The Cytofluorimetric/FISH Diagnostic Approach Define a B-Cell Variant-CLL with Peculiar Clinico-Biological Features.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2077-2077
Author(s):  
Lilla Cro ◽  
Luca Baldini ◽  
Lucia Nobili ◽  
Antonino Neri ◽  
Nadia Zucal ◽  
...  
Keyword(s):  
B Cell ◽  
In Situ Hybridisation ◽  
Diagnostic Algorithm ◽  
High Serum ◽  
Diagnostic Approach ◽  
Variant Form ◽  
Fish Analysis ◽  
Fluorescence In Situ Hybridisation ◽  
Biological Features ◽  
Historical Series

Abstract In the present study, we describe the clinico-biological features of 63 cases of variant B-CLL (v-B-CLL) defined according to a previously described diagnostic algorithm for CD5+ mature B-cell leukemias, based on the combined use of Cytofluorimetric (CFM) analysis and t(11;14)(q13;q32) detection by means of fluorescence in situ hybridisation (FISH). These leukemias were characterized by SIgbright/CD23+ or CD23+/− , CD79b/CD20 bright, and negativity for t(11;14)(q13;q32). A historical series of 112 classical B-CLL was used as comparison. The mean age was 61 years (33–85) and M/F ratio 1.9. The v-B-CLL cases were significantly different from the B-CLL cases in terms of the following clinico-hematological variables: age <70 yrs, lymphocytosis <20 ×109/L (P <.0001), lymphocyte doubling time ≤ 12 months (P=.04), high serum ß2-microglobulin levels (P=.003) and presence of splenomegaly (P=.007). No differences were found in terms of sex, Hb levels, platelets count, presence of serum monoclonal component, serum LDH levels and HCV-Ab positivity. In v-B-CLL and B-CLL pts the median follow-up was respectively 56 and 84 months. The median overall survival was 80 months in v-B-CLL pts while it was not reached in B-CLL pts; 37 pts (58.7%) in the first group started chemotherapy vs 84 pts (56.4%) in the latter. The analysis of leukemic cell biological characteristics related to the prognosis was also made in part of the cases. A classic or mixed-CLL cytomorphology was seen only in 22/61 pts (36.1%). The percentage of CD38 positive cases was higher in v-B-CLL (60.3) than in B-CLL (44.6; P=.028); the IgVH mutation status, evaluated in 30/63 cases, was more frequently hypermutated in v-B-CLL group (23/30) than B-CLL (P=.03); the percentage of CFM Zap-70 positivity, evaluated in 42/63 cases, was similar in two groups (47.6 vs 57.4%, respectively). Interestingly, significant differences were found about the frequence of the recurrent chromosome alterations, evaluated in 48/63 cases, by means of FISH analysis: trisomy 12 was more frequent in v-B-CLL (37.5 vs 18.4%, p=0.02), while del13q14 was more frequent in B-CLL (14.5 vs 37.8%, P=0.007). In both groups, del 11q22.3 and del 17p13.1 were relatively rare (4 vs 8%, and 2 vs 7.7%, respectively). In conclusion, our study identifies, on the basis of a defined CFM-FISH diagnostic approach, a variant form of B-CLL that shows significant differences in terms of genetic and clinical features.

scholarly journals Reliable cell and tissue morphology-based diagnosis of endemic Burkitt lymphoma in resource-constrained settings in Ghana

2019 ◽  
Author(s):  
Cecilia Smith-Togobo ◽  
Mette Ø Pedersen ◽  
Steffen G Jensen ◽  
Babatunde Duduyemi ◽  
Richard K Gyasi ◽  
...  
Keyword(s):  
Burkitt Lymphoma ◽  
In Situ Hybridisation ◽  
B Cell Lymphoma ◽  
Diagnostic Tools ◽  
Fish Analysis ◽  
Fluorescence In Situ Hybridisation ◽  
Morphological Examination ◽  
Pathology Laboratory ◽  
Retrospective Assessment ◽  
Laboratory Investigations

Abstract Endemic Burkitt lymphoma (eBL) is an aggressive B-cell lymphoma, which is a common childhood cancer in areas with intense transmission of Plasmodium falciparum parasites. Early and accurate diagnosis is a prerequisite for successful therapy, but it optimally involves advanced laboratory investigations. These are technologically demanding, expensive, and often difficult to implement in settings where eBL is prevalent. Diagnosis is thus generally based on clinical assessment and morphological examination of tumour biopsies or fine-needle aspirates (FNAs). The purpose of the present study was to assess the accuracy of eBL diagnosis at two tertiary hospitals in Ghana. To that end, we studied FNAs from 29 eBL patients and 21 non-eBL lymphoma patients originally diagnosed in 2018. In addition, we examined 111 archival formalin-fixed and paraffin-embedded (FFPE) biopsies from Ghanaian patients originally diagnosed as eBL (N=55) or non-eBL (N=56) between 2010 and 2017. Availability-based subsets of samples were subjected to haematoxylin-eosin or Giemsa staining, C-MYC immunohistochemistry, and fluorescence in situ hybridisation (FISH) analysis of c-myc rearrangements We found a good correlation between original diagnosis and subsequent retrospective assessment, particularly for FNA samples. However, evidence of intact c-myc genes and normal C-MYC expression in samples from some patients originally diagnosed as eBL indicates that morphological assessment alone can lead to eBL over-diagnosis in our study area. In addition, several FFPE samples could not be assessed retrospectively, due to poor sample quality. Therefore, the simpler FNA method of obtaining tumour material is preferable, particularly when careful processing of biopsy specimens cannot be guaranteed. We conclude that the accuracy of eBL diagnostic tools available in Ghana is generally adequate, but could be improved by implementation of additional pathology laboratory investigations. Improved attention to adequate preservation of archival samples is recommended.

New trends in chromosomal investigation in children with cardiovascular malformations

2004 ◽  
Vol 14 (6) ◽  
pp. 622-629 ◽  
Author(s):  
Ruth Schellberg ◽  
Gesa Schwanitz ◽  
Lutz Grävinghoff ◽  
Rolf Kallenberg ◽  
Detlef Trost ◽  
...  
Keyword(s):  
Chromosomal Aberrations ◽  
In Situ Hybridisation ◽  
High Rate ◽  
Diagnostic Approach ◽  
Fluorescence In Situ Hybridisation ◽  
Cardiovascular Malformations ◽  
Genetic Investigation ◽  
Different Types ◽  
Cardiovascular Defects

We investigated a group of 376 children, seen over a period of 7 years with different types of congenital cardiovascular defects, to assess the presence of chromosomal aberrations. The diagnostic approach, achieved in 3 consecutive steps, revealed conventional chromosomal aberrations in 30 of the patients (8%) excluding trisomies 13, 18, 21. Fluorescence in situ hybridisation for microdeletions showed 51 microdeletions (15%), with 43 patients having deletions of 22q11.2, 7 patients with deletion of 7q11.23, and 1 patient with deletion of 4p16.3. In 23 patients with additional clinical abnormalities, we carried out a subtelomeric screening. This revealed, in two cases (9%), different subtelomeric aberrations, namely deletions of 1p and of 1q. Thus, subtelomeric screening proved to be a very valuable as a new diagnostic approach. Our approach to genetic investigation in three phases makes it possible to detect a high rate of pathologic karyotypes in patients with congenital cardiovascular malformations, thus guaranteeing more effective genetic counselling of the families, and a more precise prognosis for the patient.

Comparison of genomic abnormality in malignant mesothelioma by the site of origin

2014 ◽  
Vol 67 (12) ◽  
pp. 1038-1043 ◽  
Author(s):  
Maiko Takeda ◽  
Takahiko Kasai ◽  
Yasunori Enomoto ◽  
Masato Takano ◽  
Kohei Morita ◽  
...  
Keyword(s):  
Malignant Mesothelioma ◽  
In Situ Hybridisation ◽  
Homozygous Deletion ◽  
Comparative Genomic Hybridisation ◽  
Comparative Genomic ◽  
Fish Analysis ◽  
Fluorescence In Situ Hybridisation ◽  
Hybridisation Analysis ◽  
Genetic Events

AimsMalignant mesothelioma (MM) results from the accumulation of a number of acquired genetic events at the onset. In MM, the most frequent changes are losses in 9p21, 1p36, 22q12 and 14q32, and gains in 5p, 7p and 8q24 by comparative genomic hybridisation analysis. We have examined various genomic losses and gains in MM and benign mesothelial proliferation by fluorescence in situ hybridisation (FISH) analysis. 9p21 deletion was reported to be less frequent in peritoneal than in pleural MMs. This study analysed various genomic losses and gains in MM by the site of origin using FISH analysis.Materials and methodsWe performed FISH analysis using paraffin-embedded tissues from 54 cases (40 pleural and 14 peritoneal) of MMs and compared the frequency of genomic abnormality by the site of origin.Results9p21 deletion was shown in 34 of 40 cases (85%) of pleural MMs, and was less frequent in five of 14 cases (36%) of peritoneal MMs (p<0.001) by FISH analysis. By contrast, 5p15 and 7p12 amplification was more significantly frequent in peritoneal than in pleural MMs. No difference between the two sites of MM in other genes was found.Conclusions9p21 homozygous deletion assessed by FISH has been reported to be useful for differentiating MM from reactive mesothelial proliferation, but it should be noted that 9p21 deletion was less frequent in peritoneal MM. Our study suggests that the pathway of the genetic abnormality might vary between pleural and peritoneal MM.

Primary cutaneous Ewing sarcoma/primitive neuroectodermal tumour: a clinicopathological analysis of seven cases highlighting diagnostic pitfalls and the role of FISH testing in diagnosis

2009 ◽  
Vol 62 (10) ◽  
pp. 915-919 ◽  
Author(s):  
M V Shingde ◽  
M Buckland ◽  
K J Busam ◽  
S W McCarthy ◽  
J Wilmott ◽  
...  
Keyword(s):  
Differential Diagnosis ◽  
In Situ Hybridisation ◽  
Molecular Testing ◽  
Fish Analysis ◽  
Fluorescence In Situ Hybridisation ◽  
Clinicopathological Analysis ◽  
Blue Cell

Aims:To perform a clinicopathological analysis of a series of primary cutaneous Ewing sarcomas/primitive neuroectodermal tumours (ES/PNET) to highlight the pathological features, discuss the differential diagnosis, emphasise the role of molecular testing (particularly fluorescence in situ hybridisation, FISH) in diagnosis and outline the patients’ clinical course.Methods:Seven cases of primary cutaneous ES/PNET were identified from the authors’ consultation files.Results:The patients were aged 16–61 years (median 25). Five were female and two were male. Five cases involved the limbs and two the trunk. Five were initially misdiagnosed (three as carcinoma and two as melanoma). All cases were characterised histologically by sheet-like growth of small round cells with little cytoplasm and showed strong membranous staining for CD99 and positive but variable staining for FLI-1. Six patients showed an EWS rearrangement (five on FISH analysis and one on RT-PCR). All tumours were completely excised. Three patients received adjuvant chemotherapy, one of whom also received radiotherapy. Follow-up was available in all cases (range 11–57 months; median 41). No recurrences or metastases occurred.Conclusions:Although rare, primary cutaneous ES/PNET should be considered in the differential diagnosis of cutaneous “small blue cell tumours”. Immunostaining for FLI-1 and molecular testing for evidence of an EWS rearrangement are useful ancillary investigations to confirm the diagnosis. The prognosis of primary cutaneous ES/PNET appears to be more favourable than extracutaneous ES/PNET.

scholarly journals Reliable cell and tissue morphology-based diagnosis of endemic Burkitt lymphoma in resource-constrained settings in Ghana

2020 ◽  
Author(s):  
Cecilia Smith-Togobo ◽  
Mette Ø Pedersen ◽  
Steffen G Jensen ◽  
Babatunde Duduyemi ◽  
Richard K Gyasi ◽  
...  
Keyword(s):  
Burkitt Lymphoma ◽  
In Situ Hybridisation ◽  
B Cell Lymphoma ◽  
Diagnostic Tools ◽  
Fish Analysis ◽  
Fluorescence In Situ Hybridisation ◽  
Morphological Examination ◽  
Pathology Laboratory ◽  
Retrospective Assessment ◽  
Laboratory Investigations

Abstract Endemic Burkitt lymphoma (eBL) is an aggressive B-cell lymphoma, which is a common childhood cancer in areas with intense transmission of Plasmodium falciparum parasites. Early and accurate diagnosis is a prerequisite for successful therapy, but it optimally involves advanced laboratory investigations. These are technologically demanding, expensive, and often difficult to implement in settings where eBL is prevalent. Diagnosis is thus generally based on clinical assessment and morphological examination of tumour biopsies or fine-needle aspirates (FNAs). The purpose of the present study was to assess the accuracy of eBL diagnosis at two tertiary hospitals in Ghana. To that end, we studied FNAs from 29 eBL patients and 21 non-eBL lymphoma patients originally diagnosed in 2018. In addition, we examined 111 archival formalin-fixed and paraffin-embedded (FFPE) biopsies from Ghanaian patients originally diagnosed as eBL (N=55) or non-eBL (N=56) between 2010 and 2017. Availability-based subsets of samples were subjected to haematoxylin-eosin or Giemsa staining, C-MYC immunohistochemistry, and fluorescence in situ hybridisation (FISH) analysis of c-myc rearrangements We found a good correlation between original diagnosis and subsequent retrospective assessment, particularly for FNA samples. However, evidence of intact c-myc genes and normal C-MYC expression in samples from some patients originally diagnosed as eBL indicates that morphological assessment alone can lead to eBL over-diagnosis in our study area. In addition, several FFPE samples could not be assessed retrospectively, due to poor sample quality. Therefore, the simpler FNA method of obtaining tumour material is preferable, particularly when careful processing of biopsy specimens cannot be guaranteed. We conclude that the accuracy of eBL diagnostic tools available in Ghana is generally adequate, but could be improved by implementation of additional pathology laboratory investigations. Improved attention to adequate preservation of archival samples is recommended.

Prevalence and prognostic value of MYD88 and CD79B mutations in ocular adnexal large B-cell lymphoma: a reclassification of ocular adnexal large B-cell lymphoma

2021 ◽  
pp. bjophthalmol-2021-319580
Author(s):  
Marina Knudsen Kirkegaard ◽  
Marthe Minderman ◽  
Lene Dissing Sjö ◽  
Steven T Pals ◽  
Patrick R G Eriksen ◽  
...  
Keyword(s):  
B Cell ◽  
Mutational Analysis ◽  
Cell Lymphoma ◽  
In Situ Hybridisation ◽  
B Cell Lymphoma ◽  
Fluorescence In Situ Hybridisation ◽  
Large B Cell Lymphoma ◽  
Mutational Status ◽  
Large B Cell

AimsTo (1) reclassify ocular adnexal large B-cell lymphomas (OA-LBCLs) per 2016 WHO lymphoma classification and (2) determine the prevalence of MYD88 and CD79B mutations and their association with clinical parameters among OA-LBCLs.MethodsThis study is a retrospective analysis of all OA-LBCLs diagnosed in Denmark between 1980 and 2018. Medical records and tissue samples were retrieved. Thirty-four OA-LBCLs were included. Fluorescence in situ hybridisation and Epstein-Barr-encoded RNA in situ hybridisation were used for the reclassification. Mutational status was established by allele-specific PCR and confirmed by Sanger sequencing. Primary endpoints were overall survival, disease-specific survival (DSS) and progression-free survival (PFS).ResultsTwo LBCL subtypes were identified: diffuse large B-cell lymphoma (DLBCL) (27 of 32; 84%) and high-grade B-cell lymphoma (HGBL) with MYC and BCL2 and/or BCL6 rearrangements (5 of 32; 16%). cMYC/BCL2 double-expressor DLBCLs had a poorer DSS than non-double-expressor DLBCLs (5-year DSS, 25% vs 78%) (HR 0.23; 95% CI 0.06 to 0.85; p=0.014). MYD88 mutations were present in 10 (29%) of 34 lymphomas and carried a poorer PFS than wild-type cases (5-year PFS, 0% vs 43%) (HR 0.78; 95% CI 0.61 to 0.98; p=0.039). CD79B mutations were present in 3 (9%) of 34 cases.ConclusionOA-LBCL consists mainly of two subtypes: DLBCL and HGBL with MYC and BCL2 and/or BCL6 rearrangements. MYD88 mutations are important drivers of OA-LBCL. MYD88 mutations, as well as cMYC/BCL2 double-expressor DLBCL, appear to be associated with a poor prognosis. Implementing MYD88 mutational analysis in routine diagnostics may improve OA-LBCL prognostication.

Cytogenetic and fluorescence in situ hybridisation (FISH) analysis of ocular melanoma

1994 ◽  
Vol 77 (2) ◽  
pp. 182
Author(s):  
Geoffrey C. Beverstockl ◽  
Martine de Jager ◽  
Ite de Waard-Siebinga ◽  
Jeanette Kool ◽  
Paul Mollevanger ◽  
...  
Keyword(s):  
In Situ Hybridisation ◽  
Ocular Melanoma ◽  
Fish Analysis ◽  
Fluorescence In Situ Hybridisation

scholarly journals MLLrearranged acute lymphoblastic leukaemia presenting as a maxillary sinus mass with a discordant immunophenotypic profile from the bone marrow

2019 ◽  
Vol 12 (2) ◽  
pp. e227400
Author(s):  
Yunzu Michele Wang ◽  
Jun Qin Mo ◽  
Dennis John Kuo ◽  
Victor Wong
Keyword(s):  
Bone Marrow ◽  
Acute Lymphoblastic Leukaemia ◽  
B Cell ◽  
Maxillary Sinus ◽  
Lymphoblastic Leukaemia ◽  
In Situ Hybridisation ◽  
Fluorescence In Situ Hybridisation ◽  
Primary Mass ◽  
B Cell Malignancy ◽  
Cell Malignancy

We describe an unusual case of pre-B lymphoblastic leukaemia presenting with a unilateral maxillary sinus mass in which biopsies of the primary mass and the bone marrow demonstrated conflicting immunophenotyping results. The extramedullary mass was consistent with a precursor B-cell malignancy, while the bone marrow was initially reported as a possible mature B-cell malignancy. The treatments for the two are fundamentally different, which necessitated a delay in the initiation of his chemotherapy until a clear diagnosis was made. Mixed lineage leukaemia gene rearrangement was confirmed by fluorescence in situ hybridisation in both the primary mass and bone marrow, which unified the diagnosis as pre-B acute lymphoblastic leukaemia given the common cytogenetic feature.

Fluorescence in situ hybridisation analysis of bone marrow trephine biopsy specimens; an additional tool in the diagnostic armoury

2012 ◽  
Vol 66 (1) ◽  
pp. 54-57 ◽  
Author(s):  
Michael J Neat ◽  
Mufaddal T Moonim ◽  
Robert G Dunn ◽  
Helen Geoghegan ◽  
Nicola J Foot
Keyword(s):  
In Situ Hybridisation ◽  
Fish Analysis ◽  
Fluorescence In Situ Hybridisation ◽  
Trephine Biopsy ◽  
Additional Tool ◽  
Bone Marrow Trephine Biopsy ◽  
Biopsy Specimens ◽  
Wide Range ◽  
Formalin Fixed Paraffin Embedded

Fluorescence in situ hybridisation (FISH) analysis is now widely employed in the diagnosis and risk stratification of a wide range of malignant diseases. While this technique is used successfully with formalin-fixed paraffin-embedded (FFPE) sections from numerous tissue types, FISH analysis of FFPE tissue sections from trephine biopsy specimens has been less widely reported, possibly due to technical limitations relating to the decalcification protocols employed. During the last 4 years FISH analysis has been carried out successfully in 42 out of 55 (76%) consecutive trephine biopsy specimens received as part of the standard diagnostic service at our institution. Samples decalcified using EDTA-based protocols were analysed successfully in 31/31 cases (100%), whereas only 11/24 samples (46%) decalcified using formic acid-based protocols were successful. In our experience, FISH analysis of trephine biopsy specimens is a highly reproducible technique and a very useful adjunctive tool in the diagnostic armoury; however, its use in a standard diagnostic setting relies on the use of EDTA-based decalcification protocols.

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