in primary syphilis. J Clin Microbiol 1983 Mar;17[3]:405-409)

The Serodia TP-PA test is based on the agglutination of colored gelatin particle carriers sensitized with Treponema pallidum (Nichols Strain) antigen. Serum samples are serially diluted in microplate wells. Sensitized gelatin particles are added to respective wells and the contents of the plate mixed.  The mixture is incubated for 2 hours at room temperature. Serum containing specific antibodies will react with the antigen-sensitized colored gelatin particles to form a smooth mat of agglutinated particles in the microplate well. A compact button formed by the settling of the non-agglutinated particles characterizes negative reactions. The agglutination patterns are read visually to determine interpretation.(Package insert: Serodia -TP PA, Fujirebio Diagnostics, Inc., Tokyo, Japan.)

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