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Interlaboratory Comparison of the Results of Lifecodes LSA Class I and Class II Single Antigen Kits for Human Leukocyte Antigen Antibody Detection.

Oh EJ, Park H, Park KU, Kang ES, Kim HS, Song EY - Ann Lab Med (2015)

Bottom Line: The categorical results obtained from the five laboratories exhibited concordance rates of 96.0% and 97.2% for detection of HLA class I and class II antibodies, respectively.The median CVs for the MFI values among five laboratories in the lower MFI range (<1,000) were significantly higher than those for the other MFI ranges (all P<0.01).Analysis of SAB performed in five laboratories using identical protocols and reagents from the same lot resulted in high levels of concordance and strong correlation of results.

View Article: PubMed Central - PubMed

Affiliation: Department of Laboratory Medicine, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul, Korea.

ABSTRACT

Background: Although single antigen bead assays (SAB) are approved qualitative tests, the median fluorescence intensity (MFI) values obtained from SAB are frequently used in combination with quantitative significances for diagnostic purposes. To gauge the reproducibility of SAB results, we assessed the interlaboratory variability of MFI values using identical kits with reagents from the same lot and the manufacturer's protocol.

Methods: Six serum samples containing HLA-specific antibodies were analyzed at five laboratories by using Lifecodes LSA Class I and Class II SAB kits (Immucor, USA) from the same lot, according to the manufacturer's protocol. We analyzed the concordance of qualitative results according to distinct MFI cutoffs (1,000, 3,000, 5,000, and 10,000), and the correlation of quantitative MFI values obtained by the participating laboratories. The CV for MFI values were analyzed and grouped by mean MFI values from the five laboratories (<1,000; 1,000-2,999; 3,000-4,999; 5,000-9,999; and ≥10,000).

Results: The categorical results obtained from the five laboratories exhibited concordance rates of 96.0% and 97.2% for detection of HLA class I and class II antibodies, respectively. The Pearson correlation coefficients for MFI values of class I and class II antibodies were between 0.947-0.991 and 0.992-0.997, respectively. The median CVs for the MFI values among five laboratories in the lower MFI range (<1,000) were significantly higher than those for the other MFI ranges (all P<0.01).

Conclusions: Analysis of SAB performed in five laboratories using identical protocols and reagents from the same lot resulted in high levels of concordance and strong correlation of results.

No MeSH data available.


Concordance of single antigen bead assay (SAB) results among the five participating laboratories. SAB analysis was performed by using six serum samples and four different median fluorescence intensity (MFI) cutoffs. The X-axis shows the different MFI cutoffs, and the Y-axis shows the frequency of concordant results. (A) Class I; (B) Class II beads.
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Figure 1: Concordance of single antigen bead assay (SAB) results among the five participating laboratories. SAB analysis was performed by using six serum samples and four different median fluorescence intensity (MFI) cutoffs. The X-axis shows the different MFI cutoffs, and the Y-axis shows the frequency of concordant results. (A) Class I; (B) Class II beads.

Mentions: The concordance of results was assessed for each bead. Results for a particular bead were considered concordant only if the participating five laboratories obtained the same outcome (either positive or negative). For class I antigens, 94.8-97.3% of the beads yielded concordant results in the six sera among the laboratories at four different MFI cutoffs: 1,000, 3,000, 5,000, and 10,000 (Fig. 1A). When an MFI of 1,000 was used as the cutoff, 95.5% of the 558 beads (93 beads×6 sera) yielded concordant results; 68.6% of the beads were consistently negative, and 26.9% of beads were consistently positive. At MFI cutoffs of 3,000, 5,000, and 10,000, 94.8% (529/558 beads, 79.2% negative and 15.6% positive), 97.3% (543/558 beads, 85.7% negative and 11.6% positive) and 96.4% (538/558 beads, 93.4% negative and 3.0% positive) of the beads exhibited concordant results, respectively. Overall, 96.0% (2,143/2,232 beads) of the beads yielded consistent results in the participating laboratories for detection of HLA class I antibodies.


Interlaboratory Comparison of the Results of Lifecodes LSA Class I and Class II Single Antigen Kits for Human Leukocyte Antigen Antibody Detection.

Oh EJ, Park H, Park KU, Kang ES, Kim HS, Song EY - Ann Lab Med (2015)

Concordance of single antigen bead assay (SAB) results among the five participating laboratories. SAB analysis was performed by using six serum samples and four different median fluorescence intensity (MFI) cutoffs. The X-axis shows the different MFI cutoffs, and the Y-axis shows the frequency of concordant results. (A) Class I; (B) Class II beads.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4390700&req=5

Figure 1: Concordance of single antigen bead assay (SAB) results among the five participating laboratories. SAB analysis was performed by using six serum samples and four different median fluorescence intensity (MFI) cutoffs. The X-axis shows the different MFI cutoffs, and the Y-axis shows the frequency of concordant results. (A) Class I; (B) Class II beads.
Mentions: The concordance of results was assessed for each bead. Results for a particular bead were considered concordant only if the participating five laboratories obtained the same outcome (either positive or negative). For class I antigens, 94.8-97.3% of the beads yielded concordant results in the six sera among the laboratories at four different MFI cutoffs: 1,000, 3,000, 5,000, and 10,000 (Fig. 1A). When an MFI of 1,000 was used as the cutoff, 95.5% of the 558 beads (93 beads×6 sera) yielded concordant results; 68.6% of the beads were consistently negative, and 26.9% of beads were consistently positive. At MFI cutoffs of 3,000, 5,000, and 10,000, 94.8% (529/558 beads, 79.2% negative and 15.6% positive), 97.3% (543/558 beads, 85.7% negative and 11.6% positive) and 96.4% (538/558 beads, 93.4% negative and 3.0% positive) of the beads exhibited concordant results, respectively. Overall, 96.0% (2,143/2,232 beads) of the beads yielded consistent results in the participating laboratories for detection of HLA class I antibodies.

Bottom Line: The categorical results obtained from the five laboratories exhibited concordance rates of 96.0% and 97.2% for detection of HLA class I and class II antibodies, respectively.The median CVs for the MFI values among five laboratories in the lower MFI range (<1,000) were significantly higher than those for the other MFI ranges (all P<0.01).Analysis of SAB performed in five laboratories using identical protocols and reagents from the same lot resulted in high levels of concordance and strong correlation of results.

View Article: PubMed Central - PubMed

Affiliation: Department of Laboratory Medicine, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul, Korea.

ABSTRACT

Background: Although single antigen bead assays (SAB) are approved qualitative tests, the median fluorescence intensity (MFI) values obtained from SAB are frequently used in combination with quantitative significances for diagnostic purposes. To gauge the reproducibility of SAB results, we assessed the interlaboratory variability of MFI values using identical kits with reagents from the same lot and the manufacturer's protocol.

Methods: Six serum samples containing HLA-specific antibodies were analyzed at five laboratories by using Lifecodes LSA Class I and Class II SAB kits (Immucor, USA) from the same lot, according to the manufacturer's protocol. We analyzed the concordance of qualitative results according to distinct MFI cutoffs (1,000, 3,000, 5,000, and 10,000), and the correlation of quantitative MFI values obtained by the participating laboratories. The CV for MFI values were analyzed and grouped by mean MFI values from the five laboratories (<1,000; 1,000-2,999; 3,000-4,999; 5,000-9,999; and ≥10,000).

Results: The categorical results obtained from the five laboratories exhibited concordance rates of 96.0% and 97.2% for detection of HLA class I and class II antibodies, respectively. The Pearson correlation coefficients for MFI values of class I and class II antibodies were between 0.947-0.991 and 0.992-0.997, respectively. The median CVs for the MFI values among five laboratories in the lower MFI range (<1,000) were significantly higher than those for the other MFI ranges (all P<0.01).

Conclusions: Analysis of SAB performed in five laboratories using identical protocols and reagents from the same lot resulted in high levels of concordance and strong correlation of results.

No MeSH data available.