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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.


Box plots (minimum, first quartile, median value, third quartile, and maximum value) of median fluorescence intensity (MFI) values from single antigen bead assays (SAB) that yielded inconsistent results among the five participating laboratories. Results are grouped by the different MFI cutoff values (1,000, 3,000, 5,000, and 10,000); upper panel contains data for HLA class I antibody detection; lower panel contains data for HLA class II antibody detection.*P<0.05; **P<0.01 by Mann-Whitney test.
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Figure 2: Box plots (minimum, first quartile, median value, third quartile, and maximum value) of median fluorescence intensity (MFI) values from single antigen bead assays (SAB) that yielded inconsistent results among the five participating laboratories. Results are grouped by the different MFI cutoff values (1,000, 3,000, 5,000, and 10,000); upper panel contains data for HLA class I antibody detection; lower panel contains data for HLA class II antibody detection.*P<0.05; **P<0.01 by Mann-Whitney test.

Mentions: The median MFI values for the results that were discordant among the five laboratories at each cutoff value are depicted in Fig. 2. The MFI values for the Class I beads from the SN and BH laboratories were lower than those from the YH, CH, and SH research groups at the 1,000, 3,000, and 5,000 MFI cutoffs (all P<0.01). Additionally, at the 10,000 MFI cutoff, the values from the BH and SH groups were significantly lower than those from SN, YH, and CH (all P<0.01). Meanwhile, the MFI values for the Class II beads from the CH laboratory were significantly higher than those from SN, BH, and SH groups at the 1,000 and 3,000 MFI cutoffs (all P<0.01), and the MFI values from the SN group were significantly lower than those from BH, CH, and SH at the 10,000 MFI cutoff (all P<0.01).


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)

Box plots (minimum, first quartile, median value, third quartile, and maximum value) of median fluorescence intensity (MFI) values from single antigen bead assays (SAB) that yielded inconsistent results among the five participating laboratories. Results are grouped by the different MFI cutoff values (1,000, 3,000, 5,000, and 10,000); upper panel contains data for HLA class I antibody detection; lower panel contains data for HLA class II antibody detection.*P<0.05; **P<0.01 by Mann-Whitney test.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 2: Box plots (minimum, first quartile, median value, third quartile, and maximum value) of median fluorescence intensity (MFI) values from single antigen bead assays (SAB) that yielded inconsistent results among the five participating laboratories. Results are grouped by the different MFI cutoff values (1,000, 3,000, 5,000, and 10,000); upper panel contains data for HLA class I antibody detection; lower panel contains data for HLA class II antibody detection.*P<0.05; **P<0.01 by Mann-Whitney test.
Mentions: The median MFI values for the results that were discordant among the five laboratories at each cutoff value are depicted in Fig. 2. The MFI values for the Class I beads from the SN and BH laboratories were lower than those from the YH, CH, and SH research groups at the 1,000, 3,000, and 5,000 MFI cutoffs (all P<0.01). Additionally, at the 10,000 MFI cutoff, the values from the BH and SH groups were significantly lower than those from SN, YH, and CH (all P<0.01). Meanwhile, the MFI values for the Class II beads from the CH laboratory were significantly higher than those from SN, BH, and SH groups at the 1,000 and 3,000 MFI cutoffs (all P<0.01), and the MFI values from the SN group were significantly lower than those from BH, CH, and SH at the 10,000 MFI cutoff (all P<0.01).

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.