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C6 Peptide-Based Multiplex Phosphorescence Analysis (PHOSPHAN) for Serologic Confirmation of Lyme Borreliosis.

Pomelova VG, Korenberg EI, Kuznetsova TI, Bychenkova TA, Bekman NI, Osin NS - PLoS ONE (2015)

Bottom Line: 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.IgM 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%).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.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Diagnostics, Department of Biological Microassay, State Research Institute of Biological Engineering, Moscow, Russian Federation.

ABSTRACT

Background: A 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.

Objective: To evaluate the performance of C6 peptide based multiplex Phosphorescence Analysis (PHOSPHAN) for the serologic confirmation of Lyme borreliosis (LB) in Russian patients.

Methods: Serum 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.

Results: IgM 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.

Conclusions: The 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.

No MeSH data available.


Related in: MedlinePlus

Sensitivity of PHOSPHAN and C6 ELISA tests for serum IgM and IgG antibody responses to B. burgdorferi C6 in samples from EM patients.(A) Sensitivity of PHOSPHAN variants M1, G1, M1G1 and C6 ELISA (C6) at the baseline prior to treatment (N = 146). (B) Sensitivity of PHOSPHAN variants M1 and G1, and (C) PHOSPHAN variant M1G1 and C6 ELISA in tests of serum samples taken at the baseline (n = 146) and on days 8–14 (n = 75), 15–30 (n = 48), and 31–66 (n = 82) after disease onset. Values that are statistically significant (Fisher's exact test, p < 0.05) are denoted by brackets or asterisks. Immunoassay codes: M1, Bb C6 IgM; G1, Bb C6 IgG; M1G1, Bb C6 IgM+IgG; C6, Immunetics C6 Lyme ELISA kit. (Bb) B.burgdorferi. Legend: (B) Transparent square, M1; Black diamond, G1. (C) Black square, M1G1; Transparent circle, C6.
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pone.0130048.g002: Sensitivity of PHOSPHAN and C6 ELISA tests for serum IgM and IgG antibody responses to B. burgdorferi C6 in samples from EM patients.(A) Sensitivity of PHOSPHAN variants M1, G1, M1G1 and C6 ELISA (C6) at the baseline prior to treatment (N = 146). (B) Sensitivity of PHOSPHAN variants M1 and G1, and (C) PHOSPHAN variant M1G1 and C6 ELISA in tests of serum samples taken at the baseline (n = 146) and on days 8–14 (n = 75), 15–30 (n = 48), and 31–66 (n = 82) after disease onset. Values that are statistically significant (Fisher's exact test, p < 0.05) are denoted by brackets or asterisks. Immunoassay codes: M1, Bb C6 IgM; G1, Bb C6 IgG; M1G1, Bb C6 IgM+IgG; C6, Immunetics C6 Lyme ELISA kit. (Bb) B.burgdorferi. Legend: (B) Transparent square, M1; Black diamond, G1. (C) Black square, M1G1; Transparent circle, C6.

Mentions: In general, a positive PHOSPHAN reaction with C6 from B. burgdorferi was observed significantly more frequently in tests for IgG (G1) than for IgM (M1) both at the baseline prior to treatment and at all time intervals postbaseline (Fig 2A and 2B). The frequency of positive samples in tests for both IgM and IgG (M1G1) was higher than in variant G1 (Fig 2A), although the difference lacked statistical significance (p = 0.07). The sensitivity of C6 ELISA was comparable to that of the PHOSPHAN variant for IgG detection (G1), but it was significantly lower than the total IgM and IgG variant assay (M1G1) (Fig 2A). Difference in sensitivity between PHOSPHAN and C6 ELISA was especially apparent at the baseline (p < 0.05), whereas no statistically significant difference was observed at the later time intervals after start of treatment (Fig 2C).


C6 Peptide-Based Multiplex Phosphorescence Analysis (PHOSPHAN) for Serologic Confirmation of Lyme Borreliosis.

Pomelova VG, Korenberg EI, Kuznetsova TI, Bychenkova TA, Bekman NI, Osin NS - PLoS ONE (2015)

Sensitivity of PHOSPHAN and C6 ELISA tests for serum IgM and IgG antibody responses to B. burgdorferi C6 in samples from EM patients.(A) Sensitivity of PHOSPHAN variants M1, G1, M1G1 and C6 ELISA (C6) at the baseline prior to treatment (N = 146). (B) Sensitivity of PHOSPHAN variants M1 and G1, and (C) PHOSPHAN variant M1G1 and C6 ELISA in tests of serum samples taken at the baseline (n = 146) and on days 8–14 (n = 75), 15–30 (n = 48), and 31–66 (n = 82) after disease onset. Values that are statistically significant (Fisher's exact test, p < 0.05) are denoted by brackets or asterisks. Immunoassay codes: M1, Bb C6 IgM; G1, Bb C6 IgG; M1G1, Bb C6 IgM+IgG; C6, Immunetics C6 Lyme ELISA kit. (Bb) B.burgdorferi. Legend: (B) Transparent square, M1; Black diamond, G1. (C) Black square, M1G1; Transparent circle, C6.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4492927&req=5

pone.0130048.g002: Sensitivity of PHOSPHAN and C6 ELISA tests for serum IgM and IgG antibody responses to B. burgdorferi C6 in samples from EM patients.(A) Sensitivity of PHOSPHAN variants M1, G1, M1G1 and C6 ELISA (C6) at the baseline prior to treatment (N = 146). (B) Sensitivity of PHOSPHAN variants M1 and G1, and (C) PHOSPHAN variant M1G1 and C6 ELISA in tests of serum samples taken at the baseline (n = 146) and on days 8–14 (n = 75), 15–30 (n = 48), and 31–66 (n = 82) after disease onset. Values that are statistically significant (Fisher's exact test, p < 0.05) are denoted by brackets or asterisks. Immunoassay codes: M1, Bb C6 IgM; G1, Bb C6 IgG; M1G1, Bb C6 IgM+IgG; C6, Immunetics C6 Lyme ELISA kit. (Bb) B.burgdorferi. Legend: (B) Transparent square, M1; Black diamond, G1. (C) Black square, M1G1; Transparent circle, C6.
Mentions: In general, a positive PHOSPHAN reaction with C6 from B. burgdorferi was observed significantly more frequently in tests for IgG (G1) than for IgM (M1) both at the baseline prior to treatment and at all time intervals postbaseline (Fig 2A and 2B). The frequency of positive samples in tests for both IgM and IgG (M1G1) was higher than in variant G1 (Fig 2A), although the difference lacked statistical significance (p = 0.07). The sensitivity of C6 ELISA was comparable to that of the PHOSPHAN variant for IgG detection (G1), but it was significantly lower than the total IgM and IgG variant assay (M1G1) (Fig 2A). Difference in sensitivity between PHOSPHAN and C6 ELISA was especially apparent at the baseline (p < 0.05), whereas no statistically significant difference was observed at the later time intervals after start of treatment (Fig 2C).

Bottom Line: 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.IgM 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%).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.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Diagnostics, Department of Biological Microassay, State Research Institute of Biological Engineering, Moscow, Russian Federation.

ABSTRACT

Background: A 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.

Objective: To evaluate the performance of C6 peptide based multiplex Phosphorescence Analysis (PHOSPHAN) for the serologic confirmation of Lyme borreliosis (LB) in Russian patients.

Methods: Serum 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.

Results: IgM 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.

Conclusions: The 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.

No MeSH data available.


Related in: MedlinePlus