BackgroundA single-tier immunoassay using the C6 peptide of VlsE (C6) from Borrelia burgdorferi sensu stricto (Bb) has been proposed as a potential alternative to conventional two-tier testing for the serologic diagnosis of Lyme disease in the United States and Europe.ObjectiveTo evaluate the performance of C6 peptide based multiplex Phosphorescence Analysis (PHOSPHAN) for the serologic confirmation of Lyme borreliosis (LB) in Russian patients.MethodsSerum samples (n = 351) were collected from 146 patients with erythema migrans (EM); samples from 131 of these patients were taken several times prior to treatment and at different stages of recovery. The control group consisted of 197 healthy blood donors and 31 patients with other diseases, all from the same highly endemic region of Russia. All samples were analyzed by PHOSPHAN for IgM and IgG to Bb C6, recombinant OspC and VlsE proteins, and C6 peptides from B. garinii and B. afzelii.ResultsIgM and IgG to Bb C6 were identified in 43 and 95 out of 131 patients (32.8 and 72.5%, respectively); seroconversion of IgM antibodies was observed in about half of the patients (51.2%), and of IgG antibodies, in almost all of them (88.4%). Additional detection of OspC-IgM and VlsE-IgM or IgG to C6 from B. garinii or B. afzelii did not contribute significantly to the overall sensitivity of the multiplex immunoassay.ConclusionsThe multiplex phosphorescence immunoassay is a promising method for simultaneously revealing the spectrum of antibodies to several Borrelia antigens. Detection of IgM and IgG to Bb C6 in the sera of EM patients provides effective serologic confirmation of LB and, with high probability, indicates an active infection process.

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We have developed a multiplex immunoassay test (immunochip) based on PHOSPHAN technology for the detection of immunoglobulin G to herpes simplex virus (HSV) types 1, 2 and cytomegalovirus (CMV). The immunochip consists of HSV type specific gG1 (HSV-1) and gG2 (HSV-2) recombinant antigens, the lysate antigen for detection of total IgG to both HSV types (HSV 1/2), and CMV specific chimeric recombinant antigen containing the immunodominant sequences of pp150, gB, pp28 and pp52 proteins. The sensitivity and specificity of simultaneous IgGs detection with recombinant proteins were comparable to the commercial ELISA kits regardless of the kind of investigated serum specimens (patient sera, standard serum panels). The lysate HSV antigen was as sensitive but significantly less specific, so that it could not be recommended for use as a component of the multiplex test. These results can be used as a basis for creating commercial multiplex tests intended for high-productive screening of HSV, CMV and other TORCH-infections in a clinical laboratory.

We evaluated the utility of Borrelia burgdorferi sensu stricto (Bb) peptide C6 for serologic confirmation of Ixodid Tick-Borne Borrelioses (ITBB) in Russia. Serum samples (N = 1089) were from erythema migrans (EM) (N = 327) and the EM-free (EMF) patients (N = 115); in some patients, the disease was accompanied by human granulocytic anaplasmosis or tick-borne encephalitis. The sera were investigated by multiplex phosphorescence analysis (PHOSPHAN) for IgM to Bb C6, recombinant OspC and VlsE proteins, and IgG to C6 from Bb, B. garinii (Bg), and B. afzelii (Ba). Detection of Bb C6 IgM/IgG provided effective serologic confirmation of ITBB in both EM and EMF patients early after disease onset. In the EM-free patients, however, this test needed to be supplemented with detection of VlsE IgM in convalescent-phase sera due to delay in development of the antibody responses for C6 IgG. In general, positive PHOSPHAN reactions were observed in 81.9% and 86.7% of the EM and EMF patients, respectively, as well as in 59 of 65 (90.8%) patients, whose blood contained B. burgdorferi sensu lato DNA. Additional detection of IgG to Bg C6 or Ba C6 had no significant contribution to serologic diagnosis of ITBB in both patient groups.