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The metaphase FISH image (patient No. 13) shows two chromosomes 5 of apparently identical size, marked by the green centromeric signals. The lack of the 5q31 signal in one of the two chromosomes demonstrates the presence of a small deletion not detectable by conventional karyotyping. Hybridization efficiency was documented by the normal hybridization pattern on the two chromosomes 7 displaying red signals on the centromere and at the 7q31 band. The two red signals on the chromosome 7 in the middle of the figure are close to each other possibly due to crossing with another chromosome.
Source publication
At diagnosis, approximately half of myelodysplastic (MDS) patients presents a normal karyotype by conventional cytogenetic analysis (CCA). Fluorescent in situ hybridization (FISH) is more sensitive than CCA allowing for the detection of minor clones and of submicroscopic lesions. We have analyzed by FISH 101 MDS patients with normal karyotype for t...
Contexts in source publication
Context 1
... results are shown in Table 4. At diagnosis, trisomy 8 was found in few mitotic figures of too poor quality for karyotyping (fuzzy and overlapping chromosomes) in four out of six patients (Nos 1, 2, 3, 4) with Leukemia apparently normal karyotype and trisomy 8 by FISH interphase analysis. ...
Context 2
... mitotic cell with −7 could be visualized by metaphase FISH in patient No. 7. Metaphase FISH analysis showed the presence of a clone carrying a submicroscopic deletion of chromosome 5q31 (Figure 1) in four out of five cases (Nos 9, 10, 12, 13, 14) with an apparently normal chromosome 5 pair. Likewise, a minor clone with submicroscopic 7q31 deletion in metaphase cells was found by metaphase FISH in three out of five patients with 7q−. ...
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Citations
... FISH may complement conventional cytogenetic analysis in situations of failure of standard G-banding (absent or poor-quality metaphases). FISH analysis of del(5q) or with del(7q) or monosomy 7 may provide prognostic information [71,72]. Targeted FISH for deletion 5q may be indicated for testing, as it is a recognized WHO classification entity of MDS with low blasts and isolated 5q deletion. ...
... Although FISH is specific, with limited sensitivity, it is important to recognize that FISH can only be applied in a targeted fashion; hence, comprehensive assessment for chromosomal aberrations cannot be carried out using this technique due to the spectrum of recurrent chromosome abnormalities for MDS. However, FISH may be useful for clarifying complex aberrations, and it can detect abnormalities in up to 15% of karyotypically normal MDS patients [71][72][73]. ...
Simple Summary
Myelodysplastic Neoplasms (MDS) are a type of blood cancer presenting as ineffective production of blood cells, even though the bone marrow appears active. Traditionally, the response to treatment has been studied by examining blood cell counts, morphologic features, and chromosomal changes. Recently, the ability to evaluate neoplastic status has been significantly improved by the development of highly sensitive flow cytometry and molecular assays, allowing for the identification of residual low-level neoplastic components. In this review, we discuss the evolving concept of measurable (minimal) residual disease (MRD) in MDS in clinical practice, elaborate on the laboratory methods utilized to identify low-level neoplasms, and provide an overview of the published studies correlating MRD with the clinical outcomes of MDS patients.
Abstract
Myelodysplastic Neoplasms (MDS) have been traditionally studied through the assessment of blood counts, cytogenetics, and morphology. In recent years, the introduction of molecular assays has improved our ability to diagnose MDS. The role of Measurable (minimal) Residual Disease (MRD) in MDS is evolving, and molecular and flow cytometry techniques have been used in several studies. In this review, we will highlight the evolving concept of MRD in MDS, outline the various techniques utilized, and provide an overview of the studies reporting MRD and the correlation with outcomes.
... No que diz respeito à citogenética molecular, em geral é possível a realização de uma análise mais sensível, capaz de identificar e investigar pequenas deleções e alterações intersticiais nos cromossomos que não são detectáveis pelos métodos convencionais (SECKER-WALKER et al., 1998;RIGOLIN et al., 2001;BERNASCONI et al., 2003). ...
... Several studies evaluating the added value of FISH to CC have generated mixed results. [7][8][9][10][11][12][13][14][15][16][17] In this study, we evaluated 127 patients with a presumptive diagnosis of MDS using CC and FISH to assess the additional diagnostic and prognostic yield over conventional karyotyping alone. ...
... Several studies have suggested a significant role for FISH analysis, especially in chromosomally normal patients with MDS/AML. [9][10][11] In a study of 57 chromosomally normal patients with MDS, Bernasconi et al. [9] detected occult chromosomal abnormalities by FISH in 15% of patients, resulting in a change in IPSS for five of the nine patients. Furthermore, FISH positivity was associated with an eightfold increase in progression to advanced MDS or AML. ...
... Furthermore, FISH positivity was associated with an eightfold increase in progression to advanced MDS or AML. These findings were corroborated by Rigolin et al. [10] whose analysis of 101 consecutive patients of MDS with normal karyotypes found occult chromosomal abnormalities by FISH in 18 patients (17.8%) and were associated with higher risk disease and worse outcomes as compared to FISH-negative patients. Of note, a third of the FISH-positive patients showed trisomy 8-a chromosomal abnormality-that is AML-MRC = acute myeloid leukemia with myelodysplastic-related changes, CMML = chronic myelomonocytic leukemia, FISH = fluorescence in situ hybridization, MDS = myelodysplastic syndrome, MDS-EB1 = myelodysplastic syndrome with excess blasts type 1, MDS-EB2 = myelodysplastic syndrome with excess blasts type 2, MDS-MLD = myelodysplastic syndrome with multilineage dysplasia, MDS w/isolated del(5q) = myelodysplastic syndrome with isolated deletion 5q, Neg = negative, RCC = refractory cytopenia of childhood, TR-MDS = therapy-related myelodysplastic syndrome no longer considered disease defining, though remains part of the IPSS-R cytogenetic risk stratification schema. ...
Background
Myelodysplastic syndromes (MDSs) are a heterogeneous group of clonal hematopoietic neoplasms, roughly half of which harbor cytogenetic abnormalities with diagnostic, prognostic, and therapeutic significance. Fluorescence in situ hybridization (FISH) for the most commonly seen abnormalities (5/5q, –7/7q, +8, and –20/20q–) is routinely performed alongside conventional cytogenetics (CC) in the evaluation of suspected MDS despite conflicting reports of its relative contribution compared to CC alone.
Objectives
To assess the additional diagnostic and prognostic value of performing concurrent FISH versus CC alone in cases of suspected MDS.
Materials and Methods
A total of 127 bone marrow samples submitted to our cytogenetic laboratory with a presumptive diagnosis of MDS were evaluated by concurrent CC and an MDS FISH panel.
Results
CC was used as the gold standard method with 100% sensitivity in detecting suspected MDS-associated cytogenetic abnormalities. FISH alone had a sensitivity of 76%, whereas CC alone achieved a sensitivity of 97%. The addition of FISH did not change the diagnosis nor change the Revised International Prognostic Scoring System score in any patient. Moreover, in 12 cases identified as positive by both CC and FISH, CC identified multiple chromosomal aberrations of clinical significance not interrogated by the FISH probe panel.
Conclusion
CC alone is sufficiently sensitive in detecting suspected MDS-associated cytogenetic abnormalities that influence clinical decision-making. Routine FISH testing does not provide a significant increase in test sensitivity when an adequate karyotype is obtained. Therefore, FISH testing is best reserved for suspected MDS cases lacking sufficient metaphases.
... 3,5,9,10 The cytogenetic analysis of peripheral blood for diagnosis and disease management provides a less-invasive method instead of bone marrow aspiration, but the utility of peripheral blood to use in FISH and specially karyotype is not clear and additional studies can provide further information to compare the cytogenetic of BM and PB. 1 Several studies compared the results of FISH and karyotype methods using BM samples, but in this study, we compared the results of karyotype and FISH in each of the BM and PB specimens. 8,[11][12][13][14][15][16][17] As karyotype is the most popular method for cytogenetic abnormality assessment, the survey in this field can be useful. 5,18 In this study, we applied optimized karyotype for PB samples. ...
... These studies determined the most useful probes in MDS; however, the majority of these studies reported the superiority of FISH method in about 10%-20% of cases. 4,7,[11][12][13][14][15][16][17][18] In this study, the most common MDS-associated FISH panel probe was used and the results showed As already mentioned, the concordance between BM and PB sample using FISH method was appropriate, but the rate of abnormality detection using FISH method is not appropriate in both types of samples. As a result, examining peripheral blood using FISH method seems to be useful in the patient with known abnormality, and this usually occurs in disease monitoring and it could be useful to avoid serial bone marrow aspiration. ...
Objective
To clear the role of peripheral blood as a substitution for bone marrow in myelodysplastic syndrome and to evaluate the concordance between peripheral blood and bone marrow using karyotype and fluorescence in situ hybridization (FISH) methods.
Methods
We examined 35 bone marrow (BM) and peripheral blood (PB) samples from myelodysplastic syndrome (MDS) patient using karyotype and FISH. Karyotype method for BM and PB samples performed using the standard protocol with an exception for peripheral blood in which growth factor for cultivation was not used. FISH testing was performed using a panel of MDS‐associated probes to detect 20q12, 20qter, 5q31, 5q33, 5p15 and chromosome 7 and 8 centromeres.
Results
Our results showed karyotypes of BM and PB are concordant in 74% of cases, while about 53% of these concordances were achieved from cases with normal karyotypes. However, the results of BM FISH were completely concordant with PB FISH.
Conclusion
Although peripheral blood karyotype is not trustworthy for MDS diagnosis, examining peripheral blood, using the FISH method, could be useful for clinical monitoring.
... A lower number of MP analyzed (< 20) is associated with a higher chance of missing small clones [2]. The prognostic relevance of these smaller clones remains to be elucidated in lower-risk MDS [4]. ...
... Unfortunately, the impact of smaller clone sizes was not assessed in that study, similar to a large important study, performed by Schanz, et al. [10]. A study of 101 MDS patients with normal karyotype revealed small clones (ranging 15% to 32%) using FISH techniques in 18 patients [4]. FISH abnormalities were predictive for worse prognosis, but the majority of these 18 patients had higher-risk MDS and all three patients with refractory anemia were surviving at time of evaluation. ...
Conventional karyotype is one of the most relevant prognostic factors in MDS. However, about 50% of patients with MDS have a normal karyotype. Usually, 20-25 normal metaphases (nMP) are considered to be optimal to exclude small abnormal clones which might be associated with poor prognosis. This study evaluated the impact of examining a suboptimal number of metaphases in patients recruited to the EUMDS Registry with low and intermediate-1 risk according to IPSS. Only 179/1049 (17%) of patients with a normal karyotype had a suboptimal number of nMP, defined as less than 20 metaphases analyzed. The outcome (overall survival and progression-free survival) of patients with suboptimal nMP was not inferior to those with higher numbers of analyzed MP both in univariate and multivariate analyses. For patients with an abnormal karyotype, 224/649 (35%) had a suboptimal number of MP assessed, but this did not impact on outcome. For patients with a normal karyotype and suboptimal numbers of analyzable metaphases standard evaluation might be acceptable for general practice, but we recommend additional FISH-analyses or molecular techniques, especially in candidates for intensive interventions.
... Furthermore, a recent report validated the use of FISH on peripheral blood CD34+ cells in assigning cytogenetic risk classification per IPSS and IPSS-R risk groups, suggesting a role for FISH when chromosomal banding analysis of bone marrow samples is not possible [54]. Some studies illustrate discovery of abnormalities by FISH having association with inferior prognosis in a significant proportion of patients with normal metaphase cytogenetics (MC) [55,58] compared to other studies demonstrating much lower yield [56,57].F o r now, in cases of adequate metaphase cells, FISH likely has little to add and perhaps should not be routinely ordered on every bone marrow sample [59]. Although cytogenetics is the most important prognostic factor in MDS, approximately 50% of de novo MDS patients do not have detectable aberrations or have inadequate karyotyping using MC [44].About three-quarters of patients fall into the 'good' prognostic subgroups of the IPSS and IPSS-R, respectively, largely due to the high prevalence of normal cytogenetics [26]. ...
... Furthermore, a recent report validated the use of FISH on peripheral blood CD34+ cells in assigning cytogenetic risk classification per IPSS and IPSS-R risk groups, suggesting a role for FISH when chromosomal banding analysis of bone marrow samples is not possible [54]. Some studies illustrate discovery of abnormalities by FISH having association with inferior prognosis in a significant proportion of patients with normal metaphase cytogenetics (MC) [55,58] compared to other studies demonstrating much lower yield [56,57]. For now, in cases of adequate metaphase cells, FISH likely has little to add and perhaps should not be routinely ordered on every bone marrow sample [59]. ...
Evaluation of: Bejar R, Lord A, Stevenson K, et al. TET2 mutations predict response to hypomethylating agents in myelodysplastic syndrome patients. Blood 2014 Oct 23;124(17):2705-12. Patients with myelodysplastic syndromes (MDS) have clinically variable courses even within the same prognostic subgroups. Although hypomethylating agents (HMAs) have been shown to improve outcomes in patients with high-risk MDS, many patients do not derive benefit. There is an urgent clinical need to identify patients with low probability of benefiting from HMAs but no reliable clinical predictors or biomarkers have been discovered to date. Although some recurrent molecular mutations in MDS carry independent prognostic value, their ability to predict benefit from HMAs is not clear. Here, we discuss an important article in which sequencing from samples of 213 patients identified recurrent mutations associated with response to HMAs. Although an important step in the right direction, the clinical implications of these findings are far from optimal and identification of biomarkers that can reliably predict benefit from HMAs and other therapies in patients with MDS remains a top clinical and a research priority.
... Most chromosomal aberrations in MDS can also be detected by fluorescence in situ hybridization (FISH) analyses, but only pre-defined anomalies can be covered, if a distinct informative probe is used. 9 The IPSS/-R is based on chromosome banding analyses in primary untreated MDS patients. 3,6,7 If a bone marrow aspiration is impossible or unsuccessful, e.g. because of dry marrow without liquid BM blood, a lack of informative karyotyping because of metaphases failure or the patient´s refusal (5%-20%), 10-12 a reliable karyotyping and thus an adequate cytogenetic risk classification, and, finally, assessment of IPSS/-R risk groups are not possible. ...
International Prognostic Scoring Systems are used to determine the individual risk profile of myelodysplastic syndrome patients. For the assessment of International Prognostic Scoring Systems, an adequate chromosome banding analysis of the bone marrow is essential. Cytogenetic information is not available for a substantial number of patients (5-20%) with dry marrow or an insufficient number of metaphase cells. For these patients, a valid risk classification is impossible. In the study presented here, the International Prognostic Scoring Systems were validated based on Fluorescence-in-situ-hybridization analyses using extended probe panels applied to cluster of differentiation 34 positive (CD34+) peripheral blood cells of 328 MDS patients of our prospective multicenter German diagnostic study and compared to chromosome banding results of 2902 previously published patients with myelodysplastic syndromes: For cytogenetic risk classification by Fluorescence-in-situ-hybridization analyses of CD34+ peripheral blood cells the groups differed significantly for overall and leukemia-free survival by uni- and multivariate analyses without discrepancies between treated and untreated patients. Including cytogenetic data of Fluorescence-in-situ-hybridization analyses of peripheral CD34+ blood cells (instead of bone marrow banding analysis) into the complete International Prognostic Scoring System assessment, the prognostic risk groups separated significantly for overall and leukemia-free survival. Our data show that a reliable stratification to the risk groups of the International Prognostic Scoring Systems is possible from peripheral blood in patients with missing chromosome banding analysis by using a comprehensive probe panel. This trial was registered at www.clinicaltrials.gov as #NCT01355913.
... In addition, cytogenetic subgroups and prognostic variables have recently been suggested as providing improved prognostic evaluation of clinical outcomes of primary MDS patients [18][19][20]. Although the established prognostic scoring systems are based on conventional cytogenetics, some studies showed that chromosomal abnormalities detected by fluorescence in situ hybridization (FISH) may provide prognostic information [21,22], and may be useful in supporting the clinical decision making in the selected cases, such as those with del(5q) or with del(7q) or monosomy 7 [23]. ...
Myelodysplastic syndromes (MDS) are a heterogeneous group of clonal hematopoietic stem cell disorders; they are characterized by ineffective hematopoiesis and a predilection to the development of acute myeloid leukemia (AML). For a rapid evaluation of the outcome in myelodysplastic patients non-cytogenetic prognostic scores can be used.
This study proposed to demonstrate that age and gender are important factors in the outcome of the patients diagnosed with myelodysplastic syndrome.
This study was conducted in the Department of Hematology of the Emergency University Hospital Bucharest during October 2008 and October 2012.
Male sex and age higher than 60 years are associated with high risk in the studied cases by using the Spanish prognostic score. According to Goasguen score: male sex and age, patients older than 60 years, present characteristics associated with an intermediate risk. Based on the Dusseldorf score, age over 60 years and female gender were associated with pronounced risk in the examined group. By examining the Bournemouth score in our group, we found that age > 60 years correlated with a higher frequency of risk, but no significant differences regarding the sex of patients were observed.
We concluded that age > 60 years and male gender are important predisposing factors in the survival. Abbreviations MDS = myelodysplastic syndromes, AML = acute myeloid leukemia, LDH = lactate dehydrogenase, FAB = French-American-British, WHO = World Health Organization, IPSS = International Prognostic Scoring System, ALIP = abnormal localization of immature precursors, WPSS = WHO classification-based prognostic scoring system, FISH = fluorescence in situ hybridization, del = deletion.
... La presencia de alteraciones en la FISH de pacientes con SMD y cariotipos normales se corresponde con un peor pronóstico, y se ha visto que si bien en el momento del diagnostico un 50% de los SMD tienen un cariotipo normal, la mayor sensibilidad de la FISH permite el diagnostico de lesiones sub microscópicas que pasan desapercibidas en el análisis citogenético convencional, y que juegan un papel fundamental en el estadiaje pronóstico (Rigolin GM, et al 2001) . ...
WE STUDY 4.929 MARROWS BONY GATHERED FROM 1999 TO 2009 IN CITOGENÉTICA OF THE FACULTY OF MEDICINE OF THE ULL.WE CREATE A DATABASE WITH THE 1684 WITH MIELOYD PATHOLOGIES. WE COMPARE ITS CLINICAL AND GENETIC FINDS (KARIOTYPE AND FISH), EXCLUDE THE CONGRUITY FOR HAZARD AND DEFINE THE DIAGNOSIS SUITABLE THAT WITH NEW GUIDED THERAPIES, MODIFY THE PROGNOSIS OF SOME OF THESE PATHOLOGIES.
