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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 tests for serum IgG antibody responses to B. burgdorferi C6, B. garinii C6, and B. afzelii C6 in samples from EM patients.(A) Total sensitivity of PHOSPHAN variants G1, G2, G3, and G1–3 at the baseline prior to treatment (N = 146). (B) Sensitivity of PHOSPHAN variants G1–G3 in tests of 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: G1, Bb C6 IgG; G2, Bg C6 IgG; G3, Ba C6 IgG; G1–3, any of the three antigens Bb C6, Bg C6, or Ba C6. (Bb) B.burgdorferi, (Bg) B.garinii, (Ba) B.afzelii. Legend: (B) Transparent square, G1; Black diamond, G2; Black triangle, G3.
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pone.0130048.g004: Sensitivity of PHOSPHAN tests for serum IgG antibody responses to B. burgdorferi C6, B. garinii C6, and B. afzelii C6 in samples from EM patients.(A) Total sensitivity of PHOSPHAN variants G1, G2, G3, and G1–3 at the baseline prior to treatment (N = 146). (B) Sensitivity of PHOSPHAN variants G1–G3 in tests of 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: G1, Bb C6 IgG; G2, Bg C6 IgG; G3, Ba C6 IgG; G1–3, any of the three antigens Bb C6, Bg C6, or Ba C6. (Bb) B.burgdorferi, (Bg) B.garinii, (Ba) B.afzelii. Legend: (B) Transparent square, G1; Black diamond, G2; Black triangle, G3.

Mentions: In general, positive reactions for specific IgG with Bb C6 (G1), Bg C6 (G2), or Ba C6 (G3) were detected in 30.8–40.4% serum samples from LB patients at the baseline prior to treatment. The frequency of IgG detection with any of the three antigens (G1–3) was significantly higher only as compared to G2 (Fig 4A). In the course of the disease, the frequency of detection of IgG to Bb C6 increased from 40.4% at the baseline (95% CI: 32.4, 48.8) to a peak of 85.4% on days 15–30 after disease onset (95% CI: 72.2, 93.9) and then began to decrease. Parameters observed with Ba C6 were similar, while those with Bg C6 were markedly lower. Differences in IgG detection frequency between Bg C6 and two other C6 peptides were statistically significant only on days 15–30 from onset of illness (Fig 4B).


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 tests for serum IgG antibody responses to B. burgdorferi C6, B. garinii C6, and B. afzelii C6 in samples from EM patients.(A) Total sensitivity of PHOSPHAN variants G1, G2, G3, and G1–3 at the baseline prior to treatment (N = 146). (B) Sensitivity of PHOSPHAN variants G1–G3 in tests of 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: G1, Bb C6 IgG; G2, Bg C6 IgG; G3, Ba C6 IgG; G1–3, any of the three antigens Bb C6, Bg C6, or Ba C6. (Bb) B.burgdorferi, (Bg) B.garinii, (Ba) B.afzelii. Legend: (B) Transparent square, G1; Black diamond, G2; Black triangle, G3.
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Related In: Results  -  Collection

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pone.0130048.g004: Sensitivity of PHOSPHAN tests for serum IgG antibody responses to B. burgdorferi C6, B. garinii C6, and B. afzelii C6 in samples from EM patients.(A) Total sensitivity of PHOSPHAN variants G1, G2, G3, and G1–3 at the baseline prior to treatment (N = 146). (B) Sensitivity of PHOSPHAN variants G1–G3 in tests of 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: G1, Bb C6 IgG; G2, Bg C6 IgG; G3, Ba C6 IgG; G1–3, any of the three antigens Bb C6, Bg C6, or Ba C6. (Bb) B.burgdorferi, (Bg) B.garinii, (Ba) B.afzelii. Legend: (B) Transparent square, G1; Black diamond, G2; Black triangle, G3.
Mentions: In general, positive reactions for specific IgG with Bb C6 (G1), Bg C6 (G2), or Ba C6 (G3) were detected in 30.8–40.4% serum samples from LB patients at the baseline prior to treatment. The frequency of IgG detection with any of the three antigens (G1–3) was significantly higher only as compared to G2 (Fig 4A). In the course of the disease, the frequency of detection of IgG to Bb C6 increased from 40.4% at the baseline (95% CI: 32.4, 48.8) to a peak of 85.4% on days 15–30 after disease onset (95% CI: 72.2, 93.9) and then began to decrease. Parameters observed with Ba C6 were similar, while those with Bg C6 were markedly lower. Differences in IgG detection frequency between Bg C6 and two other C6 peptides were statistically significant only on days 15–30 from onset of illness (Fig 4B).

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