Analysis of BCR/ABL Abnormalities in mRNA from 20-Year-Old Paraffin-Embedded Tissue for BCR/ABL Rearrangement by Polymerase Chain Reaction

Aurer, Igor; Juhbashi, Tohru; Sekine, Ichiro; Tomonaga, Masao; Gale, Robert Peter

doi:10.1159/000204364

Cited by 12 publications

(9 citation statements)

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“…A model disease where molecular studies are of significant practical relevance is chronic myelogenous leukemia (CML), which is characterized by the hallmark Philadelphia chromosome caused by the t(9;22) translocation resulting in a breakpoint cluster region-Abelson (BCR-ABL) fusion. Already more than 10 years ago, up to two-decade-old archival tissues were demonstrated to be suitable for detection of BCR-ABL hybrid genes and fusion transcripts [3]. In accordance with these data, the successful detection of BCR-ABL transcripts also has been shown in other reports [40,41].…”

Section: Molecular Diagnostics In Myeloproliferative Disorderssupporting

confidence: 68%

Ancillary techniques in bone marrow pathology: molecular diagnostics on bone marrow trephine biopsies

et al. 2005

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Pathologic examination of trephine bone marrow (BM) biopsies plays a central role in the diagnosis and staging of haematological neoplasms and other disorders affecting haematopoiesis. Haematopathology has been profoundly influenced by the advent of molecular genetic techniques suitable for paraffin-embedded tissues, and certain applications, such as the determination of B- and T-cell clonality, belong to its standard diagnostic repertoire. Many of these molecular tests can be performed successfully with nucleic acids extracted from BM trephine biopsies, if some technical aspects specific to this template source such as various fixation and decalcification procedures are taken into consideration. The current indications for molecular BM diagnostics range from the confirmation of lymphoma involvement with gene rearrangement analysis, demonstration of tumor-specific translocations in lymphoid and chronic myeloproliferative disorders along to the detection of microorganisms or marrow involvement by soft tissue sarcomas. The availability of quantitative polymerase chain reaction techniques for the investigation of allelic imbalances and gene expression levels in paraffin-embedded material also open new avenues for research and advanced diagnostics. The molecular detection of minimal residual disease in haematological neoplasms, especially in the context of new treatment strategies, will provide future challenges. This article summarizes the current state of the art in molecular diagnostics applied to paraffin-embedded BM biopsies.

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Section: Molecular Diagnostics In Myeloproliferative Disorderssupporting

confidence: 68%

Ancillary techniques in bone marrow pathology: molecular diagnostics on bone marrow trephine biopsies

et al. 2005

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“…However, RT-PCR amplification of RNA from this material is a complex procedure that may be adversely influenced by several factors, mainly by nucleic acid breakdown during tissue processing and storage (Aurer et al, 1993). A recent extensive analysis of human and viral genes in FFPE tissue sections showed that amplification can be achieved most consistently by using primers that encompass small (less than 200 bp) RNA targets .…”

Section: Soguero Et Almentioning

confidence: 99%

Assessment of Genotype and Molecular Evolution of Hepatitis C Virus in Formalin-Fixed Paraffin-Embedded Liver Tissue from Patients With Chronic Hepatitis C Virus Infection

Soguero

Campo

Ribalta

et al. 2000

Laboratory Investigation

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SUMMARY:Drawbacks of hepatitis C virus (HCV) RNA detection in paraffin-embedded liver tissue have satisfactorily been solved by RT-PCR amplification of the 5Јnon-coding region (5ЈNCR). However, detection of this highly conserved region does not provide information on epidemiological or pathogenetic aspects of HCV infection. This study explores whether other functionally important genetic regions of HCV, such as the hypervariable region 1 (HVR-1) and the interferon sensitivity-determining region (ISDR), can be retrieved from paraffin-embedded liver specimens by RT-PCR, and whether the amplified material is suitable for further molecular analyses. RT-PCR amplification of 5ЈNCR, HVR-1, and ISDR was assessed in RNA extracted from 50 formalin-fixed, paraffin-embedded liver specimens, including 23 needle liver biopsies (11 from patients with non-A, non-B chronic hepatitis diagnosed between 1971 and 1985, 8 from subjects with normal liver histology and 4 from sequential biopsies from a patient with HCV recurrence after liver transplantation), and 27 liver explants from patients undergoing transplantation between 1988 and 1996 (16 with HCV-related cirrhosis and 11 with other disorders). The 5ЈNCR was successfully amplified in 8 of 11 (73%) non-A, non-B chronic hepatitis biopsies and in all of the specimens from patients with serological documentation of HCV infection. There were no false-positive results. HCV genotype was identified by RFLP analysis of the 5ЈNCR in the 13 cases analyzed. HVR-1 and ISDR were amplified in 24 of 28 (86%) samples, which were positive for the 5ЈNCR. Efficient amplification was inversely related to the time of storage. The evolutionary changes of HVR-1 and ISDR were successfully analyzed by direct sequencing of amplificates from the explanted liver and from the sequential liver biopsies in a patient with HCV infection recurrence after transplantation. These observations indicate that paraffin-embedded liver tissue, even when stored for more than 20 years, is appropriate for advanced studies on the molecular biology of HCV. (Lab Invest 2000, 80:851-856).

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“…The ability of ionizing radiations to cause contiguous double-stand DNA breaks proximity of BCR and ABL1 in the interphase nucleus and detection of BCRABL1 transcripts in leukemia cells from atomic bomb survivors with CML. CML developing in these persons began when they were exposed to radiations from the A-bombs [3,4]. If we accept this hypothesis we can estimate (minimally) the interval from developing BCR-ABL1 to the diagnosis of chronic phase CML.…”

Section: Introductionmentioning

confidence: 99%

Therapy-free remission in chronic myeloid leukemia: possible mechanism

Gale

Hochhaus

2018

Expert Review of Hematology

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Analysis of BCR/ABL Abnormalities in mRNA from 20-Year-Old Paraffin-Embedded Tissue for BCR/ABL Rearrangement by Polymerase Chain Reaction

Cited by 12 publications

References 3 publications

Ancillary techniques in bone marrow pathology: molecular diagnostics on bone marrow trephine biopsies

Ancillary techniques in bone marrow pathology: molecular diagnostics on bone marrow trephine biopsies

Assessment of Genotype and Molecular Evolution of Hepatitis C Virus in Formalin-Fixed Paraffin-Embedded Liver Tissue from Patients With Chronic Hepatitis C Virus Infection

Therapy-free remission in chronic myeloid leukemia: possible mechanism

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