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Development of FISH technology in pathological tissue

Development of FISH technology in pathological tissue
Development of FISH technology in pathological tissue

Fluorescence in situ hybridisation (FISH) and specific DNA probes for peri-centromeric repeat regions and unique sequence loci have made it possible to study chromosomal aberrations from interphase nuclei. Large-scale retrospective studies on the prognostic value of interphase cytogenetics become feasible if these techniques are readily applicable to nuclei from archival formalin-fixed paraffin-embedded tumour tissues. We describe here an improved technique for interphase FISH analysis of tissues that have been fixed in formalin. The suggested protocol has improved probe penetration and hybridisation efficiency by studying individual in situ hybridisation parameters such as tissue section thickness, target unmasking, denaturation, hybridisation, stringency washing, and hybrid detection. This investigation has enabled the formulation of a standard technique which has then been applied in several pathological studies. With minor adaptations the technique has also, been applied to isolated nuclei, from paraffin-embedded tissue sections.

The main aims of this study were to develop the technique of fluorescence in situ hybridisation (FISH) and to study the occurrence of trisomy of chromosome 3 in mucosa-associated lymphoid tissue lymphoma (MALT) and splenic marginal zone cell lymphoma (SMZCL).

The feasibility of using molecular hybridisation techniques for the detection of malignant clones that contain numerical abnormality of chromosome 3 was tested on paraffin-embedded specimens from 58 patients who had MALT lymphoma and 18 patients who had splenic MZCL. A biotinylated DNA probe specific for chromosome 3 was used for in situ hybridisation. We found 19% (11/58) of +3 in patients with MALT lymphoma and in only 5.5% (1/18) of patients with SMZCL. This suggests that trisomy of chromosome 3 is not as common as reported before and can not be considered as a marker for these diseases.

The relationship between low and high grade MALT lymphoma and also between MALT lymphomas and SMZCL is discussed in this study.

University of Southampton
HajMohammadi, Sassan
104a9f4c-e8be-435b-a644-4d33ef980ad3
HajMohammadi, Sassan
104a9f4c-e8be-435b-a644-4d33ef980ad3

HajMohammadi, Sassan (1999) Development of FISH technology in pathological tissue. University of Southampton, Doctoral Thesis.

Record type: Thesis (Doctoral)

Abstract

Fluorescence in situ hybridisation (FISH) and specific DNA probes for peri-centromeric repeat regions and unique sequence loci have made it possible to study chromosomal aberrations from interphase nuclei. Large-scale retrospective studies on the prognostic value of interphase cytogenetics become feasible if these techniques are readily applicable to nuclei from archival formalin-fixed paraffin-embedded tumour tissues. We describe here an improved technique for interphase FISH analysis of tissues that have been fixed in formalin. The suggested protocol has improved probe penetration and hybridisation efficiency by studying individual in situ hybridisation parameters such as tissue section thickness, target unmasking, denaturation, hybridisation, stringency washing, and hybrid detection. This investigation has enabled the formulation of a standard technique which has then been applied in several pathological studies. With minor adaptations the technique has also, been applied to isolated nuclei, from paraffin-embedded tissue sections.

The main aims of this study were to develop the technique of fluorescence in situ hybridisation (FISH) and to study the occurrence of trisomy of chromosome 3 in mucosa-associated lymphoid tissue lymphoma (MALT) and splenic marginal zone cell lymphoma (SMZCL).

The feasibility of using molecular hybridisation techniques for the detection of malignant clones that contain numerical abnormality of chromosome 3 was tested on paraffin-embedded specimens from 58 patients who had MALT lymphoma and 18 patients who had splenic MZCL. A biotinylated DNA probe specific for chromosome 3 was used for in situ hybridisation. We found 19% (11/58) of +3 in patients with MALT lymphoma and in only 5.5% (1/18) of patients with SMZCL. This suggests that trisomy of chromosome 3 is not as common as reported before and can not be considered as a marker for these diseases.

The relationship between low and high grade MALT lymphoma and also between MALT lymphomas and SMZCL is discussed in this study.

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Published date: 1999

Identifiers

Local EPrints ID: 463570
URI: http://eprints.soton.ac.uk/id/eprint/463570
PURE UUID: 048f7424-34fd-4ff7-956b-1b75a9411be7

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Date deposited: 04 Jul 2022 20:53
Last modified: 16 Mar 2024 19:05

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Author: Sassan HajMohammadi

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