Automated Sysmex KX-21 hematology analyzer (Sysmex Corporation, Kobe, Japan) was used for the determination of red blood cells (RBC), hemoglobin (HGB), hematocrit (HCT), Mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red cell distribution width (RCDW), white blood cell (WBC), monocytes (MO), lymphocytes (LY), platelets (PLT). Platelets crit (PCT), platelet density width (PDW), mean platelet volume (MPV) and granulocytes (GR).
Kx 21 hematology analyzer
Manufactured by Sysmex
Sourced in Japan, Canada
The Sysmex KX-21 is a compact, automated hematology analyzer designed for routine blood cell analysis. The core function of the KX-21 is to provide accurate and reliable measurements of various blood cell parameters, including red blood cells, white blood cells, and platelets.
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22 protocols using kx 21 hematology analyzer
1
Automated Hematology Analysis Protocol
2
Blood parameter analysis protocol
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After 6 hours of treatment with LPS and D-galactosamine, blood was collected through retro-orbital route into sterile 1.5 ml Eppendorf tube. The blood was immediately processed for analyzing blood parameters using a SysmexKX-21 Hematology Analyzer (Sysmex Corporation).
3
Hematological Indices in Sudanese Population
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The hematological indices of hemoglobin (Hb) concentration, mean corpuscular volume (MCV), mean corpuscular Hb (MCH), mean corpuscular Hb concentration (MCHC), red cell distribution width (RDW), RBC count, total and differential WBC counts and platelet count were measured using Sysmex KX-21 Hematology Analyzer (Sysmex Corporation, Kobe, Japan) at the Hematology Laboratory of the Clinical Laboratory Unit at Kosti Teaching Hospital. In this reference laboratory, the hematology analyzer is regularly checked for performance to ensure that all hematological parameters of control samples are within the precision specifications. Specific, high-quality diluents were used and SOPs were followed in all measurements of hematological indices. Internal quality control is performed twice a day in the morning and afternoon before sample testing. Moreover, external quality assessment is done three times a year.
According to the criteria of WHO, anemia was defined as an Hb concentration < 11.5 g/dL in children, < 12.0 g/dL in non-pregnant women and < 13.0 g/dL in men [21 ]. Thrombocytopenia was defined as a platelet count < 150.0 × 109/L, while leucopenia was defined as a total WBC count < 4.0 × 109/L [22 ]. MCV < 80 femtoliters (fL), MCH < 27 picograms (pg), MCHC < 32 g/dL were considered low, while RDW > 14.5% was considered high [22 ].
According to the criteria of WHO, anemia was defined as an Hb concentration < 11.5 g/dL in children, < 12.0 g/dL in non-pregnant women and < 13.0 g/dL in men [21 ]. Thrombocytopenia was defined as a platelet count < 150.0 × 109/L, while leucopenia was defined as a total WBC count < 4.0 × 109/L [22 ]. MCV < 80 femtoliters (fL), MCH < 27 picograms (pg), MCHC < 32 g/dL were considered low, while RDW > 14.5% was considered high [22 ].
4
Socio-demographic and Clinical Data Collection
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Socio-demographic characteristics and clinical data were collected by trained nurses using a pre-tested semi-structured questionnaire. The data were collected by interview and review of the medical record. The baseline data were collected from the patients’ medical charts. Trained laboratory technologists collected a volume of 3–5 mL EDTA anticoagulated whole blood from each study participant and performed the laboratory analysis. Hematological parameters were determined using Sysmex kx-21 hematology analyzer (Sysmex Corporation, China), and CD4+ T cells were determined using the BD FACSCOUNT system (Becton Dickinson and Company, California, USA). The manufacturer instructions were strictly followed for each parameter.
5
Leukocyte Subset Isolation and DNA Extraction
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Individual leukocyte subsets were isolated from PB or BM via staining for specific cell surface antigens including CD4+, CD8+, CD19+, CD56+, CD33+, and CD15+ as part of routine diagnostics in HSCT recipients at the St. Anna Children’s Hospital, Vienna, Austria. Cell sorting was performed on a FACSAria instrument (BD Biosciences) using the FACSDiVa software, as described previously (Fritsch et al., 1999 (link)). The purity of individual leukocyte fractions was >98%. Generally, 4,000 cells from each leukocyte subset were isolated for subsequent analysis. For DNA isolation from plasma, PB samples were centrifuged at 100 g for 10 min, and the cell pellets were discarded. Erythrocytes derived from PB specimens of patients from the control group were obtained upon cell sedimentation. The platelet concentrates were centrifuged at 1,600 g for 30 min and resuspended in PBS for DNA isolation, as described below. Erythrocyte and platelet numbers were determined by a Sysmex Kx21 Hematology Analyzer (Sysmex).
6
Sickle Cell Disease Genotyping Protocol
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The genotype diagnosis was based on a single heavy band in the position of HbS on alkali hemoglobin electrophoresis supported by a positive solubility test. Family studies were not routinely performed, but in 7 (1.2%), a parent had an AA phenotype consistent with sickle cell-beta + thalassemia and the remaining 647 patients were assumed to have SS disease. Although not confirmed by structural studies, the cases of sickle cell-beta + thalassemia were likely to have the IVS1-5G>C mutation common in this population[7 (link)] and associated with very low levels of HbA; the genotypes have been combined for the present report. A venipuncture sample was collected in ethylenediaminetetraacetic acid (EDTA) for daily complete blood counts or more often if a fall in hemoglobin is suspected. Red cell indices were measured electronically (KX 21 hematology analyzer, Sysmex, Japan). Thin and thick blood smears were made directly from a blood drop from the syringe before the rest of the sample was collected into EDTA, stained by Giemsa stain and examined for malaria parasites under a high power oil immersion lens by the same observer (B.P.).
7
Cardiovascular Risk Factor Assessment
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Demographic and baseline data were collected by questionnaire at patient interviews and by review of medical records. Medical records were analyzed for the patients’ CAD risk factor profiles. Baseline data included gender, age, body mass index, history of smoking or drinking, family history of CAD, medical history of hypertension, type II diabetes mellitus, cranial vascular accidents, systolic and diastolic pressure, and the presence of other diseases. A TBA-120FR auto-biochemical analyzer (Toshiba, Japan) and Sysmex kx-21 hematology analyzer (Sysmex, Japan) were used to measure fasting blood sugar, glycated hemoglobin (HbA1c), creatine kinase and CKMB, troponin I (normal range 0.05–0.40 ng/mL), triglycerides (TG), total cholesterol (TC), non-high-density lipoprotein cholesterol (NHDL-C), HDL-C, and fibrinogen (Fib) in the morning after hospital admission, following an overnight fast.
8
Plasma Hemoglobin and Anemia Assessment
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Study outcomes included plasma hemoglobin concentrations (g/dL, measured by Sysmex KX21 Hematology Analyzer, Japan) and presence of anemia44 (link). Anemia was defined by hemoglobin concentrations of less than 12.0 g/dL for men and less than 11.0 g/dL for women51 (link).
9
Fasting Metabolic and Inflammatory Profile
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Subsequent to and overnight fasting, the serum levels of total cholesterol, TG, HDL, low-density lipoprotein (LDL), liver enzymes, creatinine, blood urine nitrogen (BUN), and FBS were determined exerting clinical biochemistry autoanalyzer BT3000 Plus (Biotecnica Instruments SPA, Italy) using commercial reagents (Pars Azmoon, Iran). Blood leukocytes were enumerated using Sysmex KX-21 hematology analyzer. ESR was measured using the automated kineticphotometric method (Automatic ESR analyzer, XC-A30, Caretium Medical Instruments, China). Inflammatory indices like C-reactive protein (CRP) were measured by the Enzyme linked immunosorbent assay (ELISA) using commercial kits reagents (Pars Azmoon, Iran) and microplate reader (Stat Fax 4200, Awareness Technology Inc., UAS) Blood oxygen saturation was measured using fingertip Pulse Oximeter (Beurer PO 80, France).
10
Comprehensive Hematological Analysis
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Hematological analysis (Sysmex KX-21 Hematology Analyzer; Mundelein, IL, USA) was performed using standard techniques for red blood cell count (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red blood cell distribution width (RDW), platelet count (PLT), white blood cell count (WBC), lymphocyte percentage (LYM%), lymphocyte count (LYM#), platelet distribution width (PDW), mean platelet volume (MPV), and platelet large cell ratio (P-LCR) concentrations.
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