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  • 1
    ISSN: 1569-8041
    Keywords: anaplastic large-cell lymphoma ; fluorescent in situ hybridization ; Hodgkin's disease ; Ki-1 lymphoma ; PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: Anaplastic large-cell lymphoma (ALCL) is a recently recognized, distinctive type of non-Hodgkin's lymphoma characterized by anaplastic large-cell cytology and expression of a member of the TNF-receptor family CD30. A characteristic chromosomal translocation has been identified in ALCL of T- or null-cell lineage which juxtaposes a novel tyrosine kinase (anaplastic lymphoma kinase, ALK) located at 2p23 with the nucleophosmin gene (NPM) at 5q35. A chimeric mRNA transcript is produced, and the translocation results in constitutive expression of a truncated form of the ALK protein, p80. There is controversy concerning whether or not the translocation occurs in Hodgkin's disease. The aim of this study was to develop a methodology for fluorescent in situ hybridization (FISH) to detect the t(2;5)(p23;q35), and to compare the results with conventional cytogenetics, reverse-transcriptase PCR and immunostaining for the p80protein. Patients and methods: Twenty-five cases of malignant lymphoma (11 ALCL and 14 HD) were studied. Immunohistochemistry was performed to confirm the diagnosis and for analysis of p80 expression. Conventional cytogenetics were analyzed on G-banded metaphase spreads. FISH was performed using whole chromosome paints for chromosomes 2 and 5 on metaphase spreads and YAC probes for inter phase nuclei. Reverse-transcriptase PCR using primers for ALK and NPM was used to amplify the translocation breakpoint in extracted mRNA. Results: Among 11 cases of ALCL examined by FISH, the translocation was detected in 4. Two of these cases also had RT-PCR and p80 staining performed, with positive results. Among 7 cases where the t(2;5) was not detected by FISH, 3 cases were examined by RT-PCR with negative results and4 cases by p80 staining, also negative. The RT-PCR was negative in all 14cases of Hodgkin's disease, 4 of which were also examined by FISH and found to be negative. Conclusion: Fluorescent in situ hybridization is useful method for detection of the t(2;5)(p23;q35) in anaplastic large-cell lymphoma. The results concur with those of RT-PCR for the chimeric transcript and immunostaining for the p80 protein. The frequency with which the translocation was found was 36% in this small series, and no evidence of the translocation was found in cases of Hodgkin's disease.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1437-1596
    Keywords: DNA ; PCR ; Short tandem repeats ; Forensic Identification ; DNA ; PCR ; Short tandem repeats ; Forensische Identifikation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine , Law
    Description / Table of Contents: Zusammenfassung Eine Quadruplex-Reaktion wurde entwickelt, die die Short tandem repeat loci (STR) HUMVWA31/1, HUMTHO1, HUMF13A1 und HUMFES/FPS amplifiziert. Zum Nachweis der PCR-Produkte werden denaturierende Polyacrylamid-Gele benutzt, in Kombination mit einer Fluoreszenz-basierten Technologie. Dieses System wurde fÜr den Gebrauch in der routinemäßigen Fallbearbeitung evaluiert und hat sich als robust und reproduzierbar erwiesen. Die Quadruplex-Reaktion ist genau so empfindlich wie das kommerzielle HLADQα-Ampli-type Typisierungssystem und kann sowohl an degradiertem als auch an gealtertem Material verwendet werden. Die Probleme einer umgebungsbedingten Kontamination haben sich als begrenzt erwiesen, vorausgesetzt, daß strikte prozedurale Praktiken benutzt werden, d. h. physische Trennung der Extraktion und der amplifizierten Produkte-, Benutzung persönlicher Einrichtungen, wie Pipetten; Abtrennung des Bereichs der Amplifikationsvorbereitung; Benutzung der Laminar Flow-Einrichtung für die Vorbereitung der Amplifikation. Die FÄhigkeit des Systems, Mischungen zu erkennen und die erfolgreiche Analyse von Spurenfällen haben gezeigt, daß dieses System gut geeignet ist als ein Werkzeug für forensische Untersuchung.
    Notes: Abstract A quadruplex reaction has been developed which amplifies the short tandem repeat (STR) loci HUMVWA31/A, HUMTHO1, HUMF13A1 and HUMFES/FPS. Detection of the PCR products employs denaturing poly-acrylamide gels coupled with fluorescent-based technology. This system has been evaluated for use in routine forensic casework and has been shown to be both robust and reproducible. The quadruplex reaction is as sensitive as the commercially available HLA DQα Amplitype typing system and can be used on both degraded and aged material. The problems of environmental contamination have been shown to be limited provided strict procedural practices are followed — i.e. physical separation of sample extraction and amplified products; the use of dedicated equipment such as pipettes; the separation of amplification preparation area. The ability of the system to detect mixtures and the successful analysis of case stains has shown that this system is well suited as a tool for forensic investigation.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1569-8041
    Keywords: anaplastic large-cell lymphoma ; fluorescent in situ hybridization ; Hodgkin's disease ; Ki-1 lymphoma ; PCR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Background: Anaplastic large-cell lymphoma (ALCL) is a recentlyrecognized, distinctive type of non-Hodgkin's lymphoma characterized byanaplastic large-cell cytology and expression of a member of theTNF-receptor family CD30. A characteristic chromosomal translocation hasbeen identified in ALCL of T- or null-cell lineage which juxtaposes a noveltyrosine kinase (anaplastic lymphoma kinase, ALK) located at 2p23 with thenucleophosmin gene (NPM) at 5q35. A chimeric mRNA transcript is produced,and the translocation results in constitutive expression of a truncated formof the ALK protein, p80. There is controversy concerning whether or not thetranslocation occurs in Hodgkin's disease. The aim of this study was todevelop a methodology for fluorescent in situ hybridization (FISH) to detectthe t(2;5)(p23;q35), and to compare the results with conventionalcytogenetics, reverse-transcriptase PCR and immunostaining for the p80protein. Patients and methods: Twenty-five cases of malignant lymphoma (11 ALCLand 14 HD) were studied. Immunohistochemistry was performed to confirm thediagnosis and for analysis of p80 expression. Conventional cytogenetics wereanalyzed on G-banded metaphase spreads. FISH was performed using wholechromosome paints for chromosomes 2 and 5 on metaphase spreads and YACprobes for interphase nuclei. Reverse-transcriptase PCR using primers forALK and NPM was used to amplify the translocation breakpoint in extractedmRNA. Results: Among 11 cases of ALCL examined by FISH, the translocation wasdetected in 4. Two of these cases also had RT-PCR and p80 stainingperformed, with positive results. Among 7 cases where the t(2;5) was notdetected by FISH, 3 cases were examined by RT-PCR with negative results and4 cases by p80 staining, also negative. The RT-PCR was negative in all 14cases of Hodgkin's disease, 4 of which were also examined by FISH and foundto be negative. Conclusion: Fluorescent in situ hybridization is useful methodfor detection of the t(2;5)(p23;q35) in anaplastic large-cell lymphoma. Theresults concur with those of RT-PCR for the chimeric transcript andimmunostaining for the p80 protein. The frequency with which the translocationwas found was 36% in this small series, and no evidence of thetranslocation was found in cases of Hodgkin's disease.
    Type of Medium: Electronic Resource
    Library Location Call Number Volume/Issue/Year Availability
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