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ABSTRACT: Using hematology analysers, white blood cell (WBC) counts and differentials (either three or five parameters) may be ascertained after Red Blood Cell (RBC) lysis and analysis using either impedance and/or optical (laser) technology. Cells or particles not destroyed by lytic agents are enumerated as WBC: abnormal particles may be observed on WBC differential scattergrams, if performed, appearing as a variable number of dots, which location may help to ascertain the nature of the abnormality. Spuriously low WBC counts are rare, mainly related to agglutination in the presence of ethylenediamine tetra-acetic acid. Cryoglobulins, lipids, insufficiently lysed RBC, erythroblasts and platelet aggregates are common situations increasing WBC counts. So far, many current high performance analysers clearly identify and enumerate erythroblasts now. In normal patients and in reactive disorders automated differential provides true and accurate results. However, failure to enumerate accurately basophilic granulocytes and monocytes is not uncommon. Using myeloperoxidase cytochemistry to ascertain differential may lead to slide review if the enzyme expression is low or absent. Low number of abnormal cells (blasts, lymphoma cells, dysplastic granulocytes) may be missed, more frequently if leukopenia is present. In many but not all instances flagging and/or an abnormal WBC differential scattergram will alert the operator. Although these flags are sensitive enough to allow the identification of several spurious counts, only the most sophisticated analysers have optimal flagging, whereas more simple ones, especially those without a WBC differential scattergram, do not demonstrate the same sensitivity for the detection of abnormal results.
Annales de biologie clinique 04/2012; 70(2):141-54. · 0.34 Impact Factor
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ABSTRACT: Several situations lead to abnormal haemoglobin measurement or to abnormal red blood cells (RBC) counts, including hyperlipemias, agglutinins and cryoglobulins, haemolysis, or elevated white blood cells (WBC) counts. Mean (red) cell volume may be also subject to spurious determination, because of agglutinins (mainly cold), high blood glucose level, natremia, anticoagulants in excess and at times technological considerations. Abnormality related to one measured parameter eventually leads to abnormal calculated RBC indices: mean cell haemoglobin content is certainly the most important RBC parameter to consider, maybe as important as flags generated by the haematology analysers (HA) themselves. In many circumstances, several of the measured parameters from cell blood counts (CBC) may be altered, and the discovery of a spurious change on one parameter frequently means that the validity of other parameters should be considered. Sensitive flags allow now the identification of several spurious counts, but only the most sophisticated HA have optimal flagging, and simpler ones, especially those without any WBC differential scattergram, do not share the same capacity to detect abnormal results. Reticulocytes are integrated into the CBC in many HA, and several situations may lead to abnormal counts, including abnormal gating, interference with intraerythrocytic particles, erythroblastosis or high WBC counts.
Annales de biologie clinique 04/2012; 70(2):155-68. · 0.34 Impact Factor
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ABSTRACT: Two pathways, hyperdiploid and nonhyperdiploid, are proposed for progression to plasma cell neoplasia. Implication of monosomy 13 (Delta13) is unclear in monoclonal gammopathy of undetermined significance (MGUS), and data on DNA content of plasma cells [DNA index (DI)] are rare.
We ascertained DI in 169 multiple myeloma (MM) and 96 MGUS patients. Interphase fluorescence in situ hybridization (FISH) coupled to cytoplasmic staining of specific Ig (cIg-FISH) was done to look for trisomies and to ascertain Delta13.
Hyperdiploidy and hypodiploidy were found in 54% and 11.5% of MGUS patients and in 59.5% and 25% of MM patients, respectively. In MGUS patients tested using probes for odd chromosomes, cIg-FISH showed association between trisomies for chromosomes 3, 7, 9, 11, or 15 and hyperdiploidy. Delta13 was found in 45.3% and 24.6% of MM and MGUS patients, respectively. Most Delta13 cases observed in MGUS were found within hyperdiploid clones, 38% versus 11% in hypodiploid cases, in sharp contrast with the occurrence of Delta13 in MM patients, 31.9% and 76.3%, respectively. That peculiar distribution of Delta13 according to DI persisted with other thresholds used to ascertain hyperdiploidy, such as DI >or= 1.05. A strong relationship between IgA peak and hypodiploidy (P = 0.007) was only observed in MM, whereas lambda light chain was significantly associated with hypodiploidy in MGUS (P = 0.001) and MM (P = 0.05). Hyperdiploidy shows similar pattern in MGUS and MM.
This fits well a hyperdiploid pathway leading to MM after a preceding MGUS stage. Yet-to-be-determined secondary event(s) needs to occur for the transition to MM, unrelated to changes in chromosome number or to loss of chromosome 13. In contrast, the "nonhyperdiploid" pathway needs to be clarified further because hypodiploidy is less common in MGUS than in MM and Delta13 is rare in hypodiploid MGUS patients compared with hypodiploid MM patients.
Clinical Cancer Research 11/2007; 13(20):6026-31. · 7.74 Impact Factor
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Xavier Leleu,
Franck Genevieve,
Romain Guieze,
Alain Duhamel,
Joris Andrieux,
Celine Berthon, Alban Godon,
Sophie Prat-Lesaffre,
Stephane Depil,
Jean-Luc Lai,
Thierry Facon,
Marc Zandecki
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ABSTRACT: Morphological changes of plasma cells (PC) are common in multiple myeloma (MM). Loss of round or oval nuclear shape has been related to cell malignancy in human, and we looked for the occurrence of such morphological change on PC from bone marrow (BM) smears in a retrospective series of 169 MM patients at diagnosis. Nuclear shape changes of PC differed according to the patients (notch, dumb-bell, folded or monocytoid appearance), even in the same patient; all subtypes were pooled and defined as PC with irregular nuclear shape (PCIN). A significant number of PCIN (>/=5% of all BMPC) was found at diagnosis in 20.7%. Median survival was of 22 months for patients with >/=5% PCIN, and 41 months for others (p=0.0001). Significant relationship was observed with prognostic parameters related intrinsic malignancy of the tumour process but not with beta-2-microglobulin (b2m). A clear-cut relationship was found also between PCIN and hypodiploidy (p=0.0001), but not with deletion of chromosome 13. This study emphasises the relationship between PCIN, an easy-to-ascertain marker of intrinsic malignancy of the tumour process, and adverse prognosis.
Leukemia Research 06/2005; 29(6):665-71. · 2.92 Impact Factor
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ABSTRACT: Cytogenetic studies in lymphomas classically require fresh or frozen tissue, whereas in many instances only paraffin-embedded biopsies are available. We applied an interphase FISH assay on nuclei extracted from thick paraffin sections to determine accuracy of molecular cytogenetics in such samples. Twenty-three lymphoma samples and 4 reactive lymph nodes were tested with various commercially available DNA probes, and hybridization patterns were compared with those obtained on frozen nuclei counterparts. Successful hybridization with all probes tested was observed for 23/27 (85%) paraffin-embedded tissues and for all (100%) frozen samples, and cut-off levels defining positivity were superimposable for both situations. Chromosome changes were detected in the same way, without any false-positive or false-negative cases. Hybridization signals observed on dewaxed samples were either those classically expected to define the relevant chromosome change or were atypical: all atypical changes could be demonstrated also into nuclei from the frozen counterpart. Moreover, all typical and atypical chromosome changes observed on frozen nuclei were also detected in paraffin-embedded tissues. Our study shows that our interphase FISH assay performed on paraffin-embedded samples is a valuable alternate to conventional methods to ascertain diagnosis of lymphomas as to include patients into therapeutic trials.
Diagnostic Molecular Pathology 07/2004; 13(2):97-104. · 2.26 Impact Factor
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ABSTRACT: Hematology analysers provide now quick, accurate, and reproducible cell blood counts. However, depending on detection methods, spurious counts may occur. If undetected, such spurious counts may lead to inappropriate medical care and to unneeded explorations. Focusing first on platelet counts, situations leading to spurious decrease include several preanalytical considerations, the major one corresponding to EDTA-induced platelet aggregation and to platelet satellitism around polymorphs. In other instances, related to the presence of small particles mimicking platelets, including fragmented red blood cells, lipids, cryoglobulins, fibrin strands, or cytoplasmic fragments of leukocytes, spurious elevation of platelet count may occur. According to the analyser and to the methods used for the determination of the cell blood count, flags or messages related to these spurious changes differ. For each spurious change, the authors describe the mechanism leading to the anomaly, the way the analysers generate flags, and what should be done to provide accurate results.
Annales de biologie clinique 68(4):393-407. · 0.34 Impact Factor
