Translocation detection in lymphoma diagnosis by split-signal FISH: a standardised approach

Rijk, Anke van; Mason, David Y.; Jones, Margaret; Cabeçadas, José; Cigudosa, Juan C.; Garcı́a, Juan F.; Leoncini, Lorenzo; Cocco, Mario; Hansmann, Martin‐Leo; Mottok, Anja; Bergman, Christiane Copie; Baia, Maryse; Anagnostou, Dimitra; Pouliou, Evi; Hamilton-Dutoit, Stephen; Christiansen, Mette Hjøllund; Poulsen, Tim Svenstrup; Matthiesen, Steen H.; Dongen, Jacques J. M. van; Krieken, J. Han van

doi:10.1007/s12308-008-0017-5

Cited by 29 publications

(30 citation statements)

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“…10 Areas of tumor cells used to prepare the TMA were specifically pre-analyzed and confirmed using hematoxylin and eosin-stained slides. Original TMA (from the Euro-FISH program) were centrally prepared using punch needles of 1 mm.…”

Section: Methodsmentioning

confidence: 99%

“…Original TMA (from the Euro-FISH program) were centrally prepared using punch needles of 1 mm. 10 Eighteen months ago, the four different TMA were stained with 16 different FISH probes (see Table 1) in eight European laboratories (4 probes per laboratory resulting in 2 independent duplicates per probe), analyzed and subsequently stored in the dark at 4°C and normal air pressure. Fifteen TMA, each stained with a different CISH-compatible FISH probe (with the lowest number of lost cores per probe after FISH staining), were chosen for CISH staining.…”

Section: Methodsmentioning

confidence: 99%

“…In addition to these slides, one slide cut from TMA n. 3 (kindly provided by the Euro-FISH consortium), stored unstained for the same period of time and under the same conditions as described above, was freshly stained for BCL6-FISH and CISH. Furthermore, a metaphase spread of B-lymphocytes from a healthy donor, used during the Euro-FISH program to validate the BCL6 FISH-probe, 10 was re-used for DuoCISH staining. CISHstained slides were analyzed by bright-field microscopy.…”

Section: Methodsmentioning

confidence: 99%

“…All previously split-signal FISH-stained slides had been stored in a dark room at 4°C (normal air pressure). One slide, stored unstained at 4°C, was freshly BCL6-stained using the FISH protocol previously described 10 and subsequently DuoCISH-stained. The following FISH probes (with chromosomal localization), serving as templates for DuoCISH-staining, were used: BCL10(1p22); IGK(2p11); ALK(2p23); BCL6(3q27); TCRG(7p14); TCRB(7q34); MYC(8q24); PAX5(9p13); CCND1(11q13); TCRAD(14q11); TCL1(14q32); IGH(14q32); MALT1(18q21); BCL2(18q21); BCL3(19q13); and IGL (22q11).…”

Section: Probes and Fluorescence And Chromogenic In Situ Hybridizatiomentioning

confidence: 99%

“…10 In this study we describe double-staining chromogenic in situ hybridization (DuoCISH) as an alternative to split-signal FISH in the diagnosis of lymphoma.…”

Section: Introductionmentioning

confidence: 99%

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Double-staining chromogenic in situ hybridization as a useful alternative to split-signal fluorescence in situ hybridization in lymphoma diagnostics

Rijk¹,

Poulsen²,

Jones³

et al. 2009

Haematologica

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BackgroundMalignant lymphomas are classified based on morphology, immunophenotype, genetics and clinical features. The pathological diagnosis is generally considered difficult and prone to mistakes. Since non-random chromosomal translocations are specifically involved in specific entities, their detection is an important adjunct for increasing the reliability of the diagnosis. Recently, split-signal fluorescence in situ hybridization has become available as a robust method to detect chromosomal breaks in paraffin-embedded formalin-fixed tissues. A bright field approach would bring this technology within the reach of every pathology laboratory. Design and MethodsOur study was initiated to determine the consistency between chromogenic in situ hybridization and fluorescence in situ hybridization, both using split-signal probes developed for the detection of chromosomal breaks. Five hundred and forty cases of 11 lymphoma entities and reactive, benign lymphoid tissues, collected from eight different pathology laboratories, placed on 15 fluorescence in situ hybridization pre-stained tissue microarray slides, were double stained for the chromogenic hybridization. For each core morphology and actual signal were compared to the original fluorescence hybridization results. In addition, hematoxylin background staining intensity and signal intensity of the double-staining chromogenic in situ hybridization procedure were analyzed. ResultsWith respect to the presence or absence of chromosomal breaks, 97% concordance was found between the results of the two techniques. Hematoxylin background staining intensity and signal intensity were found to correspond. The overall morphology after doublestaining chromogenic in situ hybridization had decreased compared to the initial morphology scored after split-signal fluorescence in situ hybridization staining. ConclusionsWe conclude that double-staining chromogenic in situ hybridization is equally reliable as fluorescence in situ hybridization in detecting chromosomal breaks in lymphoid tissue. Although differences in morphology, hematoxylin staining and chromogenic signal intensity vary between the tumor entities none of the entities appeared more easy or difficult to score.Key words: double staining, CISH, split-signal, lymphoma diagnostics.Citation: van Rijk A, Svenstroup-Poulsen T, Jones M, Cabeçadas J, Cigudosa JC, Leoncini L, Mottok A, Bergman CC, Pouliou E, Hamilton Dutoit S, and van Krieken HJ. Double-staining chromogenic in situ hybridization as a useful alternative to split-signal fluorescence in situ hybridization in lymphoma diagnostics. Haematologica. 2010;95:247-252.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Probes and Fluorescence And Chromogenic In Situ Hybridizatiomentioning

confidence: 99%

“…10 In this study we describe double-staining chromogenic in situ hybridization (DuoCISH) as an alternative to split-signal FISH in the diagnosis of lymphoma.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Double-staining chromogenic in situ hybridization as a useful alternative to split-signal fluorescence in situ hybridization in lymphoma diagnostics

Rijk¹,

Poulsen²,

Jones³

et al. 2009

Haematologica

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Confirmation of a Mutation by Multiple Molecular Approaches

Martinez‐Valdez

Ortiz‐Quintero

2011

Gene Discovery for Disease Models

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Immunoglobulin heavy‐chain fluorescence in situ hybridization‐chromogenic in situ hybridization DNA probe split signal in the clonality assessment of lymphoproliferative processes on cytological samples

Zeppa

Fernandez²,

Cozzolino³

et al. 2012

Cancer Cytopathology

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BACKGROUND:The human immunoglobulin heavy-chain (IGH) locus at chromosome 14q32 is frequently involved in different translocations of non-Hodgkin lymphoma (NHL), and the detection of any breakage involving the IGH locus should identify a B-cell NHL. The split-signal IGH fluorescence in situ hybridization-chromogenic in situ hybridization (FISH-CISH) DNA probe is a mixture of 2 fluorochrome-labeled DNAs: a green one that binds the telomeric segment and a red one that binds the centromeric segment, both on the IGH breakpoint. In the current study, the authors tested the capability of the IGH FISH-CISH DNA probe to detect IGH translocations and diagnose B-cell lymphoproliferative processes on cytological samples. METHODS: Fifty cytological specimens from cases of lymphoproliferative processes were tested using the split-signal IGH FISH-CISH DNA probe and the results were compared with light-chain assessment by flow cytometry (FC), IGH status was tested by polymerase chain reaction (PCR), and clinicohistological data. RESULTS: The signal score produced comparable results on FISH and CISH analysis and detected 29 positive, 15 negative, and 6 inadequate cases; there were 29 true-positive cases (66%), 9 true-negative cases (20%), 6 false-negative cases (14%), and no false-positive cases (0%). Comparing the sensitivity of the IGH FISH-CISH DNA split probe with FC and PCR, the highest sensitivity was obtained by FC, followed by FISH-CISH and PCR. CONCLUSIONS: The split-signal IGH FISH-CISH DNA probe is effective in detecting any translocation involving the IGH locus. This probe can be used on different samples from different B-cell lymphoproliferative processes, although it is not useful for classifying specific entities. Cancer (Cancer Cytopathol) 2012;120:390-400. V C 2012 American Cancer Society.KEY WORDS: immunoglobulin heavy-chain (IGH), fluorescence in situ hybridization-chromogenic in situ hybridization (FISH-CISH), DNA split probe, flow cytometry, polymerase chain reaction, lymphoproliferative processes, cytology.

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Translocation detection in lymphoma diagnosis by split-signal FISH: a standardised approach

Cited by 29 publications

References 11 publications

Double-staining chromogenic in situ hybridization as a useful alternative to split-signal fluorescence in situ hybridization in lymphoma diagnostics

Double-staining chromogenic in situ hybridization as a useful alternative to split-signal fluorescence in situ hybridization in lymphoma diagnostics

Confirmation of a Mutation by Multiple Molecular Approaches

Immunoglobulin heavy‐chain fluorescence in situ hybridization‐chromogenic in situ hybridization DNA probe split signal in the clonality assessment of lymphoproliferative processes on cytological samples

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