TY - JOUR
T1 - Standardization of fluorescence in situ hybridization studies on chronic lymphocytic leukemia (CLL) blood and marrow cells by the CLL Research Consortium
AU - Smoley, Stephanie A.
AU - Van Dyke, Daniel L.
AU - Kay, Neil E.
AU - Heerema, Nyla A.
AU - Dell' Aquila, Marie L.
AU - Dal Cin, Paola
AU - Koduru, Prasad
AU - Aviram, Ayala
AU - Rassenti, Laura
AU - Byrd, John C.
AU - Rai, Kanti R.
AU - Brown, Jennifer R.
AU - Greaves, Andrew W.
AU - Eckel-Passow, Jeanette
AU - Neuberg, Donna
AU - Kipps, Thomas J.
AU - Dewald, Gordon W.
N1 - Funding Information:
This work was partially supported National Insitutes of Health (NIH) grant PO1-CA81534 of the CRC and NIH grant CA95241 of PI Neil Kay. We acknowledge the CRC members and thank the staff of the CRC Cytogenetics Laboratories for their expert technical support.
PY - 2010/12
Y1 - 2010/12
N2 - Five laboratories in the Chronic Lymphocytic Leukemia (CLL) Research Consortium (CRC) investigated standardizing and pooling of fluorescence in situ hybridization (FISH) results as a collaborative research project. This investigation used fixed bone marrow and blood cells available from previous conventional cytogenetic or FISH studies in two pilot studies, a one-day workshop, and proficiency test. Multiple FISH probe strategies were used to detect 6q-, 11q-, +12, 13q-, 17p-, and IGH rearrangements. Ten specimens were studied by participants who used their own probes (pilot study 1). Of 312 FISH interpretations, 224 (72%) were true-negative, 74 (24%) true-positive, 6 (2%) false-negative, and 8 (3%) false-positive. In pilot study no. 2, each participant studied two specimens using identical FISH probe sets to control for variation due to probe sets and probe strategies. Of 80 FISH interpretations, no false interpretations were identified. At a subsequent workshop, discussions produced agreement on scoring criteria. The proficiency test that followed produced no false-negative results and 4% (3/68) false-positive interpretations. Interpretation disagreements among laboratories were primarily attributable to inadequate normal cutoffs, inconsistent scoring criteria, and the use of different FISH probe strategies. Collaborative organizations that use pooled FISH results may wish to impose more conservative empiric normal cutoff values or use an equivocal range between the normal cutoff and the abnormal reference range to eliminate false-positive interpretations. False-negative results will still occur, and would be expected in low-percentage positive cases; these would likely have less clinical significance than false positive results. Individual laboratories can help by closely following rigorous quality assurance guidelines to ensure accurate and consistent FISH studies in their clinical practice and research.
AB - Five laboratories in the Chronic Lymphocytic Leukemia (CLL) Research Consortium (CRC) investigated standardizing and pooling of fluorescence in situ hybridization (FISH) results as a collaborative research project. This investigation used fixed bone marrow and blood cells available from previous conventional cytogenetic or FISH studies in two pilot studies, a one-day workshop, and proficiency test. Multiple FISH probe strategies were used to detect 6q-, 11q-, +12, 13q-, 17p-, and IGH rearrangements. Ten specimens were studied by participants who used their own probes (pilot study 1). Of 312 FISH interpretations, 224 (72%) were true-negative, 74 (24%) true-positive, 6 (2%) false-negative, and 8 (3%) false-positive. In pilot study no. 2, each participant studied two specimens using identical FISH probe sets to control for variation due to probe sets and probe strategies. Of 80 FISH interpretations, no false interpretations were identified. At a subsequent workshop, discussions produced agreement on scoring criteria. The proficiency test that followed produced no false-negative results and 4% (3/68) false-positive interpretations. Interpretation disagreements among laboratories were primarily attributable to inadequate normal cutoffs, inconsistent scoring criteria, and the use of different FISH probe strategies. Collaborative organizations that use pooled FISH results may wish to impose more conservative empiric normal cutoff values or use an equivocal range between the normal cutoff and the abnormal reference range to eliminate false-positive interpretations. False-negative results will still occur, and would be expected in low-percentage positive cases; these would likely have less clinical significance than false positive results. Individual laboratories can help by closely following rigorous quality assurance guidelines to ensure accurate and consistent FISH studies in their clinical practice and research.
UR - http://www.scopus.com/inward/record.url?scp=78649999975&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=78649999975&partnerID=8YFLogxK
U2 - 10.1016/j.cancergencyto.2010.08.009
DO - 10.1016/j.cancergencyto.2010.08.009
M3 - Article
C2 - 21156226
AN - SCOPUS:78649999975
SN - 0165-4608
VL - 203
SP - 141
EP - 148
JO - Cancer Genetics and Cytogenetics
JF - Cancer Genetics and Cytogenetics
IS - 2
ER -