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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

Lyme Index values (LIV) for sera from LB patients with erythema migrans (N = 351) and healthy blood donors (N = 197).(A) PHOSPHAN variants M1–M3 and (B) PHOSPHAN variants G1–G3 and C6 ELISA (C6). Blood donors, *; line with arrowhead, threshold level (LIV = 1). Immunoassay codes: M1, Bb C6 IgM; M2, OspC IgM; M3, VlsE IgM; G1, Bb C6 IgG; G2, Bg C6 IgG; G3, Ba C6 IgG; C6, Immunetics C6 Lyme ELISA kit. (Bb) B.burgdorferi, (Bg) B.garinii, (Ba) B.afzelii. Legend: Transparent square, Mean; Spotted rectangle, ± Standard Error; Dotted rectangle, Non-Outlier Range; Transparent circle, Outliers; Black square, Extremes.
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pone.0130048.g001: Lyme Index values (LIV) for sera from LB patients with erythema migrans (N = 351) and healthy blood donors (N = 197).(A) PHOSPHAN variants M1–M3 and (B) PHOSPHAN variants G1–G3 and C6 ELISA (C6). Blood donors, *; line with arrowhead, threshold level (LIV = 1). Immunoassay codes: M1, Bb C6 IgM; M2, OspC IgM; M3, VlsE IgM; G1, Bb C6 IgG; G2, Bg C6 IgG; G3, Ba C6 IgG; C6, Immunetics C6 Lyme ELISA kit. (Bb) B.burgdorferi, (Bg) B.garinii, (Ba) B.afzelii. Legend: Transparent square, Mean; Spotted rectangle, ± Standard Error; Dotted rectangle, Non-Outlier Range; Transparent circle, Outliers; Black square, Extremes.

Mentions: Fig 1 illustrates the results of IgM and IgG detection with each of six antigens by PHOSPHAN and of IgM/IgG detection by C6 ELISA. Both methods allowed differentiation between LB patients and healthy donors. In PHOSPHAN tests, the average LI values for IgM detection with antigens Bb C6 (M1), OspC (M2), or VlsE (M3) in the healthy donors were in the range of 0.27–0.34, compared to 1.28–1.97 in the LB group (Fig 1A). The respective values for IgG detection with antigens Bb C6 (G1), Bg C6 (G2), or Ba C6 (G3) were 0.17–0.37 and 20.6–26.2; in C6 ELISA, they were 0.54 (95% CI: 0.12, 2.1) and 3.4 (95% CI: 0.2, 7.2) (Fig 1B). Compared to the control samples, the average LI values for the LB group were higher by factors of 5.6 (IgM detection) and 70 (IgG detection) in PHOSPHAN tests and by a factor of 6.4 in the C6 ELISA. The highest LI values for serum samples from the LB group ranged from 18–29 in PHOSPHAN variants M1, M2, or M3 (Fig 1A) and from 287–462 in variants G1, G2, or G3; the highest values recorded in the C6 ELISA did not exceed 12.3 (Fig 1B). Coefficients of variation in the LI values obtained after repeated analysis of the same samples on different days were slightly higher in PHOSPHAN than in the C6 ELISA (up to 25% vs. no more than 15%).


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)

Lyme Index values (LIV) for sera from LB patients with erythema migrans (N = 351) and healthy blood donors (N = 197).(A) PHOSPHAN variants M1–M3 and (B) PHOSPHAN variants G1–G3 and C6 ELISA (C6). Blood donors, *; line with arrowhead, threshold level (LIV = 1). Immunoassay codes: M1, Bb C6 IgM; M2, OspC IgM; M3, VlsE IgM; G1, Bb C6 IgG; G2, Bg C6 IgG; G3, Ba C6 IgG; C6, Immunetics C6 Lyme ELISA kit. (Bb) B.burgdorferi, (Bg) B.garinii, (Ba) B.afzelii. Legend: Transparent square, Mean; Spotted rectangle, ± Standard Error; Dotted rectangle, Non-Outlier Range; Transparent circle, Outliers; Black square, Extremes.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4492927&req=5

pone.0130048.g001: Lyme Index values (LIV) for sera from LB patients with erythema migrans (N = 351) and healthy blood donors (N = 197).(A) PHOSPHAN variants M1–M3 and (B) PHOSPHAN variants G1–G3 and C6 ELISA (C6). Blood donors, *; line with arrowhead, threshold level (LIV = 1). Immunoassay codes: M1, Bb C6 IgM; M2, OspC IgM; M3, VlsE IgM; G1, Bb C6 IgG; G2, Bg C6 IgG; G3, Ba C6 IgG; C6, Immunetics C6 Lyme ELISA kit. (Bb) B.burgdorferi, (Bg) B.garinii, (Ba) B.afzelii. Legend: Transparent square, Mean; Spotted rectangle, ± Standard Error; Dotted rectangle, Non-Outlier Range; Transparent circle, Outliers; Black square, Extremes.
Mentions: Fig 1 illustrates the results of IgM and IgG detection with each of six antigens by PHOSPHAN and of IgM/IgG detection by C6 ELISA. Both methods allowed differentiation between LB patients and healthy donors. In PHOSPHAN tests, the average LI values for IgM detection with antigens Bb C6 (M1), OspC (M2), or VlsE (M3) in the healthy donors were in the range of 0.27–0.34, compared to 1.28–1.97 in the LB group (Fig 1A). The respective values for IgG detection with antigens Bb C6 (G1), Bg C6 (G2), or Ba C6 (G3) were 0.17–0.37 and 20.6–26.2; in C6 ELISA, they were 0.54 (95% CI: 0.12, 2.1) and 3.4 (95% CI: 0.2, 7.2) (Fig 1B). Compared to the control samples, the average LI values for the LB group were higher by factors of 5.6 (IgM detection) and 70 (IgG detection) in PHOSPHAN tests and by a factor of 6.4 in the C6 ELISA. The highest LI values for serum samples from the LB group ranged from 18–29 in PHOSPHAN variants M1, M2, or M3 (Fig 1A) and from 287–462 in variants G1, G2, or G3; the highest values recorded in the C6 ELISA did not exceed 12.3 (Fig 1B). Coefficients of variation in the LI values obtained after repeated analysis of the same samples on different days were slightly higher in PHOSPHAN than in the C6 ELISA (up to 25% vs. no more than 15%).

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