Toward quality assurance for metaphase FISH: A multicenter experience

Gordon W. Dewald, Richard Stallard, Patricia I. Bader, Kathy Chen, Julie Zenger-Hain, Catherine J. Harris, Rodney Higgins, Betsy Hirsch, Wei Tong Hsu, Eric Johnson, Virginia Kubic, T. W. Kurczynski, James M. Malone, D. James McCorquodale, Karen Meilinger, Lorraine F. Meisner, J. W. Moore, Stuart Schwartz, Steven Siembieda, Patrick D. StortoGail Vance, Peter Van Tuinen, Ann Wiktor, Jar Fee Yung

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Although fluorescent in situ hybridization (FISH) is rapidly becoming a part of clinical cytogenetics, no organization sponsors multicenter determinations of the efficacy of probes. We report on 23 laboratories that volunteered to provide slides and to use a probe for small nuclear ribonucleoprotein polypeptide N (SNRPN) and a control locus. Experiences with FISH for these laboratories during 1994 ranged from 0 to 645 utilizations (median = 84) involving blood, amniotic fluid, and bone marrow. In an initial study of hybridization efficiency, the median percentage of metaphases from normal individuals showing two SNRPN and two control signals for slides prepared at each site was 97.0 (range = 74-100); for slides prepared by a central laboratory, it was 97.8 (range = 81.6-100). In a subsequent blind study, each laboratory attempted to score 5 metaphases from each of 23 specimens [8 with del(15)(q11.2→q12) and 15 with normal 15 chromosomes]. Of 529 challenges, the correct SNRPN pattern was found in 5 of 5 metaphases in 457 (86%) and in 4 of 5 in 33 (6%). Ambiguous, incomplete, or no results were reported for 32 (6%) challenges. Seven (1%) diagnostic errors were made, including 6 false positives and 1 false negative: 1 laboratory made 3 errors, 1 made 2, and 2 made 1 each. Most errors and inconsistencies seemed due to inexperience with FISH. The working time to process and analyze slides singly averaged 49.5 min; slides processed in batches of 4 and analyzed singly required 36.9 min. We conclude that proficiency testing for FISH by using an extensive array of challenges is possible and that multiple centers can collaborate to test probes and to evaluate costs.

Original languageEnglish (US)
Pages (from-to)190-196
Number of pages7
JournalAmerican Journal of Medical Genetics
Volume65
Issue number3
DOIs
StatePublished - Oct 28 1996

Fingerprint

Metaphase
Fluorescence In Situ Hybridization
Small Nuclear Ribonucleoproteins
Peptides
Chromosomes, Human, Pair 15
Internal-External Control
Amniotic Fluid
Diagnostic Errors
Cytogenetics
Bone Marrow
Costs and Cost Analysis

Keywords

  • hybridization efficiency
  • metaphase FISH
  • mosaicism
  • Prader-Willi syndrome
  • proficiency testing
  • quality assurance
  • SNRPN probe
  • workload

ASJC Scopus subject areas

  • Genetics(clinical)
  • Neuroscience(all)
  • Neuropsychology and Physiological Psychology

Cite this

Dewald, G. W., Stallard, R., Bader, P. I., Chen, K., Zenger-Hain, J., Harris, C. J., ... Yung, J. F. (1996). Toward quality assurance for metaphase FISH: A multicenter experience. American Journal of Medical Genetics, 65(3), 190-196. https://doi.org/10.1002/(SICI)1096-8628(19961028)65:3<190::AID-AJMG4>3.0.CO;2-U

Toward quality assurance for metaphase FISH : A multicenter experience. / Dewald, Gordon W.; Stallard, Richard; Bader, Patricia I.; Chen, Kathy; Zenger-Hain, Julie; Harris, Catherine J.; Higgins, Rodney; Hirsch, Betsy; Hsu, Wei Tong; Johnson, Eric; Kubic, Virginia; Kurczynski, T. W.; Malone, James M.; McCorquodale, D. James; Meilinger, Karen; Meisner, Lorraine F.; Moore, J. W.; Schwartz, Stuart; Siembieda, Steven; Storto, Patrick D.; Vance, Gail; Van Tuinen, Peter; Wiktor, Ann; Yung, Jar Fee.

In: American Journal of Medical Genetics, Vol. 65, No. 3, 28.10.1996, p. 190-196.

Research output: Contribution to journalArticle

Dewald, GW, Stallard, R, Bader, PI, Chen, K, Zenger-Hain, J, Harris, CJ, Higgins, R, Hirsch, B, Hsu, WT, Johnson, E, Kubic, V, Kurczynski, TW, Malone, JM, McCorquodale, DJ, Meilinger, K, Meisner, LF, Moore, JW, Schwartz, S, Siembieda, S, Storto, PD, Vance, G, Van Tuinen, P, Wiktor, A & Yung, JF 1996, 'Toward quality assurance for metaphase FISH: A multicenter experience', American Journal of Medical Genetics, vol. 65, no. 3, pp. 190-196. https://doi.org/10.1002/(SICI)1096-8628(19961028)65:3<190::AID-AJMG4>3.0.CO;2-U
Dewald, Gordon W. ; Stallard, Richard ; Bader, Patricia I. ; Chen, Kathy ; Zenger-Hain, Julie ; Harris, Catherine J. ; Higgins, Rodney ; Hirsch, Betsy ; Hsu, Wei Tong ; Johnson, Eric ; Kubic, Virginia ; Kurczynski, T. W. ; Malone, James M. ; McCorquodale, D. James ; Meilinger, Karen ; Meisner, Lorraine F. ; Moore, J. W. ; Schwartz, Stuart ; Siembieda, Steven ; Storto, Patrick D. ; Vance, Gail ; Van Tuinen, Peter ; Wiktor, Ann ; Yung, Jar Fee. / Toward quality assurance for metaphase FISH : A multicenter experience. In: American Journal of Medical Genetics. 1996 ; Vol. 65, No. 3. pp. 190-196.
@article{eb9040c6d53248e687d262ea57c4adec,
title = "Toward quality assurance for metaphase FISH: A multicenter experience",
abstract = "Although fluorescent in situ hybridization (FISH) is rapidly becoming a part of clinical cytogenetics, no organization sponsors multicenter determinations of the efficacy of probes. We report on 23 laboratories that volunteered to provide slides and to use a probe for small nuclear ribonucleoprotein polypeptide N (SNRPN) and a control locus. Experiences with FISH for these laboratories during 1994 ranged from 0 to 645 utilizations (median = 84) involving blood, amniotic fluid, and bone marrow. In an initial study of hybridization efficiency, the median percentage of metaphases from normal individuals showing two SNRPN and two control signals for slides prepared at each site was 97.0 (range = 74-100); for slides prepared by a central laboratory, it was 97.8 (range = 81.6-100). In a subsequent blind study, each laboratory attempted to score 5 metaphases from each of 23 specimens [8 with del(15)(q11.2→q12) and 15 with normal 15 chromosomes]. Of 529 challenges, the correct SNRPN pattern was found in 5 of 5 metaphases in 457 (86{\%}) and in 4 of 5 in 33 (6{\%}). Ambiguous, incomplete, or no results were reported for 32 (6{\%}) challenges. Seven (1{\%}) diagnostic errors were made, including 6 false positives and 1 false negative: 1 laboratory made 3 errors, 1 made 2, and 2 made 1 each. Most errors and inconsistencies seemed due to inexperience with FISH. The working time to process and analyze slides singly averaged 49.5 min; slides processed in batches of 4 and analyzed singly required 36.9 min. We conclude that proficiency testing for FISH by using an extensive array of challenges is possible and that multiple centers can collaborate to test probes and to evaluate costs.",
keywords = "hybridization efficiency, metaphase FISH, mosaicism, Prader-Willi syndrome, proficiency testing, quality assurance, SNRPN probe, workload",
author = "Dewald, {Gordon W.} and Richard Stallard and Bader, {Patricia I.} and Kathy Chen and Julie Zenger-Hain and Harris, {Catherine J.} and Rodney Higgins and Betsy Hirsch and Hsu, {Wei Tong} and Eric Johnson and Virginia Kubic and Kurczynski, {T. W.} and Malone, {James M.} and McCorquodale, {D. James} and Karen Meilinger and Meisner, {Lorraine F.} and Moore, {J. W.} and Stuart Schwartz and Steven Siembieda and Storto, {Patrick D.} and Gail Vance and {Van Tuinen}, Peter and Ann Wiktor and Yung, {Jar Fee}",
year = "1996",
month = "10",
day = "28",
doi = "10.1002/(SICI)1096-8628(19961028)65:3<190::AID-AJMG4>3.0.CO;2-U",
language = "English (US)",
volume = "65",
pages = "190--196",
journal = "American Journal of Medical Genetics, Part C: Seminars in Medical Genetics",
issn = "1552-4825",
publisher = "Wiley-Liss Inc.",
number = "3",

}

TY - JOUR

T1 - Toward quality assurance for metaphase FISH

T2 - A multicenter experience

AU - Dewald, Gordon W.

AU - Stallard, Richard

AU - Bader, Patricia I.

AU - Chen, Kathy

AU - Zenger-Hain, Julie

AU - Harris, Catherine J.

AU - Higgins, Rodney

AU - Hirsch, Betsy

AU - Hsu, Wei Tong

AU - Johnson, Eric

AU - Kubic, Virginia

AU - Kurczynski, T. W.

AU - Malone, James M.

AU - McCorquodale, D. James

AU - Meilinger, Karen

AU - Meisner, Lorraine F.

AU - Moore, J. W.

AU - Schwartz, Stuart

AU - Siembieda, Steven

AU - Storto, Patrick D.

AU - Vance, Gail

AU - Van Tuinen, Peter

AU - Wiktor, Ann

AU - Yung, Jar Fee

PY - 1996/10/28

Y1 - 1996/10/28

N2 - Although fluorescent in situ hybridization (FISH) is rapidly becoming a part of clinical cytogenetics, no organization sponsors multicenter determinations of the efficacy of probes. We report on 23 laboratories that volunteered to provide slides and to use a probe for small nuclear ribonucleoprotein polypeptide N (SNRPN) and a control locus. Experiences with FISH for these laboratories during 1994 ranged from 0 to 645 utilizations (median = 84) involving blood, amniotic fluid, and bone marrow. In an initial study of hybridization efficiency, the median percentage of metaphases from normal individuals showing two SNRPN and two control signals for slides prepared at each site was 97.0 (range = 74-100); for slides prepared by a central laboratory, it was 97.8 (range = 81.6-100). In a subsequent blind study, each laboratory attempted to score 5 metaphases from each of 23 specimens [8 with del(15)(q11.2→q12) and 15 with normal 15 chromosomes]. Of 529 challenges, the correct SNRPN pattern was found in 5 of 5 metaphases in 457 (86%) and in 4 of 5 in 33 (6%). Ambiguous, incomplete, or no results were reported for 32 (6%) challenges. Seven (1%) diagnostic errors were made, including 6 false positives and 1 false negative: 1 laboratory made 3 errors, 1 made 2, and 2 made 1 each. Most errors and inconsistencies seemed due to inexperience with FISH. The working time to process and analyze slides singly averaged 49.5 min; slides processed in batches of 4 and analyzed singly required 36.9 min. We conclude that proficiency testing for FISH by using an extensive array of challenges is possible and that multiple centers can collaborate to test probes and to evaluate costs.

AB - Although fluorescent in situ hybridization (FISH) is rapidly becoming a part of clinical cytogenetics, no organization sponsors multicenter determinations of the efficacy of probes. We report on 23 laboratories that volunteered to provide slides and to use a probe for small nuclear ribonucleoprotein polypeptide N (SNRPN) and a control locus. Experiences with FISH for these laboratories during 1994 ranged from 0 to 645 utilizations (median = 84) involving blood, amniotic fluid, and bone marrow. In an initial study of hybridization efficiency, the median percentage of metaphases from normal individuals showing two SNRPN and two control signals for slides prepared at each site was 97.0 (range = 74-100); for slides prepared by a central laboratory, it was 97.8 (range = 81.6-100). In a subsequent blind study, each laboratory attempted to score 5 metaphases from each of 23 specimens [8 with del(15)(q11.2→q12) and 15 with normal 15 chromosomes]. Of 529 challenges, the correct SNRPN pattern was found in 5 of 5 metaphases in 457 (86%) and in 4 of 5 in 33 (6%). Ambiguous, incomplete, or no results were reported for 32 (6%) challenges. Seven (1%) diagnostic errors were made, including 6 false positives and 1 false negative: 1 laboratory made 3 errors, 1 made 2, and 2 made 1 each. Most errors and inconsistencies seemed due to inexperience with FISH. The working time to process and analyze slides singly averaged 49.5 min; slides processed in batches of 4 and analyzed singly required 36.9 min. We conclude that proficiency testing for FISH by using an extensive array of challenges is possible and that multiple centers can collaborate to test probes and to evaluate costs.

KW - hybridization efficiency

KW - metaphase FISH

KW - mosaicism

KW - Prader-Willi syndrome

KW - proficiency testing

KW - quality assurance

KW - SNRPN probe

KW - workload

UR - http://www.scopus.com/inward/record.url?scp=0029854178&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029854178&partnerID=8YFLogxK

U2 - 10.1002/(SICI)1096-8628(19961028)65:3<190::AID-AJMG4>3.0.CO;2-U

DO - 10.1002/(SICI)1096-8628(19961028)65:3<190::AID-AJMG4>3.0.CO;2-U

M3 - Article

C2 - 9240742

AN - SCOPUS:0029854178

VL - 65

SP - 190

EP - 196

JO - American Journal of Medical Genetics, Part C: Seminars in Medical Genetics

JF - American Journal of Medical Genetics, Part C: Seminars in Medical Genetics

SN - 1552-4825

IS - 3

ER -