Clinical Pathology Laboratory Research Articles

Complete Blood Count and Biochemistry Reference Intervals for Healthy Adult Donkeys in the United States

Previous hematologic and serum biochemistry reference interval (RI) values have been established for donkeys in various geographic regions, life-stages, or for specific donkey breeds. The last extensive investigation establishing RIs for adult donkeys in the United States (U.S.) was published over three decades ago. We aimed to establish updated robust RIs using a reference population of apparently healthy adult donkeys from across the U.S. Standard sized (n = 102), miniature (n = 17), and mammoth (n = 1) donkeys from four different states were enrolled, with 20% of the study population including donkeys captured directly from the wild in Death Valley National Park, CA. RIs were established in accordance with the American Society for Veterinary Clinical Pathology and Clinical Laboratory Standards Institute guidelines. The findings will assist practitioners with the interpretation of their complete blood count and biochemistry panel results in U.S. donkeys. This study also highlights a comparison of results for some important analytes in U.S. donkeys compared to U.S. horses and previously established donkey RIs.

Perbedaan Derajat Aglutinasi Pemeriksaan Golongan Darah Metode Tabung Berdasarkan Konsentrasi Suspensi Sel 5% Segera Periksan Dengan Lama Penyimpanan 5 Hari

Background: ABO b;ood group examination is an examination carried out to determinate the type of blood group in humans that detects the presence of antigens on the surface of the red blood cell membrane by means of the Tube Method various consentrations of erythrocyte cell suspension,namely 5%, 10% and 15%. Research purpose: To determinate the effect of various consentrations of erythrocyte cell suspension with ready-to-use 0,9% NaCl and 0,9% table salt solvent on the degree of agglutination in blood group examination Tube method. Research method: This research is a pure experimental study with a total sample 41 people, this study was conducted on January 16-20, 2023 at the Muhammadiyah Palembang IKesT Clinical Pathology Laboratory. Results: The results showed that the blood group examination was done by the tube method with a concentration of erythrocyte cell suspension, namely 5%, 10% and 15% with 0,9% NaCl solvent ready-to-use and 0,9% NaCl, table salt solvent resulting in a degree of agglutination of +4. The Kruskal-wallis test result in the Cell Grouping method obtained 0,000 and in the Serum Grouping method obtained 0,051. Conclusion: The conclusion in this study is that there is no effect of various consentrations of erythrocyte cell suspension with ready-to-use 0,9% NaCl and 0,9% NaCl Solvent Kitchen Salt on the degree of agglutination in blood group examination Tube Method so that 0,9% NaCl solvent from table salt can be used as an alternative to ready-to-use 0,9% NaCl reagent in blood group tests by tube method. 
 Keywords: Blood type, Aglutination

Risk management in a clinical pathology laboratory: Application of brainstorming and probability analysis for risk reduction

ABSTRACT Introduction: Risk management includes identifying various risks, assessing the probability of occurrence, and evaluating the severity of their consequences. As clinical laboratories are integrally involved in patient care, risks in the laboratories could present grave consequences in some instances. This study aimed to utilize simple techniques for risk management in a clinical laboratory. Materials and Methods: All potential risks in the pathology laboratory of a tertiary-level hospital were identified and classified into natural calamity, environmental, manpower-related, pre-analytical, analytical, post-analytical, and laboratory hazard-related risks through a brainstorming session. The probability of occurrence of each risk was estimated from departmental and hospital records. The possible impact of risk (score 1–10) was categorized into catastrophic, critical, serious, minor negligible, and insignificant. The unweighted risk score was calculated by multiplying the probability of occurrence and impact score. Results: Inadequate sample-to-anticoagulant ratio had the highest probability of occurrence (22.85%), followed by quantity insufficient for analysis (7.30%) and laboratory information system (LIS) breakdown (6.58%). The highest unweighted risk score in our study was inadequate sample-to-anticoagulant ratio (score 91.40), followed by improperly labeled samples (score 35.61), manpower competency issues (score 32.88), sample insufficient for analysis (score 29.20), and LIS breakdown (score 26.30). Conclusion: We found that among all the categories, risks involving the pre-analytical phase had the highest risk scores. The other important risks included manpower competency issues requiring continued on-the-job training of staff as a risk reduction strategy. Brainstorming and probability analysis could be easily used for risk management in a clinical laboratory.

Serum Interleukin-6 Levels are Positively Correlated with the Severity of Acne Vulgaris

Acne vulgaris is a chronic inflammatory disease that affects the pilosebaceous unit. This disease occurs on the skin of the face, body, back, and other areas of the body, and can leave scars on the skin which can be temporary or permanent. Recent data shows that the inflammatory process is the initial process in the pathogenesis of acne, several pro-inflammatory cytokines, one of which is interleukin 6 (IL-6), which plays a role in acne vulgaris. Analytical observational study with a cross-sectional approach. The research was carried out from February - November 2023 at the Skin and Venereology Polyclinic, RSUP Prof. Dr. I.G.N.G. Ngoerah Denpasar and the Clinical Pathology Laboratory of Prof IGNG Ngoerah Hospital. The sample consisted of 47 samples of patients with acne and 20 samples without acne aged 8 - 45 years who were selected through consecutive sampling according to inclusion and exclusion criteria. A sample of 3 milliliters of venous blood is taken which will then be checked for IL-6 with an ELISA kit. Data analysis with SPSS version 24 and p value <0.05 is significant. The mean IL-6 serum levels in the acne group was 110.2 ± 88.3 pg/ml while in the non-acne group it was 26.7 ± 23.7 pg/ml. There was a significant difference in the category of IL-6 levels in the acne and non-acne groups (95% CI 55.1 - 155.3, p<0.001). The correlation analysis of IL-6 levels with the severity of acne was strongly positive (r=0.951, p<0.001).

Lesson Learned from Mass Antibody Rapid Diagnostic Used in the Early COVID-19 Pandemic in Indonesia Contributors.

Laboratory examination is extremely important in handling the COVID-19 pandemic. In the first era of the pandemic, the molecular and antigen tests were limited. Hence, at that time, it was necessary to carry out antibody Rapid Diagnostic Tests (RDT). However, many antibody RDTs were yet to obtain Food and Drug Authorization (FDA)'s approval. Therefore, The Indonesian Association of Clinical Pathology and Medical Laboratory (PDS PatKLIn) decided to conduct a validity test of RDT antibodies to find out the quality of SARS-CoV-2 diagnosis performance based on these RDTs used. This is a descriptive observational design with diagnostic analysis. The retrospective secondary data were collected from 34 provinces in Indonesia from May to June 2020. Data analysis was carried out on the sensitivity and specificity values of each antibody RDT brand to the RT-PCR result and analyzed descriptive data. The amount of secondary data of antibody RDT and RT-PCR results collected was 139,908, consisting of 59 RDT brands of which 44% were authorized by The Indonesian COVID-19 Response Acceleration Task Force (Gugus Tugas Percepatan Penanganan COVID-19 Indonesia). There were huge variations of SARS-CoV-2 antibody RDT performance between total antibody types (sensitivity 59.18%, specificity 62%), IgM RDT (sensitivity 16-100%, specificity 7-97%), and RDT IgG (sensitivity 33-96%, specificity 19-100%). The variations in the RDT antibodies'performance can cause errors in diagnosis leading to significant material and immaterial losses. Therefore, cooperation from various parties is needed for the pre- and post-marketing surveillance process to assess the performance and the characteristics of each RDT kit and other diagnostic methods to assist the rapid pandemic response process.

How clinical laboratories select and use Analytical Performance Specifications (APS) in Italy.

Even if the topic of the analytical quality required to provide laboratory results "fit for purpose" exists since the beginning of the modern medical laboratory, there is the suspect that the expression "Analytical Performance Specifications" (APS) is not well-known. To investigate this aspect a survey was conducted. A questionnaire with seven questions related to the knowledge about the topic, the sources of information and the criteria used by the laboratories to set the APS and their applications was prepared. It was distributed to all the clinical pathology laboratories of Lombardy Region (143) and to the members of SIBioC Laboratory Medicine (excluding Lombardy). We received 201 replies: 127 from Lombardy and 74 from the rest of Italy. Fifteen percent declared to ignore the meaning ofAPS and only 64 % of those knowing the meaning of theterm declared to use them in the daily practice. The state-of-the-art was the principle used more frequently to setAPS(about 48 %) followed by biological variation (41 %), and APS were typically applied to set goals for Internal Quality Control for selected measurands. Usually imprecision or total error APS were used, much less frequently uncertainty APS. In fact only 27 % of the laboratories declared to have calculated the measurement uncertainty for part or the majority of their measurands. Even considering the limits of a survey that relies upon self-declarations, it appears clearly that, at list in Italy, there is some work to be done to promote the concept and the use of APS.

Measuring the Level of Agreement for Lactate Measurements in Hispaniolan Amazon Parrots (Amazona ventralis) Among 2 Point-of-Care Analyzers and a Benchtop Analyzer.

Lactate is an important biochemistry analyte used in human and veterinary medicine to assess tissue perfusion and can be used as a prognostic indicator for certain disease conditions. Whereas lactate is commonly measured using "patient-side" handheld meters, these meters have not been validated for companion avian species. The purpose of this study was to measure the level of agreement between 2 commercially available point-of-care lactate meters and a laboratory benchtop blood analyzer in Hispaniolan Amazon parrots (Amazona ventralis). Blood samples were collected from 20 adult parrots at Louisiana State University by drawing 1.5 mL of blood from the right jugular vein. One drop of whole blood was used for the Lactate Plus analyzer and the remainder of the sample transferred into a lithium heparin microtainer. From the blood in the microtainer, 0.2 mL whole blood was analyzed using the epoc Blood Analysis System, and the remaining sample was centrifuged to obtain plasma that was immediately frozen at -80°C (-112°F) and submitted to the Texas A&M University Clinical Pathology Laboratory for analysis on the VITROS 4500 benchtop analyzer. Bland-Altman agreement plots and Passing-Bablok regression were used to measure the level of agreement between the methods. There was poor agreement between all 3 methods with mean percentage differences in lactate concentrations ≥22% (epoc and Lactate Plus: 33.6% [95% CI: 27-40]; epoc and VITROS 4500: 55% [95% CI:52-58]; VITROS 4500 and Lactate Plus: 22% [95% CI:16-28]). Based on these results, the point-of-care meters tested in this study are not interchangeable, and separate reference intervals were calculated for each method. Blood lactate concentrations may have more utility in tracing lactate trends over time in an individual rather than being able to utilize this information at 1 time point for disease diagnosis and prognosis.

Creating Efficient Workflows for Electron Microscopy Laboratories with Automated Specimen Preparation

Abstract Electron microscopy (EM) is essential to the biological and biomedical sciences and clinical diagnostic pathology. Electron microscopy specimen preparation is laborious and time-consuming, with transmission EM (TEM) chemical preparation typically requiring 1–3 days, while volume electron microscopy (vEM) takes 3–5 days of tedious manual reagent exchanges every few minutes or hours. This places a considerable burden on laboratory scientists with the ongoing demand for TEM, and the rapidly growing demand for vEM due to its potential to revolutionize structural biology, connectomics, and related fields. This burden is exacerbated by a shortage of trained electron microscopy scientists as current staff retire, and few enter the workforce. This report provides four case studies to illustrate how automated and faster specimen preparation workflows using mPrep™ Automated Specimen Processors (ASP-1000™ and ASP-2000™, Microscopy Innovations, LLC) free electron microscopy staff in academic and pre-clinical research labs and a clinical pathology laboratory.

Performance Evaluation of the Commonly-Used Portable Cholesterol Sensors for Telehealth Services in the Unreached Communities.

Portable medical sensors play an important role in healthcare services, especially in rural communities. Many telehealth systems use these devices for providing patients' vital information from a distance to remote doctors. Erroneous data will not only mislead the remote doctor for correct diagnosis but it will cause health threats to these unreached community people. Therefore, it is very important to identify good sensors with an acceptable level of accuracy but within the affordable price of the available sensors in the market. This study aims to identify quality portable cholesterol sensors with high accuracy with the reference of the Japanese clinical pathology laboratory as a gold standard. We have considered cholesterol sensors that measure total cholesterol for this study that are commonly used in the developing countries of Asia. We found that out of four, three of them were very much erroneous and cannot be recommended even for primary healthcare.

An evaluation of the commonly used portable medical sensors performance in comparison to clinical test results for telehealth systems

Background and ObjectiveOne of the primary challenges faced by telehealth systems is the accurate transmission of patient information to remote doctors. In this context, portable medical sensors deployed at the remote patients' end play a crucial role in measuring vital information. There are many sensors available in the market. However, the accuracy of the sensors has been always a concern. The objective of this study is to verify different sensors and create awareness for using accurate sensors to avoid misdiagnosis for the patients’ safety. MethodsThis study considered the test result of a Japanese clinical pathology laboratory as the reference gold standard. The clinical pathology laboratory uses 1) Hexokinase UV method for blood glucose, 2) Enzymatic Determination method for cholesterol, 3) Automatic Analyzer (EDTA-2 K) of Hemoglobin, and 4) Uricase POD method for uric acid. To assess the performance of a medical sensor, its test results were compared to the gold standard test results obtained from the laboratory using the same sample. A Normalized Root Mean Square Error (NRMSE) threshold of less than 0.2 was established as the criterion for determining whether the medical sensor's performance fell within an acceptable range. ResultsAmong the eight most commonly used blood glucose devices in the Asian market, only one device was deemed acceptable with NRMSE less than 0.2. However, all four devices found in the Japanese market showed their acceptability. In the case of cholesterol, hemoglobin, and uric acid devices, only a limited number of items were available in Asian markets. Some of the hemoglobin and uric acid devices were found to be somewhat acceptable, while all the cholesterol sensors were found erroneous. ConclusionsThis study has clearly shown the issues with the portable medical sensors and recommends the device approval authority of each country to approve sales of the quality sensors only for patients’ safety.

OVERVIEW OF BACTERIAL PATTERNS IN TABANAN GENERAL HOSPITAL

AbstractBackground: Resistant bacterial infections will affect the results of therapy, costs, spread of disease, and duration of illness. To minimize the losses incurred, it is necessary to monitor resistant bacterial and the use of antibiotics. Objective: To be able to identify, monitor, map and overcome antibiotic resistance and restrict bacterial transmission in hospitals, especially at the Tabanan General Hospital. Method: This research was conducted using a cross-sectional method using samples of inpatients and outpatients sent by clinicians to the Clinical Pathology Laboratory, Microbiology Sub Lab in period of January to December 2022. Results: Of the 590 positive culture samples in 2022, the majority were caused by Escherichia coli (15.25%) and 62.22% were ESBL. Apart from that, 8.31% were found to be Staphylococcus aureus and 53.06% were MRSA. Conclusion: The percentage of Escherichia coli bacteria that produce the ESBL enzyme is still high (62.22%). The percentage of Staphylococcus aureus bacteria that produce the MRSA enzyme is still high (53.06%). The antibiotics Meropenem, Amikacin, and Tigecycline are the antibiotics with the highest sensitivity to ESBL. The antibiotics Nitrofurantoin, Linezolid, and Quinupristin/Dalfopristin are the antibiotics with the highest sensitivity to MRSA. Keywords: bacterial patterns, description of bacterial distribution, antibiotic resistance

Korelasi Kadar HbA1c Dengan Kadar Kreatinin Pada Pasien Diabetes Melitus Kronis

Uncontrolled diabetes mellitus will lead to various chronic complications, both microvascular and macrovascular complications. HbA1c examination is effective for long-term monitoring of blood glucose for people with diabetes mellitus. A high HbA1c value is used as a measure of the development of complications of diabetes mellitus. Diabetic nephropathy is the most common complication of diabetes mellitus which can end in kidney failure resulting in damage to the glomeruli. Indications to determine kidney function impairment is creatinine examination. This study aims to determine the correlation between HbA1c levels and creatinine levels in patients with chronic diabetes mellitus at the Clinical Pathology Laboratory of the Bajawa Regional General Hospital. Analytical observational research with an approach cros- sectional using techniques purposive sampling 35 chronic DM patients. Test results who squares shows that the value of p = (0.687) > α (0.05), thus it can be concluded that there is no relationship between HbA1c levels and Creatinine levels in patients with chronic diabetes mellitus.
 
 Keywords: HbA1c, Creatinine, Diabetes Mellitus.

Development of a targeted proteomic assay for the quantitation of neurofilament light in cerebrospinal fluid: Correlation with Simoa‐based quantitation across neurodegenerative diseases and centres

AbstractBackgroundCerebrospinal fluid (CSF) neurofilament light (NfL) is a dynamic biomarker of neurodegeneration that is used for supporting clinical diagnosis and monitoring disease‐modifying therapies. Development of new analytical tests has been limited, with quantitation of NfL in biomarker studies largely restricted to immunoassay methods. Targeted mass spectrometry is a highly specific technique that can be easily translated into existing proteomic and clinical pathology laboratories. In this multi‐centre study, we have developed two targeted mass spectrometry assays which were compared to Simoa, the gold standard method for NfL quantitation. The developed methods included immunocapture of intact NfL protein or specific peptides (immune‐peptide capture) prior to targeted proteomic analyses. We discuss the development, validation and merits of each technology to quantitate NfL in CSF from healthy controls and several neurodegenerative diseases.MethodCSF was collected from patients with a range of neurological diseases at the National Hospital for Neurology and Neurosurgery (London, UK). Clinical diagnosis was determined by their treating clinician according to consensus clinical criteria. NfL concentrations were measured by Simoa (Quanterix, USA) and two immunoprecipitation‐targeted mass spectrometry (IP‐MS) assays developed by a collaboration between UCL and Washington University in St Louis. To evaluate replication of results across centres and mass spectrometry approaches, results were compared using Spearman’s correlation analysis and Bland‐Altman testing.ResultsA high correlation was observed for CSF NfL levels between Simoa and both IP‐MS assays (immune‐peptide capture, r = 0.90, P<0.0001 and immune‐protein capture, r = 0.88, P<0.0001). Relative differences in mean NfL concentration between clinical groups remained the same across the analytical methods, with the highest and lowest NfL concentrations observed in the Huntington’s disease and the healthy control groups respectively. Interestingly, differences in the extent of correlation between Simoa and IP‐MS were observed between the different clinical groups (r = 0.37 ‐ 0.94).ConclusionImmunoprecipitation‐targeted mass spectrometry quantitation of NfL correlates highly with NfL Simoa measures across a range of neurodegenerative and neuroinflammatory diseases, and is replicable between laboratories using different immunoprecipitation‐targeted mass spectrometry approaches.

Myelomatous Ascitis, a Rare Presenting Symptom of Multiple Myeloma

Abstract Introduction/Objective In 2022, plasma cell neoplasms accounted for approximately 1.8% of all new cancers in the United States with an estimated 34,470 new cases of multiple myeloma. Diagnosis is often straight forward in ~80% of cases by performing serum protein electrophoresis (SPEP), serum immunofixation (SIFE), and/or urine immunofixation (UIFE) with M-spike quantification. The diagnostic challange is in the remainder of asymptomatic myeloma cases (~20%) which are found incidentally. Specialized laboratory technicians are often the first step in identifying plasma cells in body fluids and/or peripheral blood smears. These incidental findings on routine labs, paired with the keen eye of a specialist, are one of the first steps in early detection of multiple myeloma. Methods/Case Report A 62 year-old male, originally from Cambodia, presented to an outside hospital emergency department with abdominal distention and pain. His past medical history was significant for dialysis-dependent end- stage renal disease and insulin-dependent diabetes mellitus type 2. Abdominal CT showed ascites with no other diagnostic findings. Cardiac, renal and hepatic etiologies of ascites were ruled out. Body fluid analysis of the ascites in the Clinical Pathology laboratory revealed numerous plasma cells. Cytologic examination, after consultation with Clinical Pathology, also revealed numerous plasma cells. Subsequent SPEP and SIFE disclosed an IgA lambda monoclonal protein. A diagnosis of multiple myeloma was confirmed by bone marrow biopsy with FISH studies. Results showed several high-risk abnormalities including translocations of 4::14, 14::16, and 14::20, denoting a “triple hit” multiple myeloma. Results (if a Case Study enter NA) NA Conclusion Plasmacytosis in ascites is an extremely rare initial presentation for multiple myeloma. With only a few cases reported in the literature, many technicians and pathologists might easily overlook plasma cells on routine cytology, especially with normal imaging and no previous diagnosis of myeloma. In this case, flagged peritoneal fluid by a skilled medical laboratory technician was key to making a diagnosis. Without any signs of a plasmacytoma or lytic bone lesions on imaging, a normal CBC with non-specific labs, and an unsuspecting surgical pathology report, multiple myeloma was not on the differential. Fast interdepartmental consultation, communication, and teamwork between surgical pathology and clinical pathology, lead to a rapid diagnosis and prompt treatment of this patient.

Clot-Based Assay for Factor XI (FXI) Activity Is Highly Sensitive to Lupus Anticoagulant (LA) and Heparin with Potential Misdiagnosis of FXI Deficiency: The Importance of Recognizing Non-Parallelism

Introduction: Clot-based assays are employed to estimate coagulation factor activity in clinical pathology laboratories. These assays are PT- or aPTT-based and utilize dilutions of the patient's plasma in factor-deficient plasma. The curves of activity for each dilution follow a parallelism or non-parallelism pattern and the latter is typically seen in the presence of Lupus Anticoagulant (LA). Identification of this pattern is critical to avoid misdiagnosis of factor deficiency in patients who actually have an inhibitor. Materials and Methods: We observed low Factor XI (FXI) levels in patients with LA, while other factors were not affected. In order to determine the reason(s) why FXI levels are sensitive to LA, we collected 34 plasma samples from patients with dRVVT confirm ratio > 1.2, and estimated FVIII and FXI levels. FXI activity was also estimated with Dade ® Actin ® FS, a reagent that is not sensitive to the LA. The coefficient of variance (CV) for both FXI and FVIII levels were compared. To estimate the effects of heparin, plasma was spiked (0.1-1.0 IU/ML) and tested in the PTT, FXI, and FVIII assays. We tested with Dade ® Actin ® FS 4 known normal patients, 3 known FXI deficient patients, 20 known plasma positive for LA, and 10 with heparin. All assays were performed on a Siemens CS-5100 analyzer. Results: The results show that FXI is highly sensitive to LA, with non-parallelism and high CV, not observed with FVIII activity. As expected, FXI levels were not affected when LA-insensitive Dade ® Actin ® FS was used. Heparin (0.3-0.7 IU/ML) also showed non-parallelism in FXI activity, but not for FVIII. Only the highest dose of heparin (1.0 IU/ML) affected FVIII levels. Notably, the sensitivity of FXI activity to LA and heparin was explained by a few dilutions of the patient's plasma (5X) in the FXI assay, compared to other coagulation factors (20X). Of note, less diluted plasma is required for FXI activity because relatively higher levels of FXI is required to sustain and produce clot in the FXI-deficient plasma. Conclusion: This simple, yet not recognized explanation based on a literature search, is helpful to increase awareness of this analytical variable. It is also important to avoid misdiagnosis of FXI deficiency in patients with LA.

Patterns of germs before, during and after the COVID-19 pandemic in Intensive Care Unit (ICU) patients at Dr. Wahidin Sudirohusodo, Makassar, Indonesia

Background: The intensive care unit (ICU) is a unit that cares for patients with a high risk of infection, while viral infections are one of the predisposing factors for someone being easily infected with bacteria. This research aims to see a picture of germ patterns before, during and after the COVID-19 pandemic. Methods: This research is descriptive with a retrospective approach. The research sample used secondary data from microbial culture results of clinical specimens (blood, sputum, pus, urine, etc.) from ICU patients for the period 2019-2022 at the Clinical Pathology Laboratory of Dr. Wahidin Sudirohusodo Hospital, Makassar. Data is grouped into Gram-positive and Gram-negative bacteria, based on the highest order, and presented as tables and graphs. Results: There were 440 isolates, 80 Gram-positive isolates (18%) and 360 Gram-negative isolates (82%). The five most common types of isolates were dominated by the Gram-negative group in the last 4 years, respectively: Burkholderia cepacia 92 (21%), Acinetobacter baumannii 89 (20%), Klebsiella pneumoniae ss. pneumoniae 58 (13%), Pseudomonas aeruginosa 37 (8%), Escherichia coli 37 (8%), and Staphylococcus aureus ss. aureus was the 6th most common, with 20 (5%) isolates. The majority of isolates identified were obtained from blood, as many as 192 (44%), sputum 136 (31%), pus 52 (12%) and urine 8 (2%). Bacteremia was the largest cause of infection by Burkholderia cepacia, 91 (47%), while sputum was dominated by Acinetobacter baumannii 49 (26%). Conclusion: The germ pattern in the ICU before, during, and after Gram-negative bacteria dominated the COVID-19 pandemic. Bacteremia is the largest proportion of causes of infection.

ASSESSMENT OF ALL TYPES OF LABORATORY ERRORS IN THE TERTIARY CARE CARDIAC FACILITY LABORATORY

A prospective examination of laboratory errors was performed during three months from July to September 2019 at the Rawalpindi Institute of Cardiology (RIC), a 272-bed tertiary care hospital serving the Punjab region and parts of Azad Jammu and Kashmir (AJK) and Khyber Pakhtunkhwa (KPK). The study aimed to thoroughly evaluate error incidence rates across the complete testing cycle, considering the pre-analytical, analytical, and post-analytical phases. During this time, nurses, physicians, and medical assistants from various hospital wards submitted samples and request forms to the clinical pathology laboratory at RIC. For a total of 17,917 patients, 73,540 tests were carried out. The overall observed laboratory error rate for the entire testing procedure (TTP) was 1.43%. Notably, pre-analytical mistakes made up the most significant percentage (0.82%), followed by post-analytical errors (0.51%) and analytical errors (0.10%). The data were carefully examined, and the results were presented using figures, charts, and tables to show the prevalence and magnitude of various mistake types. According to the distribution pattern, overall error rates had significantly decreased over the previous ten years. Despite this development, the largest error prevalence was still seen in the pre- and post-analytical processes. Interestingly, errors were frequently found during the pre- and post-analytical procedures outside the lab. To reduce the likelihood of mistakes, RIC undertook several initiatives, and the study highlighted these efforts, highlighting the significance of the laboratory testing procedure for mistake prevention throughout all testing phases.In addition to offering insightful information about the frequency of errors at RIC, this three-month descriptive analysis highlighted ongoing efforts to improve patient safety and lower laboratory errors. The observed gains over the past ten years are evidence of the success of adopted tactics. The results show how important it is to maintain constant watchfulness and employ methodical procedures to guarantee the accuracy and dependability of laboratory testing procedures, improving patient care and safety.

Correlation between Serum Levels of Vaspin, BMI, and Waist Circumference on Obese and Non-Obese Subjects

Obesity is the accumulation of excessive fat resulting from an imbalance in energy intake compared to energy used for a long period. Vaspin is secreted in adipose tissue and may exhibit compensatory responses related to obesity, insulin resistance, and type 2 diabetes mellitus. This study aimed to determine vaspin levels in obese and non-obese subjects. An observational research design with a cross-sectional approach was used in this study. A total of 80 research subjects were included in this study. The research was conducted at the Clinical Pathology Laboratory at the Department of Clinical Pathology, Hasanuddin University Hospital in January 2022. Kolmogorov-Smirnov test was used to determine the normality of the data. Chi-Square, unpaired T-test, Mann-Whitney test, and Spearman test were used as statistical tests. This study showed a significant difference in anthropometric parameters between the obese and non-obese groups (p < 0.001). There was a significant difference in the mean levels of vaspin between the obese and non-obese groups (707.68±774.47 ng/mL vs. 510.60±460.08 ng/mL, p=0.004). There was a positive correlation between vaspin levels and waist circumference (r = 0.245, p=0.004) and between vaspin levels and BMI (r = 0.322, p=0.028). The serum levels of vaspin tended to be higher in obese subjects compared to non-obese subjects. The positive correlation between vaspin levels and BMI and waist circumference indicated that a higher BMI and/or waist circumference led to a higher vaspin level.

Polymorphonuclear-Mononuclear Ratio in Peripheral Blood as Hematologic Malignancy Predictor in Pancytopenia

Pancytopenia is a laboratory finding of decreased hematological cells characterized by hemoglobin of <13.5 g/dL for males or <11.5 g/dL for females, leukocytes of <4x109/L and platelets of <150x109/L. The data from Dr. Moewardi Hospital reported 56 cases of pancytopenia in 2020. Follow-up tests such as reticulocyte test, Bone Marrow Puncture (BMP), or bone marrow biopsy are needed to determine the cause of pancytopenia. This study aimed to assess the performance of the Pm/M ratio of peripheral blood as a screening instrument to predict the cause of pancytopenia. A cross-sectional study was carried out on pancytopenia patients undergoing laboratory tests at the Clinical Pathology Laboratories of Dr. Moewardi Hospital from January 2020 to June 2021. The cut-off point of the Pm/M ratio was determined by ROC and AUC curves. The results were presented in a 2x2 table. The Pm/M ratio <0.91 as a predictor of hematological malignancy had a sensitivity of 82.9%, specificity of 82.9%, PPV 82.9, NPV 82.9%, LR positive 4.833 and LR negative 0.207. The Pm/M ratio can be used as a screening biomarker to predict the cause of pancytopenia before performing BMP and to distinguish between hematological malignancy and non-hematological malignancy.

Prospective Molecular Characterization of Multiple Myeloma Patient Samples Identifies High-Risk Patients and Informs Treatment Sequences through Resistance Mechanisms to Immunotherapies

Introduction. Multiple myeloma (MM) genetic diagnostics has traditionally been performed using fluorescence in situ hybridization (FISH), giving prognostic information based on immunoglobulin (Ig) translocations and key copy number abnormalities such as 1p, 1q, and 17p. However, in recent years it has become clear that more mutations, biallelic events, and focal deletions are also prognostically important and can be captured better through molecular techniques such as DNA sequencing. In addition, as targeted and immunotherapies become standard, monitoring the molecular targets will be important in determining the optimal sequence of therapies. Methods. Bone marrow aspirates from patients with MGUS/amyloid (n=3), smoldering MM (n=3), newly diagnosed MM (n=15), or progression/relapse MM (n=33) were collected. All patients were enrolled in the Indiana Myeloma Registry and consented for use of data for research purposes. CD138+ MACS cell separation was performed in a clinical pathology laboratory and DNA was extracted from CD138+ cells and patient-matched buccal swabs. 100 ng DNA from both the tumor and control sample were processed using the HyperPlus kit (KAPA Biosystems) and hybridized to the Myeloma Genome Project (MGP) sequencing panel in a CLIA-certified laboratory. The panel consisted of 228 genes for mutation and copy number analysis, as well as the immunoglobulin and MYC loci for translocation analysis. Samples were sequenced using paired-end reads to a median depth of 1063x. Data were analyzed using a clinical workflow and genomic abnormalities reported by a pathologist. Results. Of the 54 patients, Ig translocations were detected in 30 and included the known t(4;14), t(6;14), t(11;14), t(14;16) and t(12;14) as well as rare Ig translocations to partners such as GADD45B, LYN, PPCDC, and RCBTB2, of which over-expression of GADD45B and LYN are potentially targetable. A complex t(1;8;11;14) was also detected involving CCND1, MYC and the IGH loci which would not have been detectable by FISH. High-risk markers, such as gain/amp1q (22.2%) and deletion/mutation TP53 (16.6%) were detected including biallelic TP53 alterations in 5 patients (9.3%), which were mostly deletion plus mutation (4/5) and would not have been detected by traditional techniques. Of the MM patients who had been previously treated (n=33), 27 had received an IMiD and 30 had received a proteasome inhibitor. One patient showed mutation of CRBN (p.N236D) in conjunction with del3p and another patient with mutation of CUL4B (p.E277Ter) with delX indicating complete inactivation of the genes as a resistance mechanism to IMiDs. In addition, 17 patients had received an immunotherapeutic regimen including anti-CD38 or anti-BCMA. Of those that had received an anti-CD38 treatment (n=17), three had deletion of 4p (the location of CD38) including one patient, who had received three previous anti-CD38 regimens, with focal deletion of CD38. Six patients had also received anti-BCMA therapy, of which one had a mutation in the gene encoding BCMA, TNFRSF17, resulting in premature termination within the extracellular domain (p.S48Ter) in 42% of cells. This sample came from a patient who relapsed after an initial response to a BCMA CAR-T, followed by a progression occurring quickly while on a BCMA bispecific antibody Teclistamab, suggesting the loss of BCMA expression as a mechanism of resistance in both treatments. Interestingly, two patients harbored somatic mutations in the transmembrane domains of either CD38 (p.S29I) or TNFRSF17 (p.L60S) prior to the start of the respective treatments indicating that not all mutations seen in these genes are a result of treatment resistance. Conclusions. DNA sequencing approaches are cost-effective alternatives to FISH, the current standard diagnostic technology, to detect genomic abnormalities in multiple myeloma and they can identify clinically meaningful markers related to prognosis and treatment, particularly in the emerging era of immunotherapeutics where antigen evasion is a key mechanism of molecular resistance.