scholarly journals The Role of Cytogenetic Methods in the Diagnosis of Haematological Diseases

2021 ◽  
Author(s):  
Amina Aščerić ◽  
Mirela Mačkić Durović ◽  
Hilada Nefić
Keyword(s):  
Chromosomal Aberrations ◽  
Cytogenetic Analysis ◽  
Chromosomal Abnormalities ◽  
Haematological Malignancies ◽  
Diagnosis And Prognosis ◽  
Conventional Cytogenetic Analysis ◽  
Frequent Use ◽  
Conventional Cytogenetics ◽  
Cytogenetic Methods

Introduction: Conventional cytogenetics by the use of standard karyotyping allows the study of numerical and structural chromosomal aberrations. Haematological malignancies include a number of cancer types that originate in the blood cells of the bone marrow or of the lymphatic system. Cytogenetic methods are traditionally used for the sake of diagnosis and prognosis of these diseases. However, with the ever more frequent use of molecular methods in the diagnostic laboratories, the importance of the conventional cytogenetic analysis in the diagnosis of haematological diseases needs to be reassessed. Aim: To evaluate the role of cytogenetic methods in the diagnosis of haematological malignancies. Materials and Methods: A retrospective analysis of cytogenetic findings of 146 patients with various haematological malignancies was performed. All of the findings were made over a period of three years at the Centre for Genetics at the Medical Faculty of the University of Sarajevo in Bosnia and Herzegovina. Microsoft Excel 2019 was used for the analysis and presentation of data in the form of tables and graphs. Results: The results of the present study showed that the use of conventional cytogenetic analysis is a good diagnostic method for 50.68% (74) of patients in whom chromosomal aberrations were detected. Conclusion: Cytogenetics remains the most comprehensive method for assessing chromosomal abnormalities due to its ability to detect clinically relevant aneuploidies and additional cytogenetic abnormalities that cannot be detected by locus-specific assays.

scholarly journals Bivariate flow karyotyping in human Philadelphia-positive chronic myelocytic leukemia

Blood ◽  
1988 ◽  
Vol 72 (1) ◽  
pp. 282-286 ◽  
Author(s):  
GJ Arkesteijn ◽  
AC Martens ◽  
A Hagenbeek
Keyword(s):  
Chromosomal Aberrations ◽  
Cytogenetic Analysis ◽  
Gradient Centrifugation ◽  
Philadelphia Chromosome ◽  
Chromosome Analysis ◽  
Chronic Myelocytic Leukemia ◽  
Translocation Event ◽  
Myelocytic Leukemia ◽  
Conventional Cytogenetic Analysis ◽  
The One

Abstract Chromosome analysis on clinical leukemia material was done by means of flow cytometry (flow karyotyping) to investigate the applicability of this technique in the detection of leukemia-associated abnormalities. Flow karyotyping was performed on blood or bone marrow samples from eight patients with chronic myelocytic leukemia (CML) after a culture period of four days and arresting the cells in metaphase during the last 16 hours. Discontinuous density gradient centrifugation proved to be essential in removing debris and dead cells from the cell suspensions. By this procedure the mitotic index increase ranged from 2 to 80 times initial values. Chromosomes were isolated and stained with two base pair-specific fluorochromes, ie, chromomycin A3 and Hoechst 33258, and run through a specially designed dual-laser beam flow cytometer. Generally, 20,000 chromosomes or more were measured. The data were computer stored in list mode. Besides the clear detection of the specific Philadelphia chromosome, trisomies and other additional chromosomal aberrations [like an i(17q)] were visualized. Quantitative analysis revealed the percentage of subclones containing a certain chromosomal anomaly. Conventional cytogenetic analysis confirmed these findings. In seven of eight cases, CML could be diagnosed on the basis of the presence of a Philadelphia chromosome in the flow karyogram. In one of these seven, the conventional cytogenetic analysis was unknown at that time. The remaining six all matched the standard cytogenetics. The one failure out of eight could be attributed to the specific stimulating conditions in the culture. Although it is impossible by this technique to determine the position of the breakpoint, the involved chromosomes in the translocation event could be identified. In some cases, low percentages of aberrations could not be detected. This study shows that CML can be diagnosed on the basis of flow karyotypic results. Additional chromosomal aberrations can be detected provided that changes in the amount of DNA per chromosome have occurred. Exact quantification of the composition of subclones in the case of mosaicism appear difficult.

Classical and Molecular Cytogenetic Abnormalities in 124 Pediatric Patients with Acute Lymphocyte Leukemia.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4419-4419
Author(s):  
Yihuan Chai ◽  
Hui Lv ◽  
Jun Lu ◽  
Peifang Xiao ◽  
Jianqing Li
Keyword(s):  
Bone Marrow ◽  
Multiplex Pcr ◽  
Cytogenetic Analysis ◽  
Chromosomal Abnormalities ◽  
Molecular Diagnostic ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Childhood All ◽  
Cytogenetic Abnormalities ◽  
Conventional Cytogenetic Analysis

Abstract In childhood acute lymphocyte leukemia (ALL), cytogenetics plays an important role in diagnosis, allocation of treatment and prognosis. On base of the conventional cytogenetic analysis, molecular methods have inproved our ability to accurately and rapidly risk-stratify patient with childhood ALL in the last few years. Our aim was to assess the demography of cytogenetic abnormalities in childhood ALL. The study sample consisted of 124 newly diagnosed ALL patients younger than 16 years, who were diagnosed at the Department of Pediatric Hematology/Oncology, Soochow University Children’s Hospital. The diagnosis and FAB subtypes of ALL was determined by Wright-Giemsa-stained bone marrow smears and cytochemicalstaining. Immunophenotyping of the bone marrow samples was performed by flow cytometry. Multiplex reverse transcription-polymerase chain reaction (RT-PCR) analysis was performed to detect the 29 most common leukemia translocations for routine molecular diagnostic hematopathology practice, and complement the information gained from conventional cytogenetic analysis. Cytogenetic analysis was successful in 112 of 124 children with ALL. Sixty-eight (60%)of them had clonal chromosomal abnormalities. Numerical imbalances consisted of hyperdipoild(>47 chromosomes, 36 cases), hypodipoild(<46 chromosomes, 14 cases), pseudodiploidy(18 cases). Chromosomal translocations were observed in 13 patients by conventional cytogenetic analysis. Three cases were found positive for t (4;11), 3 cases for t (9;22), 1 case for t (1;19) and 6 cases for other rare translocations. RT-PCR analysis detected 116 of the 124 ALL patients. Thirteen cases of TEL-AML1, 10 cases of rearrangement in the MLL gene, 4 cases of E2A-PBX1, 4 cases of E2A-HLF, 3 cases of BCR-ABL, 2 cases of TLS-ERG, 32 cases of HOX11, were detected by RT-PCR in B-lineage leukemias. SIL-TAL1 had been found in 4 of 7 of T-lineage leukemias. Sixty-eight cases of ALL show chromosomal aberrations. Multiplex PCR positivity was detected in 59(50%)of the 116 ALL patients studied. Multiplex PCR combined with chromosome analysis uncovered Chromosomal abnormalities in 95 of 124(77%) of ALL patients and supplemented each other in detecting Chromosomal abnormalities. It provides reliable evidence for the diagnosis, classification and prognosis.

The Utility of Peripheral Blood FISH in the Quantitation of BCR/ABL in CML Patients on Imatinib Mesylate: A Comparison with Bone Marrow FISH and Conventional Cytogenetics.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2942-2942
Author(s):  
Samar Issa ◽  
Duncan Holdsworth ◽  
Paul Oei ◽  
Peter J. Browett
Keyword(s):  
Bone Marrow ◽  
Imatinib Mesylate ◽  
Peripheral Blood ◽  
Cytogenetic Analysis ◽  
Fusion Gene ◽  
Chronic Phase ◽  
Conventional Cytogenetic Analysis ◽  
The Mean ◽  
Conventional Cytogenetics ◽  
Mean Differences

Abstract The hallmark of CML is the BCR/ABL fusion gene that is usually formed as a result of the t(9;22) translocation. Conventional cytogenetic analysis has been the standard method for monitoring the Philadelphia (Ph) chromosome, however evaluation of the BCR/ABL fusion gene using interphase Fluorescence in situHybridisation (FISH) on peripheral blood may allow more frequent and less invasive follow up of CML patients. The objective of this study was to compare the utility of peripheral blood FISH versus bone marrow FISH and conventional cytogenetics in patients with CML following treatment with Imatinib mesylate. 61 sets of peripheral blood and bone marrow aspirate samples from 33 Ph positive chronic phase CML patients receiving treatment with Imatinib mesylate were assessed from December 2002 to February 2004. Bone marrow samples were processed by standard cytogenetic procedures and G-banded analysis of at least 20 metaphases per sample was performed. Interphase FISH on non selected peripheral blood and bone marrow samples was carried out by scoring positive signals in 600 nuclei in each sample using BCR/ABL dual fusion or extra signal probes (Vysis). Bland and Altman plots were constructed to assess the level of agreement between the tests and the mean differences (with 95% confidence intervals) were determined. 13 of the 33 patients studied were male and 21 (64%) of the patients were analyzed at more than one timepoint. Although there was good agreement of peripheral blood FISH with bone marrow FISH and bone marrow cytogenetics in monitoring changes in the level of Ph positive cells following therapy, there were statistically significant differences in the agreement between the percentage levels of BCR/ABL positive cells between bone marrow cytogenetics and bone marrow and peripheral blood FISH. Cytogenetic analysis revealed significantly higher levels of BCR/ABL positive cells when compared to both peripheral blood FISH [9% (95% CI 4.6, 14.1), p=0.013] and bone marrow FISH [5% (95% CI 1.7, 7.8, p=0.013)]. The mean difference in the percentages of the Ph positive cells measured by bone marrow FISH exceeded the peripheral blood FISH by 5% (95% CI 1.0, 8.1, p=0.037). There was a significant relationship between the differences observed and the actual percentage level of BCR/ABL positive cells (P&lt;0.001) with most of the discrepancy being driven by tests with BCR/ABL positive cells levels between 10 and 90%. In this subgroup, the cytogenetic tests were significantly higher [mean level 34% (95% CI 26, 45), p&lt;0.001] than peripheral blood FISH and 23% (95% CI 13, 33, p&lt;0.001) higher than bone marrow FISH. These observed differences may relate to the analysis of non-dividing cells in the FISH studies, including the assessment of Ph negative T lymphocytes in the peripheral blood, and need to be considered when monitoring patients by peripheral blood FISH studies alone. Figure Figure

Clonal Cells Detected by Conventional Cytogenetic Analysis Correlates with Bone Marrow Blast Percentage and Progression Free Survival Into Acute Leukemia In Myelodysplastic Syndromes

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 2935-2935
Author(s):  
Miyoung Kim ◽  
Cha Ja She ◽  
Sang Mee Hwang ◽  
Sung-Soo Yoon ◽  
Byoung Kook Kim ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Prognostic Factors ◽  
Cytogenetic Analysis ◽  
Chromosomal Abnormalities ◽  
Progression Free Survival ◽  
Bone Marrow Blast ◽  
Free Survival ◽  
Conventional Cytogenetic Analysis ◽  
Cell Percentage ◽  
Clonal Abnormalities

Abstract Abstract 2935 Introduction: Chromosomal abnormalities not only demonstrate the clonality of the disease, but also help to predict the outcome of the patients in MDS. While IPSS suggests conventional cytogenetic analysis as a standard method in detecting clonal abnormalities in MDS, FISH is widely used due to its ability to detect minor clones and to quantitate the clonal abnormalities. In this study, we compared the results of conventional cytogenetic analysis and FISH, and investigated their different prognostic impact in MDS. Materials and Methods: The study included 129 MDS patients composed of 10 RA, 5 RARS, 15 RCMD, 39 RAEB-1, 33 RAEB-2 and 5 MDS, U (WHO classification, 2001). A standard protocol was used to perform conventional cytogenetic analysis (G-banding). Interphase FISH was performed to detect any abnormalities of chromosomes 5, 7, 8, 20 and 1. The result was analyzed as per the manufacturer¡&hibar;s instructions and recorded according to ISCN (2005) criteria. Result: The incidence of −5/5q, −7/7q, +8, −20q and +1q were 13.2%, 14.0%, 19.4%, 7.0% and 7.8%, respectively. Among them, FISH detected occult abnormalities which were not detected in conventional cytogenetic analysis in 1.6%, 3.1%, 5.4%, 1.6% and 5.4%, respectively. Overall, FISH detected occult abnormalities in 18.6% of the patients (24/129), and IPSS grouping was changed in 4.9% (6/129: 1 Low to Int-1; 4 Int-1 to Int-2; 1 Int-2 to High). The abnormalities in −5/5q, −7/7q and +1q were more common with other chromosomal abnormalities, whereas the abnormalities in +8 and −20q were more common as a sole abnormality. The quantity of clonal cells for each chromosome detected in conventional cytogenetic analysis did not correlate with the quantity of clonal cells detected in FISH. The presence of clonal cells either in conventional cytogenetic analysis or in FISH did not correlate with prognostic factors included in IPSS. However, the clonal cell percentage detected in conventional cytogenetic analysis was higher in patients with >5% bone marrow blasts than those with <5% (79.1% vs. 57.9%, p=.013). The clonal cell percentage detected in FISH did not show any relationship with IPSS factors including bone marrow blast percentage. Multivariate analysis showed the presence of clonal cells in conventional cytogenetic analysis were independent prognostic factors in predicting progression free survival into acute leukemia in this patient group (p=.038). Conclusion: FISH is advantageous in identifying cryptic cytogenetic abnormalities which could be overlooked in conventional cytogenetic analysis, and can change the IPSS risk grouping in MDS patients. The quantity of clonal cells detected in conventional cytogenetic analysis correlates with bone marrow blast percentage, whereas the quantity of clonal cells detected in FISH does not. Blasts possess higher proliferating activity than maturing hematopoietic precursors, thus they are more likely to form metaphase required in conventional cytognetic analysis. In contrast, the FISH results performed on non-dividing, interphase cells might reflect the true quantity of clonality both in blasts and maturing hematopoietic precursors. The relationship between the quantity of clonal cells and bone marrow blast percentage, and the prognostic significance of the presence of clonality suggest the novel diagnostic utility of conventional cytogenetic analysis in MDS. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Additional chromosomal abnormalities in Philadelphia positive chronic myeloid leukemia

2019 ◽  
Vol 36 (2) ◽  
Author(s):  
Sunila Tashfeen Arif ◽  
Rafia Mahmood ◽  
Saleem Ahmed Khan ◽  
Tahir Khadim
Keyword(s):  
Chronic Myeloid Leukemia ◽  
Cytogenetic Analysis ◽  
Myeloid Leukemia ◽  
Chromosomal Abnormalities ◽  
Philadelphia Chromosome ◽  
Newly Diagnosed ◽  
Cytogenetic Abnormalities ◽  
Conventional Cytogenetic Analysis ◽  
Major Route ◽  
Additional Chromosomal Abnormalities

Objective: To determine the frequency of additional chromosomal abnormalities in Philadelphia chromosome positive Chronic Myeloid Leukemia (CML) by conventional cytogenetic analysis. Methods: This descriptive cross sectional study was conducted at Armed Forces Institute of Pathology (AFIP), Rawalpindi, from January 2012 to December 2016. A total number of 528 newly diagnosed CML patients were included in the study. The subjects were tested for the presence of Philadelphia (Ph) chromosome and other additional cytogenetic abnormalities by conventional cytogenetic analysis interpreted according to International System of Human Cytogenetic Nomenclature (ISCN) criteria. Molecular analysis for BCR-ABL was also performed for each patient. The additional cytogenetic abnormalities were then classified into major route abnormalities and minor route abnormalities. Results: Out of the 528 newly diagnosed CML patients, 378 (71.6%) were males and 150 (28.4%) were females. The age of patients ranged between 18 to 74 years. Four hundred and ninety-eight (94.3%) patients showed Philadelphia chromosome on karyotyping while 30 (5.7%) were negative for the Philadelphia chromosome. On analysis of these 498 Philadelphia positive patients, additional cytogenetic aberrations were detected in 26 (4.9%) patients. Of these, 7 (1.3%) had major route abnormalities while 19 (3.6%) had minor route abnormalities. Conclusion: The frequency of additional chromosomal abnormalities in our study were not in accordance with previous local and international studies. doi: https://doi.org/10.12669/pjms.36.2.1384 How to cite this:Tashfeen S, Mahmood R, Khan SA, Khadim T. Additional chromosomal abnormalities in Philadelphia positive chronic myeloid leukemia. Pak J Med Sci. 2020;36(2):---------. doi: https://doi.org/10.12669/pjms.36.2.1384 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Optimization of modern diagnosis of chromosomal abnormalities detected in couples with reproductive dysfunction

2016 ◽  
pp. 42-44
Author(s):  
A.S. Dariy ◽  
◽  
A.A. Stepanov ◽  
S.V. Denisenko ◽  
◽  
...  
Keyword(s):  
Key Words ◽  
Cytogenetic Analysis ◽  
Chromosomal Abnormalities ◽  
Human Reproduction ◽  
Chromosomal Anomalies ◽  
Molecular Cytogenetic ◽  
Genetic Mechanisms ◽  
Balanced Translocations ◽  
Chromosomal Mutations ◽  
Cytogenetic Methods

The objective: To define the percentage of molecular-cytogenetic methods in chromosomal anomalies diagnosis among the couples who had visited Human Reproduction Problems Clinic. To define the algoritm of pregravidate service of couples using the molecular-cytogenetic results. Patients and methods. Сytogenetic analysis was performed for 1812 couples (3624 patients) who visited Human Reproduction Problems Clinic. Molecular-cytogenetic analysis was performed for 426 patients. Results. Molecular-cytogenetic methods were used in 11.75% cases. Mostly (in 78.2% cases) it was used for knowing the correlation between normal and abnormal clones in karyotype. Also it was used for identification of balanced translocations breakpoints (in 15.8% cases) and for identification of marker chromosomes (2.9% cases). Conclusions. Our results demonstrate effectiveness of molecular-cytogenetic methods for genome and chromosomal mutations analysis among couples with reproductive problems. The results were taken in account during pregravidate service of couples with reproductive problems. Knowing of genetic mechanisms of reproductive failure is importing for preconception examination. Key words: infertility, pregravidate service, chromosomal pathology, genome mutations, mosaicism, molecular-cytogenetic methods.

scholarly journals Frequency of Chromosomal Abnormalities in Products of Conception

2017 ◽  
Vol 39 (03) ◽  
pp. 110-114 ◽  
Author(s):  
Thaís Teles ◽  
Carolina Paula ◽  
Mariana Ramos ◽  
Helena Costa ◽  
Cyntia Andrade ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chromosomal Aberrations ◽  
Maternal Age ◽  
Chromosomal Abnormalities ◽  
Chain Reaction ◽  
Products Of Conception ◽  
Polymerase Chain ◽  
Conventional Cytogenetics ◽  
Quantitative Fluorescence ◽  
Abnormal Results

Purpose To describe the frequencies of chromosomal abnormalities found in abortion material, and to observe its correlation to maternal age. Methods A retrospective study was conducted based on data obtained from the databank of a medical genetics laboratory in Belo Horizonte, MG, Brazil. A total of 884 results from products of conception analysis were included, 204 of which were analyzed by cytogenetics, and 680 by molecular biology based on quantitative fluorescence polymerase chain reaction (QF-PCR). The frequency of individual chromosomal aberrations and the relationship between the presence of anomalies and maternal age were also evaluated. Results The conventional cytogenetics technique was able to detect 52% of normal and 48% of abnormal results in the analyzed material. Quantitative fluorescence polymerase chain reaction revealed 60% of normal and 40% of abnormal results from the samples evaluated by this method. The presence of trisomy 15 was detected only by cytogenetics, as it was not included in the QF-PCR routine investigation in the laboratory. A significant increase in abnormal results was observed among women aged 35 years or older compared with younger women (p = 0.02). Conclusion Chromosomal aberrations are still a major cause of spontaneous abortion, and the conventional cytogenetics technique is efficient for miscarriage material analysis, but molecular methods such as QF-PCR are adequate complementary strategies to detect the major chromosomal anomalies, leading to technical reports with reliable results.

scholarly journals Antenatal detection of chromosomal abnormalities combining QF-PCR and cytogenetic analysis

2020 ◽  
Vol 3 (1) ◽  
pp. 34-43
Author(s):  
Ana Vicic ◽  
Vedrana Skaro ◽  
Petar Projic ◽  
Petra Korac ◽  
Romana Gjergja-Juraski ◽  
...  
Keyword(s):  
Prenatal Diagnosis ◽  
Cytogenetic Analysis ◽  
Chromosomal Abnormalities ◽  
Chorionic Villus ◽  
Pcr Analysis ◽  
Chorionic Villus Sampling ◽  
Tissue Samples ◽  
Conventional Cytogenetic Analysis ◽  
Diagnostic Values ◽  
Fluorescent Polymerase Chain Reaction

Aim: To compare the diagnostic values and limitations of quantitative fluorescent polymerase chain reaction (QF-PCR) and conventional cytogenetic analysis in prenatal diagnosis of chromosomal abnormalities. Methods: A prospective study included simultaneous QF-PCR and cytogenetic analysis of 133 prenatal samples routinely obtained by amniocentesis or chorionic villus sampling (CVS). Additionally, QF-PCR analysis was performed on 14 tissue samples collected after termination of pregnancy (TOP) for which karyotyping could not be performed due to culture failure. Results: Among 133 analyzed prenatal samples, chromosomal abnormalities were diagnosed in 12 cases (9%), including 10 cases of numerical chromosomal aberrations and two cases with unbalanced structural rearrangements. Nine out of 12 chromosomal abnormalities were also detected with QF-PCR. However, all cases of major aneuploidies were successfully disclosed with QF-PCR, resulting in 100% detection rate for chromosomes 21, 18, 13, X and Y. Using a set of markers specific for chromosomes 21, 18 and 13, QF-PCR analysis of tissues collected after TOP revealed chromosomopathy in 21.4% of cases (two cases of trisomy 18 and one triploidy). A comparison of STR markers confirmed monozygosity in two monochorionic/diamniotic twin pregnancies. Conclusion: QF-PCR has been shown as a rapid and reliable method for prenatal diagnosis of the most common chromosomal aneuploidies, and as an adequate alternative to conventional karyotyping in cases where cytogenetic analysis is not possible due to failure of culturing process. However, conventional cytogenetics still presents a gold standard for the detection of structural aberrations and rare aneuploidies.

Chromosomal Aberration for Diagnosis and Prognosis of Acute Myeloid Leukemia Iraqi Patients

2021 ◽  
Vol 19 (6) ◽  
pp. 115-121
Author(s):  
Amal M. Ali ◽  
Shahlaa M Salih
Keyword(s):  
Acute Myeloid Leukemia ◽  
Chromosomal Aberrations ◽  
Myeloid Leukemia ◽  
Chromosomal Abnormalities ◽  
Gene Mutations ◽  
Newly Diagnosed ◽  
High Complexity ◽  
Diagnosis And Prognosis ◽  
Chromosomal Changes ◽  
Acute Myeloid

Background: Acute myeloid leukemia (AML) is a hematopoietic disorder in which there are too many immature blood-forming cells accumulating in the bone marrow and interfering with the production of normal blood cells. It has long been recognized that AML is a clinically heterogeneous disease characterized by a multitude of chromosomal abnormalities and gene mutations, which translate to marked differences in responses and survival following chemotherapy. This study aimed to clarify the chromosomal aberrations in early diagnosed and relapsed cases of AML. Materials and methods: Chromosomal changes were studied in thirty Iraqi patient samples diagnosed with acute myeloid leukemia were divided into 9 newly diagnosed and 13 received chemotherapy who were incomplete remission and 8 relapsed subjects. Results: Analysis of all chromosomal aberrations showed complex karyotype for most cells of relapsed AML patients compared with newly diagnosed patients. Conclusions: Chromosomal abnormalities are linked to AML development and high complexity of karyotyping for relapsed group.

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