Inter and intra laboratory concordance of HLA antibody results obtained by single antigen bead based assay

Manish J. Gandhi, Steven DeGoey, Deborah Falbo, Sarah Jenkins, James R. Stubbs, Harriet Noreen, David F. Lorentzen, JarHow Lee, Mark D Stegall

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Single antigen bead based assays (SAB) to identify antibodies to HLA are marketed as a qualitative test, however often used as a quantitative test as the results provide mean fluorescence intensity (MFI) which is found to correlate with the strength/avidity of the antibody. We studied the between and within laboratory variability in performing the SAB from one manufacturer. Ten samples were tested at four laboratories according to the manufacturer's suggested protocol. Additionally same samples were tested on four consecutive days in one laboratory. All tests were performed using the same lot of beads and secondary antibody. Results were classified as positive at four different MFI cutoffs: 1000, 5000, 8000 and 10,000. MFI values across and within the laboratory were compared in a pair-wise fashion with Pearson's correlations. Overall concordance for Class-I was 97% between laboratories and 98% within laboratory at all cutoffs. Pair-wise Pearson correlation between laboratories was 0.989-0.99, while within laboratory it was 0.998-0.999. For Class-II, overall concordance between and within laboratory was 98%. Pair-wise Pearson correlation between laboratories was 0.991-0.997, while within laboratory it was 0.997-0.999. There is good correlation between laboratories and within laboratory using the same manufacturer, same lot and same protocol while performing SAB.

Original languageEnglish (US)
Pages (from-to)310-317
Number of pages8
JournalHuman Immunology
Volume74
Issue number3
DOIs
StatePublished - Mar 2013

Fingerprint

Antigens
Antibodies
Fluorescence
Antibody Affinity

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

Cite this

Inter and intra laboratory concordance of HLA antibody results obtained by single antigen bead based assay. / Gandhi, Manish J.; DeGoey, Steven; Falbo, Deborah; Jenkins, Sarah; Stubbs, James R.; Noreen, Harriet; Lorentzen, David F.; Lee, JarHow; Stegall, Mark D.

In: Human Immunology, Vol. 74, No. 3, 03.2013, p. 310-317.

Research output: Contribution to journalArticle

Gandhi, MJ, DeGoey, S, Falbo, D, Jenkins, S, Stubbs, JR, Noreen, H, Lorentzen, DF, Lee, J & Stegall, MD 2013, 'Inter and intra laboratory concordance of HLA antibody results obtained by single antigen bead based assay', Human Immunology, vol. 74, no. 3, pp. 310-317. https://doi.org/10.1016/j.humimm.2012.12.003
Gandhi, Manish J. ; DeGoey, Steven ; Falbo, Deborah ; Jenkins, Sarah ; Stubbs, James R. ; Noreen, Harriet ; Lorentzen, David F. ; Lee, JarHow ; Stegall, Mark D. / Inter and intra laboratory concordance of HLA antibody results obtained by single antigen bead based assay. In: Human Immunology. 2013 ; Vol. 74, No. 3. pp. 310-317.
@article{36ac5c833da949b091c4eed2b8d78521,
title = "Inter and intra laboratory concordance of HLA antibody results obtained by single antigen bead based assay",
abstract = "Single antigen bead based assays (SAB) to identify antibodies to HLA are marketed as a qualitative test, however often used as a quantitative test as the results provide mean fluorescence intensity (MFI) which is found to correlate with the strength/avidity of the antibody. We studied the between and within laboratory variability in performing the SAB from one manufacturer. Ten samples were tested at four laboratories according to the manufacturer's suggested protocol. Additionally same samples were tested on four consecutive days in one laboratory. All tests were performed using the same lot of beads and secondary antibody. Results were classified as positive at four different MFI cutoffs: 1000, 5000, 8000 and 10,000. MFI values across and within the laboratory were compared in a pair-wise fashion with Pearson's correlations. Overall concordance for Class-I was 97{\%} between laboratories and 98{\%} within laboratory at all cutoffs. Pair-wise Pearson correlation between laboratories was 0.989-0.99, while within laboratory it was 0.998-0.999. For Class-II, overall concordance between and within laboratory was 98{\%}. Pair-wise Pearson correlation between laboratories was 0.991-0.997, while within laboratory it was 0.997-0.999. There is good correlation between laboratories and within laboratory using the same manufacturer, same lot and same protocol while performing SAB.",
author = "Gandhi, {Manish J.} and Steven DeGoey and Deborah Falbo and Sarah Jenkins and Stubbs, {James R.} and Harriet Noreen and Lorentzen, {David F.} and JarHow Lee and Stegall, {Mark D}",
year = "2013",
month = "3",
doi = "10.1016/j.humimm.2012.12.003",
language = "English (US)",
volume = "74",
pages = "310--317",
journal = "Human Immunology",
issn = "0198-8859",
publisher = "Elsevier Inc.",
number = "3",

}

TY - JOUR

T1 - Inter and intra laboratory concordance of HLA antibody results obtained by single antigen bead based assay

AU - Gandhi, Manish J.

AU - DeGoey, Steven

AU - Falbo, Deborah

AU - Jenkins, Sarah

AU - Stubbs, James R.

AU - Noreen, Harriet

AU - Lorentzen, David F.

AU - Lee, JarHow

AU - Stegall, Mark D

PY - 2013/3

Y1 - 2013/3

N2 - Single antigen bead based assays (SAB) to identify antibodies to HLA are marketed as a qualitative test, however often used as a quantitative test as the results provide mean fluorescence intensity (MFI) which is found to correlate with the strength/avidity of the antibody. We studied the between and within laboratory variability in performing the SAB from one manufacturer. Ten samples were tested at four laboratories according to the manufacturer's suggested protocol. Additionally same samples were tested on four consecutive days in one laboratory. All tests were performed using the same lot of beads and secondary antibody. Results were classified as positive at four different MFI cutoffs: 1000, 5000, 8000 and 10,000. MFI values across and within the laboratory were compared in a pair-wise fashion with Pearson's correlations. Overall concordance for Class-I was 97% between laboratories and 98% within laboratory at all cutoffs. Pair-wise Pearson correlation between laboratories was 0.989-0.99, while within laboratory it was 0.998-0.999. For Class-II, overall concordance between and within laboratory was 98%. Pair-wise Pearson correlation between laboratories was 0.991-0.997, while within laboratory it was 0.997-0.999. There is good correlation between laboratories and within laboratory using the same manufacturer, same lot and same protocol while performing SAB.

AB - Single antigen bead based assays (SAB) to identify antibodies to HLA are marketed as a qualitative test, however often used as a quantitative test as the results provide mean fluorescence intensity (MFI) which is found to correlate with the strength/avidity of the antibody. We studied the between and within laboratory variability in performing the SAB from one manufacturer. Ten samples were tested at four laboratories according to the manufacturer's suggested protocol. Additionally same samples were tested on four consecutive days in one laboratory. All tests were performed using the same lot of beads and secondary antibody. Results were classified as positive at four different MFI cutoffs: 1000, 5000, 8000 and 10,000. MFI values across and within the laboratory were compared in a pair-wise fashion with Pearson's correlations. Overall concordance for Class-I was 97% between laboratories and 98% within laboratory at all cutoffs. Pair-wise Pearson correlation between laboratories was 0.989-0.99, while within laboratory it was 0.998-0.999. For Class-II, overall concordance between and within laboratory was 98%. Pair-wise Pearson correlation between laboratories was 0.991-0.997, while within laboratory it was 0.997-0.999. There is good correlation between laboratories and within laboratory using the same manufacturer, same lot and same protocol while performing SAB.

UR - http://www.scopus.com/inward/record.url?scp=84873715740&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84873715740&partnerID=8YFLogxK

U2 - 10.1016/j.humimm.2012.12.003

DO - 10.1016/j.humimm.2012.12.003

M3 - Article

C2 - 23238217

AN - SCOPUS:84873715740

VL - 74

SP - 310

EP - 317

JO - Human Immunology

JF - Human Immunology

SN - 0198-8859

IS - 3

ER -