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Highlights on novel technologies for the detection of antibodies to Ro60, Ro52, and SS-B.

Infantino M, Bentow C, Seaman A, Benucci M, Atzeni F, Sarzi-Puttini P, Olivito B, Meacci F, Manfredi M, Mahler M - Clin. Dev. Immunol. (2013)

Bottom Line: There were 34 discrepant samples among all assays (20 anti-SS-B, 11 anti-Ro52, 3 anti-Ro60). 30/33 of retested samples (by D-tek dot blot) agreed with the QUANTA Flash results.QUANTA Flash and BioPlex 2200 show good qualitative agreement.The clinical performances were similar for anti-Ro52 and anti-Ro60 autoantibodies while differences were observed for anti-SS-B (La) antibodies.

View Article: PubMed Central - PubMed

Affiliation: Immunology and Allergology Laboratory Unit, Azienda Sanitaria di Firenze, San Giovanni di Dio Hospital, Florence, Italy.

ABSTRACT

Objective: We aimed to compare a chemiluminescent immunoassay (CIA, QUANTA Flash) on BIO-FLASH with a multiplex flow immunoassay (MFI) on BioPlex 2200 for the detection of antibodies to Ro60, Ro52, and SS-B.

Methods: The study included 241 samples, from patients suffering from systemic autoimmune diseases (n = 108) as well as disease controls (n = 133). All samples were tested for anti-Ro52, anti-Ro60, and anti-SS-B (La) antibodies on QUANTA Flash (INOVA Diagnostics, San Diego, USA) and BioPlex 2200 (Bio-Rad Laboratories Inc., Hercules, USA). Discrepant samples were tested by two independent methods: BlueDot/ANA and QUANTRIX Microarray (both D-tek, Belgium).

Results: The overall qualitative agreements were 95.4% (95% confidence interval, CI 92.0-97.7%) for anti-Ro52, 98.8% (95% CI 96.4-99.7%) for anti-Ro60, and 91.7% (95% CI 87.5-94.9%) for anti-SS-B antibodies. There were 34 discrepant samples among all assays (20 anti-SS-B, 11 anti-Ro52, 3 anti-Ro60). 30/33 of retested samples (by D-tek dot blot) agreed with the QUANTA Flash results. Similar findings were obtained with QUANTRIX Microarray kit.

Conclusion: QUANTA Flash and BioPlex 2200 show good qualitative agreement. The clinical performances were similar for anti-Ro52 and anti-Ro60 autoantibodies while differences were observed for anti-SS-B (La) antibodies.

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Related in: MedlinePlus

Supervised cluster analysis of the results. Supervised centered cluster analysis according to disease cohort is shown. The dendrogram shows that the Ro60 assays are closely related, the Ro52 assays are somewhat related, and the SS-B assays show significant difference. AT: atopic dermatitis; UCTD: undifferentiated connective tissue disease, RA: rheumatoid arthritis, SSc: systemic sclerosis; SED: suspected eye disease, SLE: systemic lupus erythematosus; HI: healthy individuals; SARD: systemic autoimmune rheumatic disease.
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fig4: Supervised cluster analysis of the results. Supervised centered cluster analysis according to disease cohort is shown. The dendrogram shows that the Ro60 assays are closely related, the Ro52 assays are somewhat related, and the SS-B assays show significant difference. AT: atopic dermatitis; UCTD: undifferentiated connective tissue disease, RA: rheumatoid arthritis, SSc: systemic sclerosis; SED: suspected eye disease, SLE: systemic lupus erythematosus; HI: healthy individuals; SARD: systemic autoimmune rheumatic disease.

Mentions: Discrepant samples (20 anti-SS-B, 11 anti-Ro52, 3 anti-Ro60) with sufficient residual volume (n = 33) were tested by two comparative methods. Thirty out of 33 discrepant samples among all assays (19 anti-SS-B, 11 anti-Ro52, 3 anti-Ro60) retested by D-tek dot blot agreed with the QUANTA Flash results (Table 3). Five of the 20 anti-SS-B discrepant samples were high positive on BioPlex (>8.0 units, above the AMR) and negative on QUANTA Flash, BlueDot, and QUANTRIX. Of the 17 BioPlex anti-SS-B positive/QUANTA Flash anti-SS-B negative samples 14 were from SARD and 4 from non-SARD patients (3 RA and one HI). All three QUANTA Flash anti-SS-B positive/BioPlex anti-SS-B negative samples were from SARD patients. All three were additionally anti-Ro60 antibody positive by both methods (QUANTA Flash and BioPlex). Figure 4 depicts a cluster analysis of all methods sorted by disease group, which serves as a visual aid of the antibody prevalence and differences between methods.


Highlights on novel technologies for the detection of antibodies to Ro60, Ro52, and SS-B.

Infantino M, Bentow C, Seaman A, Benucci M, Atzeni F, Sarzi-Puttini P, Olivito B, Meacci F, Manfredi M, Mahler M - Clin. Dev. Immunol. (2013)

Supervised cluster analysis of the results. Supervised centered cluster analysis according to disease cohort is shown. The dendrogram shows that the Ro60 assays are closely related, the Ro52 assays are somewhat related, and the SS-B assays show significant difference. AT: atopic dermatitis; UCTD: undifferentiated connective tissue disease, RA: rheumatoid arthritis, SSc: systemic sclerosis; SED: suspected eye disease, SLE: systemic lupus erythematosus; HI: healthy individuals; SARD: systemic autoimmune rheumatic disease.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3860079&req=5

fig4: Supervised cluster analysis of the results. Supervised centered cluster analysis according to disease cohort is shown. The dendrogram shows that the Ro60 assays are closely related, the Ro52 assays are somewhat related, and the SS-B assays show significant difference. AT: atopic dermatitis; UCTD: undifferentiated connective tissue disease, RA: rheumatoid arthritis, SSc: systemic sclerosis; SED: suspected eye disease, SLE: systemic lupus erythematosus; HI: healthy individuals; SARD: systemic autoimmune rheumatic disease.
Mentions: Discrepant samples (20 anti-SS-B, 11 anti-Ro52, 3 anti-Ro60) with sufficient residual volume (n = 33) were tested by two comparative methods. Thirty out of 33 discrepant samples among all assays (19 anti-SS-B, 11 anti-Ro52, 3 anti-Ro60) retested by D-tek dot blot agreed with the QUANTA Flash results (Table 3). Five of the 20 anti-SS-B discrepant samples were high positive on BioPlex (>8.0 units, above the AMR) and negative on QUANTA Flash, BlueDot, and QUANTRIX. Of the 17 BioPlex anti-SS-B positive/QUANTA Flash anti-SS-B negative samples 14 were from SARD and 4 from non-SARD patients (3 RA and one HI). All three QUANTA Flash anti-SS-B positive/BioPlex anti-SS-B negative samples were from SARD patients. All three were additionally anti-Ro60 antibody positive by both methods (QUANTA Flash and BioPlex). Figure 4 depicts a cluster analysis of all methods sorted by disease group, which serves as a visual aid of the antibody prevalence and differences between methods.

Bottom Line: There were 34 discrepant samples among all assays (20 anti-SS-B, 11 anti-Ro52, 3 anti-Ro60). 30/33 of retested samples (by D-tek dot blot) agreed with the QUANTA Flash results.QUANTA Flash and BioPlex 2200 show good qualitative agreement.The clinical performances were similar for anti-Ro52 and anti-Ro60 autoantibodies while differences were observed for anti-SS-B (La) antibodies.

View Article: PubMed Central - PubMed

Affiliation: Immunology and Allergology Laboratory Unit, Azienda Sanitaria di Firenze, San Giovanni di Dio Hospital, Florence, Italy.

ABSTRACT

Objective: We aimed to compare a chemiluminescent immunoassay (CIA, QUANTA Flash) on BIO-FLASH with a multiplex flow immunoassay (MFI) on BioPlex 2200 for the detection of antibodies to Ro60, Ro52, and SS-B.

Methods: The study included 241 samples, from patients suffering from systemic autoimmune diseases (n = 108) as well as disease controls (n = 133). All samples were tested for anti-Ro52, anti-Ro60, and anti-SS-B (La) antibodies on QUANTA Flash (INOVA Diagnostics, San Diego, USA) and BioPlex 2200 (Bio-Rad Laboratories Inc., Hercules, USA). Discrepant samples were tested by two independent methods: BlueDot/ANA and QUANTRIX Microarray (both D-tek, Belgium).

Results: The overall qualitative agreements were 95.4% (95% confidence interval, CI 92.0-97.7%) for anti-Ro52, 98.8% (95% CI 96.4-99.7%) for anti-Ro60, and 91.7% (95% CI 87.5-94.9%) for anti-SS-B antibodies. There were 34 discrepant samples among all assays (20 anti-SS-B, 11 anti-Ro52, 3 anti-Ro60). 30/33 of retested samples (by D-tek dot blot) agreed with the QUANTA Flash results. Similar findings were obtained with QUANTRIX Microarray kit.

Conclusion: QUANTA Flash and BioPlex 2200 show good qualitative agreement. The clinical performances were similar for anti-Ro52 and anti-Ro60 autoantibodies while differences were observed for anti-SS-B (La) antibodies.

Show MeSH
Related in: MedlinePlus