||5R03CA077125-02 Interpret this number
||Cancer Prevention Instit Of California
||Reliability of EBV Detection in Hodgkins Disease Tumors
DESCRIPTION (Applicant's Description) This proposed R03 study will be the
first to examine observer reliability of EBER in situ hybridization (ISH)
and LMP-1 immunohistochemistry (IHC), morphologic molecular assays widely
used in clinical studies to detect Epstein-Barr virus (EBV) latent infection
of tumor cells. As these assays are easy to use, rapid, and applicable to
archived tumor specimens, they are ideally suited for large-scale molecular
epidemiologic projects. In Hodgkin's disease (HD), a common cancer of young
adults, EBER ISH and LMP-1 IHC have detected EBV in about 40 percent of
cases, and marked epidemiologic differences exist between the virus-defined
subgroups, However, EBV prevalence varies among HD studies even after
statistical control for epidemiologic differences in populations. Problems
with observer reliability may be partly responsible; yet, although assay
interpretation is subjective, reliability has never been evaluated for
either test. This project will determine inter- and intra-rater reliability
of EBER ISH and LMP-1 IHC assays in EBV detection in 40 HD tumors, based on
two reviews by each of four hematopathologists of slides, prepared at each
of two labs. From tumor specimens evaluated for prior projects, we will
randomly select 10 EBV-positive and 10 EBV-negative cases of each of the two
most common histology subtypes, to maximize precision of the kappa
statistic. From these 40 banked specimens, each lab will stain 40 slides
for EBER ISH and 40 for LMP-1 IHC, yielding 80 slides per assay. At NCCC,
stained slides from the two labs will be intermixed within assays, relabeled
to blind raters to institutional identification, and mailed in batches to
raters, who will record their interpretations (EBV-positive, EBV-negative)
and both rank and describe interpretive difficulty. Been lst and 2nd
reviews, staff will renumber slides to blind raters to the review sequence.
640 ratings (320 first and 320 second) will occur for each assay. With
these data, we will compute kappa statistics for multiple raters to
determine inter- and intra-rater agreement occurring beyond chance for each
assay. Kappas will also be compared for agreement on slides prepared on the
same specimens by each lab to detect any lab effect on interpretation
accuracy, and for the two histologic subtypes to detect differences that may
lead to variation in EBV prevalence by subtype. Findings will be developed
into specific interpretation guidelines. Study strengths include: the
first examination of both inter- and intra-rater agreement for two commonly
used clinical EBV detection assays with potential for epidemiologic
application in studying EBV-linked cancer; development of explicit
interpretation guidelines; use of existing pathology materials for
efficiency and for assembling a case series which maximizes precision;
adequate sample size to estimate inter-histology differences with sufficient
statistical power; collaboration with experts in the field of EBV detection;
and focus on HD, a common EBV-related cancer with both epidemiologic and
clinical importance for EBV classification.
Guidelines for interpreting EBER in situ hybridization and LMP1 immunohistochemical tests for detecting Epstein-Barr virus in Hodgkin lymphoma.
, Glaser S.L.
, Craig F.E.
, Borowitz M.
, Mann R.B.
, Shema S.J.
, Ambinder R.F.
American journal of clinical pathology, 2002 Feb; 117(2), p. 259-67.