Article

FISH Is Superior to PCR in Detecting t(14;18)(q32;q21)-IgH/bcl-2 in Follicular Lymphoma Using Paraffin-Embedded Tissue Samples

Division of Anatomic Pathology, Mayo Clinic, Rochester, MN 55905, USA.
American Journal of Clinical Pathology (Impact Factor: 3.01). 10/2005; 124(3):421-9. DOI: 10.1309/BLH8-MMK8-5UBQ-4K6R
Source: PubMed

ABSTRACT Detection of t(14;18)(q32;q21)-IgH/bcl-2, which is present in 70% to 95% of follicular lymphomas (FLs), might aid in diagnosing FL. The efficacy of routine polymerase chain reaction (PCR) and fluorescence in situ hybridization (FISH) techniques in detecting t(14;18) in paraffin-embedded tissue samples was compared on 5 normal tonsils and 28 FLs demonstrated to be t(14;18)+ by previous karyotyping. There was technical failure in 14 (50%) of the FLs by PCR, likely due to B-5 fixation, and 4 (14%) of FLs by FISH, likely due to advanced specimen age. In the remaining successful cases, 5 (36%) of 14 were positive by PCR and 24 (100%) of 24 were positive by FISH. All 5 normal tonsils were negative by both methods. FISH is superior to PCR for detecting t(14;18) from paraffin-embedded tissue samples because it is more sensitive and equally specific.

Download full-text

Full-text

Available from: Ellen D Remstein, Jul 07, 2015
1 Follower
 · 
108 Views
  • Source
    • "Accordingly , PCR testing for these mutations has become common practice both when first diagnosing many hematologic malignancies and when monitoring for minimal residual disease [Bonassi et al., 1995; Schuler and Dolken, 2006; Jolkowska et al., 2007; Gu et al., 2008]. However, even if the breakpoint locations are well known, as is the case with t(14;18), many reports maintain that FISH is superior to PCR in sensitivity and comparable to PCR in specificity [Einerson et al., 2005; Belaud-Rotureau et al., 2007; Gu et al., 2008]. FISHbased detection of translocations relies on fluorescent DNA probes that bind specifically to selected regions on one or both of the involved chromosomes. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Technical advances have improved the capacity to detect and quantify genetic variants, providing novel methods for the detection of rare mutations and for better understanding the underlying environmental factors and biological mechanisms contributing to mutagenesis. The polymerase chain reaction (PCR) has revolutionized genetic testing and remains central to many of these new techniques for mutation detection. Millions of genetic variations have been discovered across the genome. These variations include germline mutations and polymorphisms, which are inherited in a Mendelian manner and present in all cells, as well as acquired, somatic mutations that differ widely by type and size [from single-base mutations to whole chromosome rearrangements, and including submicroscopic copy number variations (CNVs)]. This review focuses on current methods for assessing acquired somatic mutations in the genome, and it examines their application in molecular epidemiology and sensitive detection and analysis of disease. Although older technologies have been exploited for detecting acquired mutations in cancer and other disease, the high-throughput and high-sensitivity offered by next-generation sequencing (NGS) systems are transforming the discovery of disease-associated acquired mutations by enabling comparative whole-genome sequencing of diseased and healthy tissues from the same individual. Emerging microfluidic technologies are beginning to facilitate single-cell genetic analysis of target variable regions for investigating cell heterogeneity within tumors as well as preclinical detection of disease. The technologies discussed in this review will significantly expand our knowledge of acquired genetic mutations and causative mechanisms.
    Environmental and Molecular Mutagenesis 10/2010; 51(8-9):851-70. DOI:10.1002/em.20630 · 2.55 Impact Factor
  • Source
    • "The bcl-2 protein can be overexpressed or absent immunohistochemically . FISH is preferred usually on paraffin embedded material, applied on interphase nuclei [8], by dual-color fluorescence [9]; certain authors recommend it at diagnosis, as equally sensitive and more specific than PCR [10] [11]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The t(14;18) translocation, which leads to an overproduction of the bcl-2 protein, supposedly occurs in almost all follicular lymphomas (FL) and can be detected by FISH methods or by PCR. Its detection is useful in monitoring the response to therapy and in assessing minimal residual disease in bone marrow. Recently it was observed that the translocation could become negative after treatment. The prognostic and predictive significance of this fluctuation is not entirely understood. We intended to find significant correlations among morphological features, histological grades, immunohistochemical findings, and cytogenetical aberrations in malignant follicular lymphomas, in order to identify the prognostic and predictive value of the bcl-2/IgH translocation in these malignancies. We conducted a study on 79 patients with follicular lymphomas. The study was carried out on tissue samples selected from the "Victor Babes" National Institute of Pathology files. These samples were tested by immunohistochemistry and FISH. Results: Most of the cases (65.2%) were low-grade FL (grade 1-2). Approximately 58.8% of cases in the FISH study group presented t(14;18). In 66.6% of the cases with t(14;18), the immunohistochemical reaction for bcl-2 protein was positive. A significant positive correlation was found between the IHC positivity for bcl-2 and t(14;18) detected by FISH (p=0.04). Bcl-2 t(14;18) plays an important role in the pathogenesis of follicular lymphoma. FISH is an important tool in the diagnosis, treatment and follow up of these malignancies, since the immunohistochemical testing is negative in a significant proportion of cases.
    Romanian journal of morphology and embryology = Revue roumaine de morphologie et embryologie 01/2010; 51(4):687-91. · 0.72 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: We succeed to improve the pulse duration of a diode-pumped KLM Cr:LiSrAlF<sub>6</sub> (Cr:LiSAF) laser using broadband chirped mirrors. Moreover, we apply the laser configuration for a Cr:LiSGaF laser to generate 10-fs pulses. These are, to our knowledge, the shortest pulses ever produced from Cr:LiSAF and Cr:LiSrGaF<sub>6</sub> (Cr:LiSGaF) lasers.
    Lasers and Electro-Optics, 2002. CLEO '02. Technical Digest. Summaries of Papers Presented at the; 02/2002