Sysmex Corporation Research Articles

Road to automated urinalysis analyzer: Validation of Sysmex UN-3000 to supplant manual urinalysis

Abstract Introduction/Objective Traditionally, urinalysis encompasses the chemical analysis of urine by dipstick and manual microscopy for formed elements. High specimen volume and varying subjectivity among individuals performing microscopic analysis make the practice both imprecise and inefficient. Implementation of Sysmex UN- Series™Automated Urinalysis (Sysmex Corporation, Kobe, Japan) fulfilled our need for automation, addressing both issues at hand. Automated urinalysis offers a solution for laboratories to meet the growing demand for faster result turn around times(TAT) with ever increasing specimen volumes. We expect greater sensitivity and reproducibility in terms of comparison to manual microscopy, as well as quicker TAT with the change in methodology. Methods/Case Report Using both unpreserved and preserved urine tubes, a macroscopic comparative study between Clinitek Advantus(manual) and Clinitek Novus (automated) methods was carried out on 141 urines with greater than 60 abnormal patients. Similarly, a microscopic study between the laboratory’s manual microscopy procedure and the Sysmex UF-5000 was carried out on 112 urines with greater than 60 abnormal patients. RBCs, WBCs, Epithelial cells were evaluated under high power field(40X); casts and crystals were evaluated under lower power field(10X). The Reference Range Interval for both chemical and microscopic was verified using 44 normal patient specimens. The quantified values are converted to high and low power field (hpf/lpf), analogous to the current system in the laboratory. Results (if a Case Study enter NA) Most parameters had >90% agreement between manual and automated method. Parameters falling below 90% were checked to see if discrepancies fell within 1 degree. Automated system is more sensitive than manual and our average TAT for urinalysis dropped 50%. Conclusion The automated urinalysis technology enabled us to focus only on samples requiring intervention based on pathological criteria, resulting in reduced overall TAT and workload burden, and improved consistency in reporting across specimens.

Optimizing the Diagnosis of Microcytic Hypochromic Anemia: A Comparative Evaluation of Erythrocyte and Reticulocyte Parameters.

Background Anemia is a widespread, worldwide hematological disorder, with iron deficiency anemia (IDA) and thalassemia trait (TT) being two frequently encountered forms. For effective management, it is vital to distinguish between these two conditions. Existing diagnostic methods have some limitations due to their inaccessibility and high cost. This study aims to evaluate novel hematological indices for distinguishing IDA from TT, offering potential improvements in diagnostic accuracy and patient care. Objective To compare the efficacy of novel hematological indices in differentiating IDA from TT, and to assess their sensitivity and specificity in clinical practice. Methods This cross-sectional observational study included 80 anemia patients (40 diagnosed cases of IDA and 40 of TT each). Hematological parameters were measured using the Sysmex XN-1000 analyzer (Sysmex Corporation, Kobe, Japan), and statistical analysis was conducted using the Mann-Whitney test, Pearson correlation coefficient (r), and correlation matrix. Results Significant differences were observed between IDA and TT patients in the indices studied. The Green and King Index demonstrated high diagnostic accuracy, with a sensitivity of 90% and specificity of 82%. The red cell distribution width (RDW) Index exhibited similar diagnostic performance, with a sensitivity of 90% and specificity of 77%. The England and Fraser Index showed a sensitivity of 85% and specificity of 80%, with a large effect size (r = -0.60). The correlation matrix revealed strong associations between key parameters, including a negative correlation between hemoglobin and RDWIndex (r = -0.61) and a positive correlation between mean corpuscular volume (MCV) and Shine and Lal Index (r = 0.82). On the other hand, our study found that indices like Shine and Lal and Srivastava had limited diagnostic value, with smaller effect sizes (r = 0.28 and r = -0.005, respectively). Conclusion Hematological indices, such as the Green and King, RDW, and England and Fraser indices, show strong potential for differentiating IDA from TT, with high sensitivity, specificity, and large effect sizes. The correlation matrix further supports the diagnostic utility of these indices. These tools can enhance diagnostic precision in resource-limited settings and improve patient outcomes.

The Significance of Platelet Indices in the Evaluation of Thrombocytopenia.

Thrombocytopenia, characterized by a low platelet count, poses a risk of abnormal bleeding. Traditional diagnostic methods focus on platelet count alone, but emerging evidence suggests that platelet indices like mean platelet volume (MPV) and platelet distribution width (PDW) could provide valuable insights. This study aims to investigate the role of platelet indices in thrombocytopenia assessment, exploring their potential as additional diagnostic and prognostic markers. Over a five-month period, this prospective study enrolled 80 adult patients with thrombocytopenia. Platelet indices were measured using an automated blood cell analyzer (SYSMEX XN 1000; Sysmex Corporation, Kobe, Japan), and statistical analyses were performed using the StatisticalPackage for the Social Sciences (IBM SPSS Statistics for Windows, IBMCorp., Version 23.0, Armonk,NY). The study participants showed significant age differences between hypoproductive and hyperproductive thrombocytopenia groups but no significant gender-based disparities. While platelet count and plateletcrit (PCT) didn't differ significantly between groups, individuals with hyperdestructive thrombocytopenia had higher MPV values. Platelet indices varied across clinical conditions, highlighting their potential diagnostic and prognostic value. Platelet indices like MPV, along with platelet count and PCT, offer insights into thrombocytopenia causes and prognosis. Further research is needed to confirm these findings, but integrating platelet indices into clinical practice could inform treatment decisions and reduce unnecessary procedures like bone marrow biopsies and imaging studies.

Concordance between platelet counts by impedance and optical technique during microcytic anemia: towards a threshold value of the mean corpuscular volume.

Platelet count is crucial for clinical decision. In cases of microcytosis, platelet count based on impedance technique (PLT-I) may overestimate platelet count. To compare PLT-I with platelet count using the optical technique (PLT-O) and establish a Mean Corpuscular Volume (MCV) threshold for considering PLT-O. A prospective analytical study conducted over two months involved blood samples collected in standard K2 EDTA tubes for complete blood count analysis, revealing microcytosis (MCV<80 fL). PLT-O analysis in channel-Ret mode was performed using the Sysmex-XN1000 (Sysmex Corporation, Kobe, Japan). Percentage of fragmented red cells (FRC%) and percentage of microcytic red cells (Micro-R%) were recorded. Blood smears stained with May-Grünwald-Giemsa were examined for potential interfering particles. A strong correlation was observed between the two techniques for all platelet values as well as for PLT <150 x 109/L (correlation coefficient r = 0.971, 95% CI: [0.956-0.982]; P<10-3 and r = 0.90, 95% CI: [0.79-0.95]; P< 10-3). The Bland-Altman plot revealed a bias of 16.53 x 109/L between the two methods, with agreement limits between -55.8 and 88.8 x 109/L. A threshold MCV value indicating the use of the optical method, with a cut-off at 72.9fL, demonstrated promising performance consistent with literature findings. However, less favorable performance was observed with Micro-R%. Impedance could be employed in routine practice. However, for MCV<72.9 fL or in the presence of schizocytes, the hemogram validation procedure may incorporate the use of PLT-O.

Exploring the Clinical and Hematological Characteristics of Beta-Thalassemia Trait: A Comprehensive Analysis in a Tertiary Care Hospital Setting.

Beta-thalassemia is one of the most common inherited hematological diseases caused by more than 350 mutations in the β-globingene (HBB). Beta-thalassemia carrier or trait is associated with defects in one allele of the HBB gene. The majority of beta-thalassemia trait cases remain concealed in society and remain unnoticed as they are mostly asymptomatic or present with mild symptoms of anemia. There is a 25% chance of having children with beta-thalassemia major and a 50% chance of having carrier babies when two people with beta-thalassemia trait are married. Hence, it is important to identify the individuals with beta-thalassemia trait and provide counseling to understand the risks of pregnancy and its outcome. Aim To study the identification of beta-thalassemia trait cases along with their clinical findings and hematological correlation. Materials and methods Study Design This was a retrospective study conducted at Saveetha Medical College and Hospitalfor a period of four years from January 2020 to December 2023. Inclusion Criteria Age group more than 18 years, antenatal mother, cases of anemia who were refractory to iron treatment, and screening of family members in the positive cases of beta-thalassemia trait. Exclusion Criteria History of blood transfusion within three months was excluded. Data Collection A total number of 837 cases were screened to rule out the presence of beta-thalassemia trait/hemoglobin (Hb) variants. A 2 mL of intravenous blood samples were collected in an ethylene diamine tetraacetic acid (EDTA) vacutainer tube and processed in a Sysmex XN 1000 (Hyogo, Japan: Sysmex Corporation)automated hematology analyzer. The hematological parameters were analyzed. Statistical Analysis The study included both descriptive and analytical characteristics. Mean and standard deviation (SD) were calculated for all the hematological parameters. Beta-thalassemia trait was diagnosed with an HbA2 level of more than 4.0% through high-performance liquid chromatography (HPLC) analysis. Results Among the 837 samples studied for HPLC screening, 74 (8.8%) cases were found to have beta-thalassemia trait. The age group included was from 18 years to 56 years. Of 74 cases studied, 32(43%) were females and 42(57%) were males. Among the 74 cases studied, the Mentzer index <13 was seen in 58 (78%) cases and the Mentzer index >13 was seen in 16 cases (22%). Thirty-four cases (46%) of beta-thalassemia traits presented to the hospital with a history of fever for evaluation and antenatal screening accounted for 19 cases (26%). The mean red blood cell (RBC) count was 5.5 million/cu.mm; mean corpuscular volume (MCV) was 63.8 fL; mean corpuscular hemoglobin (MCH) was 19.6 pg; red cell distribution width coefficient of variation (RDW-CV) was 17.8%. Among the 74 cases studied, 37(46%) cases had an Hb of more than 11 g/dL, 22 cases had mild anemia, 12 cases had moderate anemia, and three cases had severe anemia. Conclusion This study concluded that regular monitoring of the Mentzer index along with HPLC analysis is an effective approach in identifying beta-thalassemia trait cases and further providing genetic counseling among the couples that will help in reducing high-risk pregnancy and the birth of a child with thalassemia major.

Determination of Platelet Count and Platelet Indices among Sudanese Patients with Chronic Kidney Disease

Background: Patients with chronic kidney disease (CKD) are at a considerably higher risk of thrombotic and hemorrhagic challenges. Platelet function studies in CKD are contradictory, ranging from decreased platelet function to normal or even increased platelet reaction. Our study aims to evaluate the change in platelet count (PC) and platelet indices (platelet distribution width [PDW], platelet large cell ratio [PLCR], and mean platelet volume [MPV]) among Sudanese patients with CKD.Methods and Results: This case-control study was conducted from February to August 2014 at East Nile Hospital, Khartoum, Sudan. The study involved 75 patients diagnosed with CKD (mean age 52.43±18.4 years) and 75 healthy individuals (mean age 50.3±14 years) as a control group. PC, PDW, PLCR, and MPV tests were conducted using a Sysmex KX-21N Automated Hematology Analyzer (Sysmex Corporation, Japan), and creatinine level was measured by Roche/Hitachi Cobas C311 analyzer (Basel Switzerland). The creatinine level was significantly higher in CKD patients than in the control group: 7.91±4.8 mg/dL and 1.4±1.3mg/dL, respectively (P=0.000). We found an insignificant difference between CKD patients and the control group in terms of PC, MPV, PDW, and PLCR; an insignificant difference in PC and all PI between CKD patients with creatinine levels &lt;6 mg/dL and &gt;6mg/dL; and an insignificant difference in PC and all PI between the group with CKD duration &lt;2 years and &gt;2 years and between CKD patients on hemodialysis and without hemodialysis. Conclusion: This study found no difference in PC and studied platelet indices in CKD patients, compared to the control group and no difference in PC and PI (MPV, PDW, and PLCR) in patients on hemodialysis versus patients not on hemodialysis.

Diagnostic Utility of Reticulocyte Hemoglobin Equivalent Parameter in the Evaluation of Microcytic Hypochromic Anemia – Our Experience from Northeast India

Background: Reticulocytes are red blood cell precursors with an average lifespan of 1–2 days. Reticulocyte hemoglobin equivalent (Ret-He) is an early marker of iron deficiency (ID) erythropoiesis and reflects real-time information regarding the synthesis of young erythrocytes in the bone marrow. The objective of this study is to determine the diagnostic utility of Ret-He in patients having microcytic hypochromic anemia in comparison with serum ferritin and iron studies. Objectives: The objective of this study is to determine the diagnostic utility of the Ret-He parameter in patients having microcytic hypochromic anemia in comparison with the serum ferritin, iron studies and its role in routine hematological screening for nutritional deficiency anemia like ID. Design and Settings: The design involves observational study. Materials and Methods: A hospital-based observational study was carried out in a referral hospital. Hematological parameters were processed using Sysmex XN1000 (Sysmex Corporation, Kobe, Japan) analyzer and were compared with Serum iron studies using biochemistry analyzer (VITROS® 250 Chemistry System) from 201 participants who presented with microcytic hypochromic anemia. Main Outcome and Measures: Relationship of Ret-He parameter and its diagnostic utility for screening of iron-deficient states. Sample Size: The sample size was 201. Results: When serum ferritin is taken as “the gold standard” to detect ID, with Ret-He cutoff of 27.15 pg/cell, we found a statistically significant positive correlation between Ret-He and serum ferritin (P &lt; 0.001). We found a sensitivity of 57.37% and specificity of 75.95% with area under the curve of 0.681, positive predictive value of 100%, negative predictive value of 3.8%, and accuracy of 62.19% for Ret-He in detecting ID anemia (IDA). Also found a statistically significant negative correlation between Ret-He and total iron binding capacity (P &lt; 0.001). There was no statistical correlation between Ret-He and serum iron levels in our study. Conclusions: The present study suggests Ret-He is one of the better and more reliable hematological parameters indicating ID and, when used along with biochemical parameters like serum ferritin, can give valuable inputs in a better screening and diagnosis of IDA; hence proper treatment is possible. Limitations: The multicentric study is required to standardize Ret-He reference values, and follow-up to therapy of the subjects was not done. Additional hemoglobin variant analysis data would have been favorable to the study.

ICSH review of internal quality control policy for blood cell counters.

This paper is a report of an ICSH review of policies and practices for internal quality control (IQC) policy for haematology cell counters among regulatory bodies, cell counter manufacturers and diagnostic laboratories. It includes a discussion of the study findings and links to separate ICSH guidance for such policies and practices. The application of internal quality control (IQC) methods is an essential pre-requisite for all clinical laboratory testing including the blood count (Full Blood Count, FBC, or Complete Blood Count, CBC). The ICSH has gathered information regarding the current state of practice through review of published guidance from regulatory bodies, a questionnaire to six major cell counter manufacturers (Abbott Diagnostics, Beckman Coulter, Horiba Medical Diagnostic Instruments & Systems, Mindray Medical International, Siemens Healthcare Diagnostics and Sysmex Corporation) and a survey issued to 191 diagnostic laboratories in four countries (China, Republic of Ireland, Spain and the United Kingdom) on their IQC practice and approach to use of commercial IQC materials. This has revealed diversity both in guidance and in practice around the world. There is diversity in guidance from regulatory organizations in regard to IQC methods each recommends, clinical levels to use and frequency to run commercial controls, and finally recommended sources of commercial controls. The diversity in practice among clinical laboratories spans the areas of IQC methods used, derivation of target values and action limits used with control materials, and frequency of running commercial controls materials. These findings and their implications for IQC Practice are discussed in this paper. They are used to inform a separate guidance document, which proposes a harmonized approach to address the issues faced by diagnostic laboratories.

Verification of a Benchtop Hematology Analyzer with a 5-Part Differential Count: There is Nothing Wrong with Being Small.

The benchtop ADVIA 560 AL hematology analyzer (Siemens Healthineers Tarrytown, NY, USA) offers a small footprint and ease of operation making it suitable for satellite laboratories and intensive care units. A verification study of this analyzer was performed. Between- and intra-run precision, carry-over, linearity, and throughput were evaluated on the ADVIA 560 AL. Accuracy was assessed on 94 patient samples by comparing the results obtained on the ADVIA 560 AL to the results on the reference Sysmex XN1000 analyzer (Sysmex Corporation, Kobe, Japan). The ADVIA 560 AL showed acceptable imprecision on control material and minimal bias in comparison to the XN 1000 on patient samples with a throughput of 60 samples per hour. The percentage carryover was not significant and the linearity was within acceptable limits. The ADVIA 560 AL bench-top analyzer is suitable for acute care centers and satellite laboratories owing to its small footprint, ease of use, and reproducible and accurate results.

Evaluation of Sysmex UF-5000 flow cytometer flag BACT-info for Gram discrimination in urinary tract infection

Urine culture as a gold standard for the diagnosis of urinary tract infection (UTI) involves a considerable workload in Clinical Microbiology Departments, due to the high number of samples received that will ultimately be negative. Therefore, it is necessary to use screening systems that also reduce the turnaround time for UTI diagnosis. The new flow cytometer UF-5000 (Sysmex Corporation) is able to differentiate between Gram-negative and Gram-positive bacteria using the BACT-info parameter according to manufacturer. The aim of our study was to evaluate the gram discrimination ability of the UF-5000 cytometer. A prospective study with 449 urine samples collected consecutively was conducted, in the period 7/3/2022-27/5/2022, in which the BACT-info flag was compared with urine culture as the reference method. The sensitivity obtained for both Gram-negative and Gram-positive bacteria was above 95%. However, for Gram-positive bacteria, the moderate Kappa index (0.49) and the low positive predictive value (37.1%) indicated that the correlation between BACT-info flag and urine culture was not acceptable and should not be reported to the requesting clinician. Implementation of the third generation UF-5000 cytometer represents a significant advance in the aetiological orientation of UTIs caused by Gram-negative bacteria. Reporting the Gram morphology in the urine samples reduces the response time in the microbiological diagnosis of UTI, which would have an impact on the reduction and optimisation of empirical treatment, and thus on the generation of antimicrobial resistance.

Differentiation of Acquired Hemophilia a from Lupus Anticoagulant Using Global Coagulation Assays and Magnesium-Dependent Activated Partial Thromboplastin Time Assay

CONCLUSION Rapid and reliable differentiation between bleeding and thrombotic tendency in cases with prolonged activated partial thromboplastin time (APTT) remains challenging in clinical practice, in particular, distinguishing cases positive for coagulation factor inhibitor or lupus anticoagulant (LA). To differentiate acquired hemophilia A (AHA) from LA using three global coagulation assays: rotational thromboelastometry (ROTEM), thrombin generation assay (TGA), and clot waveform analysis (CWA); also to validate the effectiveness of magnesium-dependent APTT (APTT-Mg) assay. This method uses APTT-Mg and normal APTT using Ca to screen for LA, and APTT-Mg/APTT ratio is smaller in LA. We included patients with prolonged APTT without congenital coagulation factor deficiency who were referred to our department between November 2019 and September 2021. APTT prolongation was defined as &amp;gt;35 s (normal range: 25-35 s) measured routinely with Coagpia APTT-N (Sekisui Medical). The AHA group had a confirmed diagnosis and underwent treatment of AHA. The LA group was divided into LA1 (possible LA) and LA2 (definite LA). LA1 was defined by any of the following conditions: (1) index of circulation anticoagulant ≥15 in the cross-mixing test performed with Coagpia APTT-N; (2) positive dilute Russell's viper venom time (DRVVT) phospholipid neutralization method using Gladipore LA (Medical &amp; Biological Laboratories); (3) positive APTT phospholipid neutralization test using Hemosil SCT (Instrumentation Laboratory); (4) positive for anti-CL antibody; (5) positive for anti-CLβ2GPI antibody; or (6) positive for phosphatidylserine-dependent anti-PT antibody. LA2 was defined by index of circulation anticoagulant &amp;gt;12.4 in the cross-mixing test performed by Thrombocheck APTT-SLA (Sysmex Corporation), or item (2) or (3) above, in accordance with criteria of LA in the International Society of Thrombosis and Haemostasis. LA1 and LA2 did not overlap, because cases that did not meet the conditions of LA2 were treated as LA1, and global coagulation assays were performed in all LA cases. In the global coagulation assays, 12 healthy volunteer samples were tested and regarded as the normal goroup. For the APTT-Mg assay, equal volumes of 25 mmol MgCl 2 and 25 mmol CaCl 2 were used instead of the normally used CaCl 2, and Thrombocheck APTT-SLA was used to measure APTT-Mg, which was then divided by the normal APTT concentration measured with the same APTT reagent. Ten patients in the AHA, 12 in the LA1, and 32 in the LA2 groups were eligible. ROTEM showed that clotting time in the non-activated rotational thromboelastometry assay (NATEM) mode was &amp;gt;3600 s (minimum 1363 s) for AHA, 501 s (IQR: 430-632 s) for LA1, 533 s (IQR: 443-641 s) for LA2, and 500 s (IQR: 429-645 s) for the normal group. TGA showed that peak thrombin concentration was 16 nM (IQR: 13-29 nM) in the AHA group, 242 nM (IQR: 175-273 nM) in the LA1 group, 174 nM (IQR: 129-259 nM) in the LA2 group, and 190 nM (IQR: 125-288 nM) in the normal group. In the CWA, Ad|min1| was 2.1 (IQR: 1.7-3.2) in the AHA group, 8.7 (IQR: 8.0-9.5) in the LA1 group, 6.7 (IQR: 5.8-8.4) in the LA2 group, and 10.3 (IQR: 9.8-10.6) in the normal group. APTT-Mg/APTT was 1.23 (IQR: 1.18-1.29), 1.26 (IQR: 1.23-1.38), and 1.05 (IQR: 0.95-1.20) in the AHA, LA1, and LA2 groups, respectively. ROTEM, TGA, and CWA effectively differentiated between the LA1/2 and AHA groups. Clotting time in non-activated rotational thromboelastometry assay mode (ROTEM) and peak height (TGA) were better at distinguishing high-titer LA cases from LA-positive AHA cases. APTT-Mg/APTT could be suitable for distinguishing AHA from high-titer LA; however, it may not be useful for differentiating AHA from low-titer or suspected LA.

Interaction of Red Cell Indices and Blood Group in Diabetic Patients above 50 Years of Age

Introduction: A group of metabolic diseases with a hyperglycemic phenotype known as diabetes mellitus impact both industrialized and developing nations. It is brought on by deviations in insulin synthesis, action, or both.&#x0D; Research Objective: This study seeks to associate the interaction of red cell indices and blood group in diabetic patients above 50 years of age. This is a case control study in which a convenient sample size of 100 individuals were recruited from a consenting population at a private hospital in Port Harcourt, Rivers State, 50 of whom were diabetic, and the remaining 50 were not.&#x0D; Methods: Using the automated 3-part differential hematology analyzer Sysmex KX-21N, a Full blood count (FBC) and differential was obtained using (Sysmex Corporation Kobe, Japan) Standardization, instrument calibration, and analysis of sample was achieved in line with accordance with the manufacturer's manual to determine the red cell indices and grouping of cells using the tile approach to determine the ABO Blood Groups. Blood samples were taken from the veins using a vacutainer tube and needle according to Chesbrough's instructions, and then put in EDTA anticoagulated bottles with clear labelling. These samples were transported right away using the triple packing procedure to the lab and stored until testing at 4°C. Inferential statistics utilizing the students' statistical t-test were employed to compare the results.&#x0D; Results: Blood group O, with a total of 25 (50%), had the highest prevalence of diabetes mellitus, followed by blood group A with a total of 12 subjects (24%) and blood group B with a total of 8 subjects (16%) and blood group AB came in last with a total of 5 participants (10%), respectively. In this study, blood group O predominated over blood groups A and B (47% against 25% versus 17%, respectively). The least common blood type in this study was AB (11%). When the Packed Cell Volume, Haemoglobin, Red Blood Cell, Mean Cell volume, Mean Cell Haemoglobin, Mean Cell Haemoglobin Concentration, and Red cell Distribution Width were examined between diabetic and non-diabetic participants, no statistically significant (p&lt;0.05) difference was found.&#x0D; Conclusion: The findings of this study suggest that blood types O and AB are more prevalent in diabetics. We can deduce that a person's blood type may be a risk factor for developing this condition and may be useful in diagnosing it.

A-140 Evaluating the Performance of Various Lupus Anticoagulants Tests Among Different Coagulation Platforms

Abstract Background Lupus Anticoagulants (LAs), one of the antiphospholipid antibodies, are risk factors for arterial or venous thrombosis with complications such as stroke, acquired thrombophilia, and pregnancy loss. Detection of LAs is challenging for laboratories as the heterogeneity of antibodies and epitopes on protein in the phospholipids. Thus, no single test is sensitive for all LAs, and the International Society on Thrombosis and Haemostasis Scientific and Standardization Committee (ISTH) recommended laboratory should provide two test systems based on different principles, which dilute Russell's viper venom time (dRVVT) should be as the major test, and partnered with LA-sensitive activated partial thromboplastin time (aPTT). There are many different mechanisms of tests proposed by various manufacturers, however, the comparison of different platforms by different manufacturers on LAs remains unknown. In this study, we aimed to evaluate the performance of various LA tests among different platforms. Methods A total of 76 specimens from September to December 2022 were collected. dRVVT (dRVVTIL) and silica clotting time (SCTIL) were performed by ACL TOP 500 analyzer (Instrumentation Laboratory, US). In addition, ellagic acid-based aPTT (Siemens Dade Actin FSL reagent, aPTTAFSL), silica-based aPTT (Siemens Pathromtin SL reagent, aPTTPSL), and dRVVT (dRVVTsysmex) were performed by Sysmex CN-6000 analyzer (Sysmex Corporation, Japan). Analytical performances, agreements, and McNemar's test were calculated. Results All 76 specimens were originally tested by dRVVTIL and SCTIL, and showed that 30 specimens were positive for both assays, 6 were only positive for dRVVT, 5 were only positive for SCT, and 35 specimens were negative for both assays. In addition, all samples were further tested by dRVVTsysmex. Compared with the integrated use of dRVVTIL and SCTIL, dRVVTsysmex showed 100.0% (41/41) positive agreement and 91.4% (32/35) negative agreement. 3 specimens could only be tested positive on the dRVVTsysmex platform, of which one patient was diagnosed with systemic lupus erythematosus (SLE), another patient was diagnosed with infertility, and the other patient was diagnosed with antiphospholipid syndrome (APS). To further evaluate the performance of aPTT on LA screening, 44 specimens with either dRVVTIL, SCTIL and dRVVTsysmex positive were considered LA-positive and further tested by different aPTT assays. The positive agreement was 65.9% (29/44; 95% CI: 50.1–79.5%) and 77.3% (34/44; 95% CI: 62.2–88.5%) on aPTTAFSL and aPTTPSL, respectively, however, there was no significant difference between the positive agreement of two assays (P = 0.13). Conclusion Compared with the integrated use of dRVVTIL and SCTIL, dRVVTsysmex could achieve the same detection rate on LA. Furthermore, aPTTAFSL and aPTTPSL showed no significant difference between the positive agreements for LA-positive specimens.

Reticulated Platelet Count as a Diagnostic Tool in Immune Thrombocytopenia (ITP).

This study aimed to compare the reticulated platelet count between patients having thrombocytopenia secondary to autoimmune destruction (immune thrombocytopenia {ITP}), bone marrow failure, and healthy controls who presented to a tertiary care hospital in Karachi, Pakistan. A cross-sectional study wasconducted from February 2021 to October 2022 in the Department of Hematology, National Institute of Blood Disease (NIBD) Hospital in Karachi, Pakistan, that involved examining three groups: 30 patients with immune thrombocytopenia, 30 patients with thrombocytopenia secondary to reduced production from bone marrow, and 30 healthy controls. The study utilized the Sysmex XN-1000 (Hyogo, Japan: Sysmex Corporation) automated hematology analyzer to perform a complete blood count (CBC) test. Additionally, peripheral blood was stained with Leishman stain and examined under a microscope to eliminate pseudo thrombocytopenia and identify any abnormal cells or dysplasia. The immature platelet fraction (IPF) was then performed on Sysmex XN 1000 after ensuring adequate quality control. Finally, the data were analyzed using DATAtab (Graz, Austria: DATAtab) and SPSS version 25 (Armonk, NY: IBM Corp.). Of the ninety participants, the median age was 33 years with a range of 18-71 years. Patients with ITP had a significantly higher median IPF% (median=26.65, IQR=15-39.4) than thrombocytopenia due to bone marrow failure (median=9.25, IQR=4.55-14.30) and healthy controls (median=7, IQR=4.40-9.90), with a p-value of 0.001. The immune thrombocytopenia group demonstrated an increase in IPF% as platelet counts increased, indicating a significant moderate correlation between IPF% and platelets in these patients (r=0.438, p=0.016) and confirming that IPF% was an independent predictor for the detection of ITP. Reticulated platelet count may be a useful diagnostic tool to differentiate between ITP and thrombocytopenia caused by bone marrow failure. Because of its non-invasive nature, IPF is a valuable tool for expediting the management of thrombocytopenia associated with increased IPF.

Abstract 1026: A novel detection system of circulating tumor cells based on size-dependent enrichment with immunofluorescent staining of CK8/18 in advanced non-small cell lung cancer

Abstract Background: Detection of circulating tumor cells (CTCs) in the blood is a potentially noninvasive tool for cancer diagnosis, monitoring and prognosis evaluation. However, the CTC detection is difficult because of the rarity of CTCs, especially in non-small cell lung cancer (NSCLC), and the detection sensitivity of CTCs was reported to be less than 50% in NSCLC patients. Herein, we developed a novel CTC detection system and evaluated its performance in advanced NSCLC patients. Methods: Whole blood samples were collected from stage IV or recurrence NSCLC patients. CTC detection was performed with a novel system (Sysmex Corporation, Hyogo, Japan) by using size-dependent enrichment for capturing CTCs and immunofluorescent staining reagents including anti-cytokeratin (CK, AE1/AE3 or CK8/18) (specific for epithelial cells), anti-CD45 (specific for leukocyte) and Hoechst (for nucleus). Fluorescence intensity and cell images of each marker were obtained by Molecular Imaging Flow Cytometry (MI-FCM). Circulating cells that were positive for Hoechst and anti-CK but negative for anti-CD45 were defined as CTCs. To confirm whether CTCs detected were derived from lung cancer origin, CTCs obtained from patients with EGFR mutations were subjected to single cell sorting, whole genome amplification and PCR analysis for the mutation detection. Results: A total of 20 NSCLC patients (10 EGFR mutant, 10 EGFR wild-type) were enrolled in this study. Of these, CTCs were detected in 16 (80%) patients (number of CTCs, 1 to 66 in 4.5ml whole blood) when CK8/18 was used as epithelial cell staining, but in only 11 (55%) patients (1 to 6 CTCs) when using AE1/AE3. The detection sensitivity of CTCs based on CK8/18 was significantly higher compared to that based on AE1/AE3 in our novel system (80% vs. 50%, p = 0.03). The number of CTCs was significantly lower in EGFR mutation-positive patients than in EGFR wild-type patients (median number of CTCs, 3 vs. 22, p = 0.01). Furthermore, among 10 EGFR mutation-positive patients, EGFR mutations were detected in CTCs derived from 6 (60%) patients. All the mutation types detected in CTCs were matched with those identified in the corresponding primary tumors. Conclusion: Our novel CTC detection system demonstrated that the detection rate of CTCs (80%) was higher than those in previous studies. Further studies are needed for the clinical application of this system. Citation Format: Tetsuya Sakai, Yoshitaka Zenke, Mami Onishi, Eri Morita, Masatoshi Yanagida, Yuji Shibata, Hiroki Izumi, Kaname Nosaki, Hibiki Udagawa, Shingo Matsumoto, Kiyotaka Yoh, Koichi Goto. A novel detection system of circulating tumor cells based on size-dependent enrichment with immunofluorescent staining of CK8/18 in advanced non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1026.

ACCURACY OF AUTOMATED CELL ENUMERATION METHOD FOR VARYING CONCENTRATION OF WBCS FOR VARIOUS BODY FLUID SAMPLES

Background: Body fluids (BF) including peritoneal, pericardial, pleural, synovial and cerebrospinal fluids are now being analyzed by fully automated methods that are replacing the manual methods. The aim of this study is to assess the accuracy of automated instrument at varying concentrations of WBCs of various body fluid samples. Material and Methods: This cross-sectional study was conducted at the section of Hematology, Department of Pathology and Laboratory Medicine, the Aga Khan University, Karachi, Pakistan from November 2020 to April 2021. Forty body fluid samples with suspicion of infection including peritoneal, pericardial, pleural, synovial and cerebrospinal fluids (CSF) were analyzed to verify accuracy of white blood cells counts on fully automated XN-1000TM hematology analyzer (Sysmex Corporation, Kobe, Japan) against Neubauer chamber method (a manual method of differentiating WBCs via microscopy). The culture results of the body fluid samples were also analyzed. EP Evaluator version 10.3.0.556 (Data Innovations, LLC, VT, US) was used for statistical analysis with other supporting statistics such as Correlation Coefficient (R), Bias, Mean and Standard Deviation were included. Results: Eleven (n=11) of forty (n=40) body fluid samples showed high WBC count above the normal range. Samples with normal WBC (n=29) were also included in accuracy study to assess instrument performance at varying concentration of analyte. An average Error Index of -0.27 (-0.99 to 0.74) for WBC was obtained. Microbiological cultures grew Escherichia. Coli in 1 and Acinetobacter species in 1 CSF samples and Staphylococcus Aureus in 1 pleural sample. Conclusion: The study verified accuracy of varying concentration of WBC counting by fully automated Sysmex XN-1000 analyzer for various body fluid samples. Keywords: Body fluids, Validation, Automated analyzer, Performance specifications, Accuracy verifications, High white blood count, Infectious.

Effect of low-volume blood withdrawal on hematological biomarkers

Introduction&#x0D; The hematological module of the Athlete Biological Passport (ABP) consists of individual and longitudinal monitoring of hematological parameters biomarkers responsive to erythropoiesis alteration. Based on an adaptive Bayesian model, the ABP seeks to identify non-physiological variations that may correspond to different types of blood doping. In this context, the impact of a standard blood donation (500 ml) was evident on the ABP parameters, hence supporting the robustness of the model. However, there is only scarce evidence of single or multiple withdrawals of smaller volumes of blood, closer to a modern blood doping strategy. This study investigated the impact of low-volume blood withdrawal on hematological variables for anti-doping purposes.&#x0D; Methods&#x0D; Twelve healthy subjects (7 women and 5 men; 26 ±6 yrs, 170 ±7 cm; 65 ±7 kg) were recruited. After baseline measurement, 140 ml whole blood was withdrawn with subsequent measures of hematological parameters by flow cytometry (Sysmex XN-1000, Sysmex Corporation, Japan) for 3 weeks. Blood volumes (BV) and total hemoglobin mass (Hbmass) were determined by means of a validated CO-rebreathing method. Ferritin (FERR) concentration was additionally measured by chemifluminescence immunoassay technique. Individual profiles were generated using the official ABP adaptive model using the official software platform of the World Anti-Doping Agency (WADA). Mixed model repeated measures analyses were run for each parameter to determine whether changes in the dependent (hematological variables) variables differed over time (fixed factor).&#x0D; Results&#x0D; 43.2 ±8 g of hemoglobin was removed during the 140 ml blood withdrawal. A significant decrease in Hbmass (726 ±185 vs. 709 ±189 g; p = 0.007) and Red Blood Cell Volume (RBCV; 2190 ±521 vs. 2129 ±552 L; p = 0.028) was observed at D + 7 with a return to basal value at D + 14. No significant variations were observed for the primary and secondary variables of the ABP. No Atypical Passport Finding (ATPF) were generated in the individual ABP profiles. Conversely, FERR was significantly downregulated at D + 7 (95.8 ±76 vs. 87.1 ±76 μg/l; p = 0.018).&#x0D; Discussion/Conclusion&#x0D; While the ABP has demonstrated substantial advantages for traditional doping scenarios, our results challenge the indirect detection of low-volume blood withdrawals in a contemporary blood doping scenario. The impact of low-volume autologous blood transfusion on time trial performance has recently been demonstrated, supporting the need for novel biomarkers with extended detection windows. Hbmass could hence represent an additional marker to screen blood doping while technical issues for the use of CO-rebreathing remain. FERR as a marker of altered iron metabolism was there against shown to be sensitive and possibly suitable for outlining low-volume blood transfusion. The large inter-subject variability underlines the importance of individual longitudinal monitoring to provide sufficient sensitivity in the interpretation of hematological variations. Our study finally confirms that even low-volume blood withdrawals are associated with significant hematological changes while frequent blood samples and additional biomarkers are certainly required to better tackle blood doping strategies using such low-volume transfusions.

An Overview of Complete Blood Count Sample Rejection Rates in a Clinical Hematology Laboratory Due to Various Preanalytical Errors.

Introduction The Chughtai Laboratory collects blood samples for complete blood counts from various hospitals, emergency departments, ICUs, and through home sampling servicesall across the country. The preanalytical phase isan integral componentof laboratory medicine. A laboratory report has a key role in patient treatment and the clinician's decision in the management of the disease. Preanalytical errors are most frequently caused by the absence of a sample and/or inappropriate understanding of a test request, mislabeling, contamination from the sampling site, hemolyzed, clotted, insufficient samples, storage issues, and inappropriate blood to anticoagulant proportion or inappropriate choice of anticoagulant. Objective To identify the cause ofrejection rates ofthe complete blood countsamples and reduce the rejection rates by improving the accuracy of the results and loweringpre-analytical errors. Methods This cross-sectional study was done in the Hematology Department of Chughtai Laboratory's head office in Lahore between 19-06-2021 and 19-10-2021. Simple random sampling was applied to collect the data. About 3 ml of each blood sample was received in an ethylenediaminetetraacetic acid (EDTA) vial, inspected visually, run on Sysmex XN-9000 (Sysmex Corporation,Kobe, Hyogo, Japan), and was reviewed on peripheral smears. Results Out of 231,008 blood samples, 11,897 (5.15%) samples were rejected. The most common pre-analytical mistake was storage issues due to transportation delay (19.45%) followed by wrong medical records (19.16%), diluted samples (16.35%), incorrect tubes (16.01%), hemolyzed samples (15.13%), unlabeled samples (10.01%), and clotted sample (3.88%). Conclusion In the hematology department, the total rejection rate observed during the study period was 5.15%. Recognition of preanalytical errors and avoiding them will help uslower the sample rejection rateand raise the overall quality of laboratory management.

Frequency of Nucleated Red Blood Cells in the Peripheral Blood of ICU-Admitted Patients.

To evaluate the clinical significance of NRBCs in the peripheral blood of critically ill patients admitted to the ICU to find a cut-off to predict mortality. A cross-sectional study was carried out over a period of six months September 1, 2020, to March 31, 2021, in Lahore, Pakistan. A totalof 800 critically ill patients of both sexes in the age group of 18-70 years were included. Patients younger than 18 years and patients who underwentsurgery were excluded. A quantity of 3 ml of whole blood sample in an ethylenediamine tetraacetic acid (EDTA) vial from each patient was run on SYSMEX XN-9000 (Sysmex Corporation, Kobe, Hyogo, Japan) and the results were reviewed on peripheral smears. The incidence of NRBCs in ICU-admitted patients was 62.5% (500/800). The total number of NRBC-positive patients recovering after the treatment was 364 (72.8%). The overall mortality of NRBC-positive patients was 30% (150/500). It was significantly higher (p<0.001) than that of NRBC-negative patients (14%; 44/300).During treatment, the highest mortality rate was seen in patients due to malignancy (100%), followed by sepsis (58.8%). It was observed that the disease pattern and number of NRBCs were significantly different (p<0.001) among all disease groups. However, there was no statistically significant difference in NRBCs on the basis of gender (p >0.05). In our study, a cutoff of NRBCs of 2.50 showed a high risk of mortality with a sensitivity of 91%. The presence of NRBCs may predict mortality in critically ill ICU-admitted patients. Their presence in the blood may be regarded as a marker of severity suggesting a high risk of ICU death.

АНАЛИЗ ВОЗМОЖНОСТЕЙ ПРИМЕНЕНИЯ НОВОГО МЕТОДА ОПРЕДЕЛЕНИЯ УРОВНЯ D-ДИМЕРА В ПЛАЗМЕ КРОВИ У БОЛЬНЫХ С COVID-19 НА ГОСПИТАЛЬНОМ ЭТАПЕ

Introduction. The outbreak of the novel viral infection SARS-CoV-2 and its associated pandemic has generated significant interest among professionals regarding the role of D-dimer as a marker. D-dimer not only reflects the intensity of hemostatic and fibrinolytic reactions but also holds prognostic value. However, the standardization of D-dimer determination remains elusive, as the findings are known to depend on the completeness of fibrinolysis in the vasculature. Objective. This study aims to investigate the objective values of D-dimer levels, taking into consideration the concentration of its terminal fragments in plasma, specifically in patients with intravascular thrombosis associated with coronavirus infection. Materials and Methods. The study involved the examination of sodium citrate-stabilized plasma from 90 patients with severe COVID-19 who required respiratory support upon admission to the intensive care unit (ICU). The ICU mortality rate was 50% within 14 days. Additionally, a control group of 78 healthy individuals was included. The quantitative determination of D-dimer in human plasma was conducted using the Auto Red D-Dimer 700 (Helena Bioscience) and Sysmex CA1500 coagulation analyzer (Sysmex Corporation). Fibrinolysis was stimulated in vitro by streptokinase during sample preparation. The assessment was based on the analysis of blood plasma before and after the administration of the fibrinolysis activator. Results. Using the classical method of determination, D-dimer levels exceeded the cut-off value (300 ng/ml) in 61 out of 90 patients (67.8%). After fibrinolysis stimulation with streptokinase, this number increased to 66 (73.3%). Compared to the survival group, the fatal outcome group exhibited a 143.6% higher D-dimer level using the classical approach (p=0.2355). With the new approach involving streptokinase, the increase was 181.2% (p=0.0257). In one-fifth of fatal cases, the streptokinase-induced D-dimer concentration was more than twice the baseline obtained through the classical approach. Conclusion. The proposed approach for quantitatively determining D-dimer, which incorporates the use of streptokinase, provides more accurate data regarding the plasma concentration of D-dimer neoantigens. These findings establish a potential direction for standardizing such studies in clinical practice, particularly for determining treatment strategies for patients with COVID-19.