Clotted Samples Research Articles

Appraisal of six sigma in pre-analytical phase of clinical biochemistry laboratory

Objectives: The aim of the study is to measure the performance of pre-analytical phase of a clinical biochemistry laboratory using sigma metrics and the six sigma scale. Materials and Methods: The study included documented data of blood sample rejection from March 2023 to February 2024 and follow-up data from March 2024 to August 2024. International Federation of Clinical Chemistry and Laboratory Medicine developed Quality Indicators (QIs) used are QI-9 Wrong tubes; QI-10 Hemolyzed samples; QI-11 Clotted samples; QI-12 Insufficient samples; QI-14 Damaged samples in transport; and QI-15 Mislabeled samples. Based on “Six Sigma Quality Design and Control” established by Dr. Westgard, the sigma metric was calculated for the above-mentioned QIs. Statistical analysis: Obtained data were entered and analyzed using Microsoft Excel 2021. Results: Out of 162,380 received samples, 547 samples were rejected as not satisfied with the sample acceptance criteria. The most common pre-analytical error in the observed QIs is hemolyzed samples (458), followed by insufficient sample volume (55). The Sigma score of QI-10 was determined to be 4.81, whereas QI-9, QI-11, QI-12, and QI-15 were well maintained and graded excellent. Following training sessions, the follow-up month revealed a sigma score of 4.98 for QI-10. Conclusions: Six sigma metrics are a competent means to measure the performance of pre-analytical QIs in a clinical biochemistry laboratory. The observed QIs were effectively managed (>4 σ).

An In Vitro Study of 355‐nm Laser Ablation of Atherosclerotic Lesions Model

ABSTRACTA study of 355 nm laser with high pulse energy across various types of atherosclerotic lesion models is presented. The 355 nm laser pulses (10 ns) are delivered via a single fiber (600 μm diameter), and the ablation of calcified tissue, lipid tissue, and thrombus‐like tissue are studied under varied laser fluence (40–70 mJ/mm2) and repetition rate (5–30 Hz). The contact and noncontact ablation processes of chicken tibia samples (calcified tissue) are compared at 60 mJ/mm2 and 30 Hz, and the size of ablation particles is in the range of 0.1–1 μm. At the same repetition rate, the advancement rate of tricalcium phosphate samples reaches 150 μm/s at 70 mJ/mm2. Calcified and lipid models demonstrate predictable increases in ablation with higher laser fluence and repetition rate. The fresh porcine blood clot samples exhibit high‐quality ablation with good channel effect at 50 mJ/mm2 and 30 Hz.

A Descriptive Study: Evaluation of Delayed Laboratory Testing in Emergency Room

Precision, accuracy, and short turnaround time (TAT) are important to improve the quality and effectiveness of laboratory services in the Emergency Room (ER). Several studies reported an average of 60% the ratio of anticoagulant to sample is not appropriate, lysis, and clotted samples occurred in the pre-analytical part. This part consisted of the request/registration of laboratory tests, patient preparation, specimen collection, and transportation to the central laboratory. The aim of this research is to evaluate the cause of the long TAT for laboratory research so as to obtain problem solving based on the target. This observational study was conducted in 2021 at the ER laboratory outlet. Observations were made started from the workload carried out by the laboratory technician, sample registration, and data collection from the Laboratory Information System (LIS). The study obtained 14.709 samples from the LIS at the Clinical Pathology Department, Cipto Mangunkusumo Hospital. The highest sample rejection rate was from the ER laboratory outlet due to specimen lysis (2.79%). The average time required for re-collecting specimens was around 2-2.5 hours. This rejection was below the minimum quality standards based on the International Federation of Clinical Chemistry (IFCC). Other problems include the limited number of medical laboratory experts compared to the workload also inefficient Computerized Physician Order Entry (CPOE) registration methods. The main problem of delayed laboratory testing in the emergency department is the pre-analytical part including sample registration and rejection specimens. That rejection mostly comes from lysis specimens, which caused invalid laboratory result.

Preanalytical Errors in Hematology: Insights From a Tertiary Care Hospital.

Introduction Errors occur in the laboratory at any level of the testing process. Recognizing these errors may cause patient distress by delaying diagnosis and management until results are released. This study aims to assess preanalytical errors in the laboratory and suggests methods to prevent them to improve accuracy and efficiency in the hematology department of a tertiary care hospital. The background emphasizes the critical role of preanalytical procedures in ensuring accurate hematological diagnoses and highlights the frequent mistakes that occur during specimen collection, handling, and transportation. Staff training, process standardization, using suitable collecting and transport equipment, putting quality control measures in place, and using automation technologies are some strategies for reducing errors and speeding up turnaround times. To increase diagnostic accuracy, patient care outcomes, and laboratory efficiency, hematology laboratories should address preanalytical mistakes and employ fostering methods. Aims and objectives The objective of this research is to evaluate the frequency and characteristics of preanalytical errors within the hematology laboratory of a tertiary care hospital. The study seeks to gauge the scope of the problem, identify key factors contributing to these errors, such as issues in specimen collection, handling, and transportation, and highlight areas where improvements can be made. Also, it intends to assess how preanalytical errors affect hematological accuracy and turnaround times, with a focus on patient care outcomes, and recommend techniques to reduce preanalytical errors and improve the accuracy of hematological results. Materials and methods This study was conducted at the hematology laboratory of our hospital from January 2023 to June 2024 after getting proper approval from the Institutional Review Board (IRB approval number 294/08/2024/PG/SRB/SMCH). It is a retrospective analytical study, and the study population comprised samples of patients from the emergency, inpatient, and outpatient departments, which included 51,155 complete blood count (CBC) and 5,449 peripheral smear(PS) samples. The study included only test samples sent specifically for hematological analysis, while those sent for cytological, biochemical, or microbiological testing were excluded. The distribution frequency of the samples and the number of rejected samples were analyzed, and the results were then compared and correlated. Results During the study period, a total of 56,604 samples were processed in the hematology laboratory. Of these, 886 samples (1.3%) were rejected due to preanalytical errors. The most frequent error was submitting an insufficient sample (54.17%), while the least frequent was the use of an empty or defective tube (0.4%). In the emergency department, the primary issues were insufficient and clotted samples, while in pediatric cases, errors were mainly due to inadequate or diluted samples. Conclusion Preanalytical errors in hematology laboratories, though often overlooked, can significantly impact diagnostic accuracy and patient care. Our study highlights that a substantial portion of errors arise from inadequate sample collection and handling, particularly in emergency and pediatric cases. Adherence to standard laboratory techniques can dramatically reduce preanalytical errors.

Using clotted, pelleted blood samples for direct molecular detection of Bartonella spp. in small mammal wildlife surveillance studies

ObjectiveBartonella are emerging bacterial zoonotic pathogens. Utilization of clotted blood samples for surveillance of these bacteria in wildlife has begun to supersede the use of tissues; however, the efficacy of these samples has not been fully investigated. Our objective was to compare the efficacy of spleen and blood samples for DNA extraction and direct detection of Bartonella spp. via qPCR. In addition, we present a protocol for improved DNA extraction from clotted, pelleted (i.e., centrifuged) blood samples obtained from wild small mammals.ResultsDNA concentrations from kit-extracted blood clot samples were low and A260/A280 absorbance ratios indicated high impurity. Kit-based DNA extraction of spleen samples was efficient and produced ample DNA concentrations of good quality. We developed an in-house extraction method for the blood clots which resulted in apposite DNA quality when compared to spleen samples extracted via MagMAX DNA Ultra 2.0 kit. We detected Bartonella in 9/30 (30.0%) kit-extracted spleen DNA samples and 11/30 (36.7%) in-house-extracted blood clot samples using PCR. Our results suggest that kit-based methods may be less suitable for DNA extraction from blood clots, and that blood clot samples may be superior to tissues for Bartonella detection.

Ultrafast intraoperative parathyroid hormone monitoring system: prospective, multicentre, clinical validity study.

Intraoperative parathyroid hormone (PTH) monitoring is a proven and reliable adjunct to parathyroid surgery, able to improve the outcomes and efficiency of the diagnostic and therapeutic pathway for patients with primary hyperparathyroidism. This study evaluated the innovative, compact, fully automated NBCL CONNECT Analyzer, which can measure whole-blood PTH in 5 min. A prospective multicentre study was conducted in stages: results reviews, recommendations, and implementation of improvements to the mechanical design, components of cartridges, calibration, and sampling protocols. Patients undergoing parathyroidectomy had PTH levels measured on the Analyzer and main laboratory platforms, either Roche or Abbott. The Miami criterion of a 50% drop in PTH concentration was used to define biochemical cure during surgery, and normal postoperative calcium level as cure of primary hyperparathyroidism. Measurements on the Analyzer were done by laboratory staff in London and nurses in Stuttgart. The Pearson coefficient (R) and Wilcoxon test were used for statistical analysis. Some 234 patients (55 male, 179 female) with a median age of 58.5 (age full range 15-88) years underwent parathyroidectomy (195 minimally invasive, 38 bilateral neck exploration, 1 thoracoscopic; 12 conversions) for primary hyperparathyroidism between November 2021 and July 2022. Primary hyperparathyroidism was cured in 225 patients (96.2%). The sensitivity, specificity, and overall accuracy of the Analyzer assay in predicting biochemical cure were 83.9, 100, and 84.8% in phase 1; 91.2, 100, and 91.3% in phase 2; and 98.6, 100, and 98.6% in phase 3. There were no false-positive results (positive predictive value 100%). Correlations between Analyzer measurements and those obtained using the Roche device were very strong (R = 0.98, P < 0.001 in phase 1; R = 0.92, P < 0.001 in phase 2; R = 0.94, P < 0.001 in phase 3), and correlations for Analyzer readings versus those from the Abbott platform were strong (R = 0.82, P < 0.001; R = 0.89, P < 0.001; R = 0.91, P < 0.001). The Analyzer showed continued good mechanical performance, with stable and repeatable operations (calibrations, quality controls). Introducing a stricter sampling protocol and improvements in the clot-detecting system led to a decrease in the number of clotted samples and false-negative results. Outcomes were not affected by measurements performed either by nurses or laboratory staff. Intraoperative PTH monitoring during parathyroid surgery can be done accurately, simply, and quickly in whole blood using the Analyzer.

Elastic Properties of Aging Human Hematoma Model In Vitro and Its Susceptibility to Histotripsy Liquefaction

ObjectiveTissue susceptibility to histotripsy disintegration has been reported to depend on its elastic properties. This work was aimed at investigation of histotripsy efficiency for liquefaction of human hematomas, depending on their stiffness and degree of retraction over time (0–10 d). MethodsAs an in vitro hematoma model, anticoagulated human blood samples (200 mL) were recalcified at different temperatures. In one set of samples, the shear modulus was measured by shear wave elastography during blood clotting at 10℃, 22℃ and 37℃, and then daily during further aging. The ultrastructure of the samples was analyzed daily with scanning electron microscopy (SEM). Another set of blood samples (50–200 mL) were recalcified at 37℃ for density and retraction measurements over aging and exposed to histotripsy at varying time points. Boiling histotripsy (2.5 ms pulses) and hybrid histotripsy (0.2 ms pulses) exposures (2 MHz, 1% dc, P+/P–/As = 182/–27/207 MPa in situ) were used to produce either individual cigar-shaped or volumetric (0.8–3 mL) lesions in samples incubated for 3 h, 5 d and 10 d. The obtained lesions were sized, then the lysate aspirated under B-mode guidance was analyzed ultrastructurally and diluted in distilled water for sizing of residual fragments. ResultsIt was found that clotting time decreased from 113 to 25 min with the increase in blood temperature from 10℃ to 37℃. The shear modulus increased to 0.53 ± 0.17 kPa during clotting and remained constant within 8 d of incubation at 2℃. Sample volumes decreased by 57% because of retraction within 10 d. SEM revealed significant echinocytosis but unchanged ultrastructure of the fibrin meshwork. Liquefaction rate and lesion dimensions produced with the same histotripsy protocols correlated with the increase in the degree of retraction and were lower in retracted samples versus freshly clotted samples. More than 80% of residual fibrin fragments after histotripsy treatment were shorter than 150 µm; the maximum length was 208 µm, allowing for unobstructed aspiration of the lysate with most clinically used needles. ConclusionThe results indicate that hematoma susceptibility to histotripsy liquefaction is not entirely determined by its stiffness, and correlates with the retraction degree.

Pre-Analytical Error in Biochemistry Laboratory

An observational study was done for a period of 6 months from 1st January, 2023 to 30th June, 2023 in the clinical biochemistry laboratory at GCS Medical College, Hospital and Research Centre. The study delved into pre-analytical errors within clinical biochemistry laboratories, focusing on error types, their prevalence, and potential impact. The study aimed to identify and quantify errors occurring in the pre-analytical phase, spanning from sample collection to report generation. Among the recorded errors (n=50), the most frequent was insufficient sample volume, signifying a pressing concern. Another prevalent error was Tests Not Mentioned or Add-on Testing, accounting for 26% of all errors, potentially disrupting workflow. The research also highlighted additional errors, including hemolysis, clotted samples, contamination from infusion routes, and lipemic samples. The study underscored the significance of addressing these errors to ensure accurate and reliable test results, thereby enhancing patient care. Overall, it provided valuable insights into the landscape of pre-analytical errors in clinical biochemistry, emphasizing the need for improved procedures, enhanced training, and effective communication to enhance the quality and precision of laboratory testing and, ultimately, patient care.

Chagas Disease and Leishmaniasis, Sympatric Zoonoses Present in Human from Rural Communities of Venezuela.

Trypanosoma cruzi and Leishmania spp. coexist in several endemic areas, and there are few studies of Chagas disease and leishmaniasis coinfection worldwide; for this reason, the objective of this work was to determine the Chagas disease and leishmaniasis coinfection in several rural communities co-endemic for these diseases. A total of 1107 human samples from six co-endemic rural communities of Cojedes state, Venezuela, were analyzed. Serum samples were evaluated by ELISA, indirect hemagglutination, and indirect immunofluorescence for Chagas disease diagnosis, and individuals were evaluated for leishmaniasis by leishmanin skin test (LST). Approximately, 30% of the individuals were also analyzed by PCR (blood clot samples) for T. cruzi and for Leishmania spp. The 14.7% of the individuals were positive to Trypanosoma cruzi infection by serology, and 25.8% were positive to Leishmania spp. current or past infection by LST. Among the group with PCR results, 7.8% were positive for T. cruzi, and 9.4% for Leishmania spp. The coinfection T. cruzi/Leishmania spp. was 6.5%. The T. cruzi DTUs of the positive blood clot samples were TcI, revealed using the molecular markers: (i) intergenic region of the miniexon, (ii) D7 divergent domain of the 24Sα rDNA, (iii) size-variable domain of the 18S rDNA, and (iv) hsp60-PCR-RFLP (EcoRV). The Leishmania species identified were L. (Leishmania) mexicana and L. (Viannia) braziliensis. A high prevalence was found for T. cruzi and Leishmania spp. single and coinfections in almost all communities studied, being these results of relevance for the implementation of control programs in co-endemic areas.

Таксономическая характеристика бактериальной ДНК, выделенной из тромбов пациентов с ишемическим инсультом

There is evidence of the role of microorganisms in endothelial damage and, as a consequence of activation of blood coagulation pathways, thrombosis, which in turn is one of the risk factors for the development of acute cerebrovascular accidents (ACVA). Objective. To analyze the microbiological profile of DNA isolated from thrombus of patients with ACVA. Materials and methods. Fifty-seven thrombus samples obtained lifetime from the vessels of patients with ischemic-type ACVA during thrombectomy and thrombaspiration were selected for the study. All patients with ACVA were in the period of coronavirus infection reconvalescence. Twelve blood samples from patients without infectious diseases were used for comparison. Amplification of DNA isolated from biological material was performed using primers specific to the 16S rRNA gene fragment. The obtained PCR products were sequenced by Sanger sequencing method, with subsequent processing of chromatograms in the BCV package. RDP Classifier was used to determine the taxonomic affiliation of DNA sequences. Statistical analysis was performed in the R environment using Pearson's and Fisher's χ2 criteria. Test results were considered significant at p &lt; 0.05. Results. Using the RDP Classifier, the taxonomic affiliation of 16S rRNA gene fragment amplification products were determined for 43 blood and clot samples. DNA of Actinobacteria and Firmicutes taxa were most represented in the clot samples of patients with ACVA. Of the Firmicutes and Proteobacteria taxa, Firmicutes predominated in thrombi, whereas Proteobacteria predominated in blood. The obtained data are consistent with the data on the key role of systemic intestinal inflammation in the pathogenesis of ACVA. Conclusion. DNA fragments of Actinomycetales, Bacillales, and Clostridiales bacteria found in thrombi of patients with ACVA indicate that they can penetrate the damaged epithelial barrier and promote platelet aggregation and thrombus formation, increasing the risk of ischemic stroke. Key words: ischemic stroke, acute cerebral circulatory disorders, bacterial infection, RDP Classifier

Molecular Diagnosis of Scrub Typhus: Sample and Timing Matter

Objective: Scrub typhus (ST) is an acute febrile illness caused by Orientia tsutsugamushi. Laboratory tests are needed to confirm the diagnosis when the characteristic eschar is absent. This study aimed to evaluate the performance of 47 kDa qPCR using clotted and EDTA-treated blood among patients with suspected ST Methods: Clotted blood samples from 284 inpatients (IPs) and EDTA-treated blood samples from 194 outpatients (OPs) with suspected scrub typhus were collected from patients who were blood culture- and malaria-negative. ST IgM ELISA and 47 kDa qPCR were performed for detection of ST infections. Results: Among the IPs (n=284) and OPs (n=194), 41% and 63% were confirmed to have ST based on the case definition, respectively; the mortality rate was 1%. The 47 kDa qPCR sensitivity of the buffy coat (65.6%) was greater compared to clotted blood (37.1%). PCR was more likely to be positive in the 1st week of the illness; IgM ELISA positivity increased after the 5th day of the illness. Conclusion: The type of sample and the time of sample collection have a role in the diagnosis of ST. These preliminary results need to be confirmed by prospective multicentric studies.

Leukocyte- and Platelet-Rich Fibrin (L-PRF) Obtained from Smokers and Nonsmokers Shows a Similar Uniaxial Tensile Response In Vitro.

We evaluated and compared the biomechanical properties of Leukocyte-and Platelet Rich Fibrin L-PRF clots and membranes derived from smoker and nonsmoker donors. Twenty venous-blood donors (aged 18 to 50 years) were included after signing informed consent forms. L-PRF clots were analyzed and then compressed to obtain L-PRF membranes. L-PRF clot and membrane samples were tested in quasi-static uniaxial tension and the stress-stretch response was registered and characterized. Furthermore, scanning electron microscope representative images were taken to see the fibrin structure from both groups. The analysis of stress-stretch curves allowed us to evaluate the statistical significance in differences between smoker and nonsmoker groups. L-PRF membranes showed a stiffer response and higher tensile strength when compared to L-PRF clots. However, no statistically significant differences were found between samples from smokers and nonsmokers. With the limitations of our in vitro study, we can suggest that the tensile properties of L-PRF clots and membranes from the blood of smokers and nonsmokers are similar. More studies are necessary to fully characterize the effect of smoking on the biomechanical behavior of this platelet concentrate, to further encourage its use as an alternative to promote wound healing in smokers.

Особенности процесса ферментации молочно-соевой смеси

The plant milk market is actively developing at the moment, as many consumers suffer from lactose and casein intolerance, while vegetarianism is being popularized. The use of soy milk is becoming popular in the technology of fermented dairy plant products, as it is a lactose-free analog of cow milk and source of native protein, polyunsaturated fatty acids, vitamins, minerals, and other micronutrients. However, the fermentation of dairy and plant compositions is quite complex and has not been fully understood yet, and therefore the goal of this study was to conduct a comparative analysis of the fermentation of milk and soy mixtures with different compositions (cow milk: soy milk in the ratios of 55:45; 70:30; 85:15). The tests were conducted at the Laboratory of Agricultural Product Processing. To obtain milk and soy mixtures, a plant protein drink – soy milk – was prepared from soybeans of the Sentyabrinka variety selected by the Federal State Budgetary Scientific Institution Federal Research Center All-Russian Research Institute of Soybean. The fermentation of milk and soy mixtures was performed using the acid-rennet method with bacterial starter cultures No. 1 (Lactobacillus lactis, Lactococcus diacetylactis, Lactococcus cremoris, Streptococcus thermophilus) and No. 2 (Lactococcus lactis ssp. lactis, Lac. lactis subsp. diacetilactis, Leuconostoc lactis and/or mesenteroides subsp. cremoris, Lactobacillus plantarum, Lactobacillus helveticus, Streptococcus thermophilus). The acid formation of fermented milk and soy clots was studied in time at intervals of 1 hour, and their active and titratable acidity levels were analyzed in the course of work. Fermentation completion was determined based on the organoleptic indicators of fermented clots and their acidity levels. The following was established as the result of the tests: composition of the milk-soy mixture – 70:30; technological fermentation process parameters and modes – 328–346 minutes at a temperature of 34 °C, with the production of a fermented milk and soy clot having the best organoleptic quality indicators – well-formed, with a stable texture, pleasant, sour-milk taste, light soy aftertaste and smell. The analytical dependences of the titratable and active acidity indicators on the duration of fermentation process were calculated based on an analysis of acid formation in experimental clot samples.

First report of vancomycin resistant Enterococcus faecalis and Enterococcus faecium isolated from water buffalo clotted cream in Turkey

This study aimed to isolate the Enterococcus species from homemade water buffalo clotted cream sold in Afyonkarahisar, investigate the vancomycin resistance genes in isolated strains by PCR and determine the antibiotic resistance of strains to some antibiotics commonly used in Turkey. A total of 107 buffalo clotted cream samples sold in public bazaars of Afyonkarahisar were collected. Following the phenotypic identification using a commercial identification kit, PCR was applied to strains using by species specific primers. Forty Enterococcus strains were obtained from 107 samples by PCR. Of 40 strains, 31 (77.5%) and 9 (22.5%) were identified to be E. faecalis and E. faecium, respectively. One vanA (2.5%) and 14 vanB (35%) genes were determined in tested 40 strains. While of strains having the vanB gen, 10 and four were found to be E. faecalis and E. faecium, respectively, one E. faecium strain also harboured the vanA gen. Thus, while the vanB gen positivity was found to be 32.3% in 31 E. faecalis strains and 44.4% in 9 E. faecium strains, the vanA gen positivity was determined to be 11.1% in 9 E. faecium strains. It was not found the vanC1 and vanC2/C3 genes in none of the strains. While all of strains were phenotypically resistant to streptomycin and fusidic acid, high resistance rates were also determined in the strains to kanamycin (87.5%), cephalothin (80%), erythromycin (80%), gentamicin (77.5%), tetracycline (75%) and vancomycin (60%). Resistance to all tested antibiotics except teicoplanin was determined in E. faecalis strains and except chloramphenicol in E. faecium strains harbouring the vancomycin resistance genes. In conclusion, it was thought the buffalo clotted creams offered for consumption in Afyonkarahisar may have potential risk for public health in terms of VRE species. To our knowledge, this is the first study showing the presence of VRE in the clotted cream samples by PCR in Turkey.

Borrelia persica infection in wild carnivores in Israel: molecular characterization and new potential reservoirs

BackgroundBorrelia persica causes tick-borne relapsing fever in Israel, the eastern Mediterranean basin, and Asia. Relapsing fever is associated with severe illness and potentially death in humans and animals. Since B. persica infection has rarely been described in wild animals, the aim of this study was to evaluate the prevalence of infection with B. persica in wild carnivores in Israel.MethodsSpleen and blood clot samples from wild carnivores, which underwent necropsy, were tested for the presence of Borrelia DNA by real-time polymerase chain reaction (PCR). PCR products were sequenced, and the spirochete loads were quantified using a specific quantitative PCR (qPCR).ResultsA total of 140 samples from 74 wild carnivores were analyzed for the presence of Borrelia DNA. Six out of the 74 (8.1%) animals were found positive for B. persica by PCR and sequencing of the flagellin B gene, of which 4/74 (5.4%) were also positive by PCR for the glycerophosphodiester phosphodiesterase (glpQ) gene. Positive samples were obtained from three European badgers, and one striped hyena, golden jackal, and red fox each. All B. persica-positive animals were young males (P < 0.0001). Quantifiable results were obtained from 3/5 spleen and 4/5 blood samples. The spirochete loads in the blood were significantly higher than those found in the spleen (P = 0.034).ConclusionsThe prevalence of B. persica infection found in wild carnivores brought for necropsy was unexpectedly high, suggesting that this infection is widespread in some wild animal species in Israel. This is the first report of B. persica infection in the European badger and striped hyena. These carnivores have a wide geographical range of activity, and the results of this survey raise the possibility that they may serve as reservoir hosts for B. persica.Graphical

The Effect of Blood Clots on the Quality of RNA Extracted from PAXgene Blood RNA Tubes.

Background: PAXgene® Blood RNA tubes are routinely used in clinical research and molecular biology applications to preserve the stability of RNA in whole blood. However, in practice, blood clots are occasionally observed after blood collection and are often ignored. Currently, there are few studies on whether blood clots affect the quality of RNA extracted from these tubes. Materials and Methods: Fifteen pairs of non-clot and clot PAXgene Blood RNA tube samples (n = 30) were collected to form two matched groups from 15 patients. According to the maximum diameter (d) of the blood clot observed visually at the time of sample reception, the clot groups were divided into a small-clot group (0 cm < d < 0.5 cm) and a large-clot group (d ≥ 0.5 cm). RNA was extracted by the PAXgene Blood RNA Kit. To analyze the quality of RNA, its yield and purity were assessed by spectrophotometry, and integrity was measured by microfluidic electrophoresis. An A260/280 ratio between 1.8 and 2.2 indicated purified RNA, and RNA integrity number (RIN) values ≥7.0 were considered to represent qualified integrity. Results: The median yields of RNA from the non-clot and clot groups were 3.84 (2.80-6.38) μg and 4.87 (2.77-8.30) μg, respectively. The median A260/280 ratios were 2.08 (2.06-2.09) and 2.09 (2.07-2.11), whereas the median A260/230 ratios were 1.77 (1.31-1.91) and 1.67 (1.21-1.94) in the two groups. In addition, the median RINs were 8.20 (8.00-8.40) and 7.20 (6.60-7.70), respectively. There were no significant differences in RNA yields, A260/280, or A260/230 between the two groups. However, the RIN value of the clot group was significantly lower compared with the non-clot group (p < 0.05), with RIN ≥7.0 found in all non-clot samples and 60% of clot samples (p < 0.05). Furthermore, in the clot groups, the small-clot samples had higher RIN values than large-clot samples (8.25 [7.75-8.75] vs. 6.90 [6.60-7.30], p < 0.001). Conclusions: The formation of large blood clots in PAXgene Blood RNA tubes will reduce the integrity of extracted RNA.

Analysis of the Six Sigma Principle in Pre-analytical Quality for Hematological Specimens.

Introduction Blood tests are essential for detecting and treating hospitalized individuals with diseases. Laboratory blood tests provide doctors with critical information required to treat their patient's illnesses. The most common sources of error in clinical laboratories are pre-analytical errors. Although quality control measures can remediate analytical errors, there is a requirement for stringentquality checks in the pre-analytical sector as these activities are performed outside of the laboratory. Pre-analytical errors when combined with the sigma value can reflect a better pictureas the sigma value represents the laboratory's performance. Aim In this study, six sigma and the Pareto principle were utilizedto assess pre-analytical quality indicators for evaluating the performance of a clinical hematologylaboratory. Methodology This is a retrospective observational study conducted from 2015 to 2023 (for a period of eight years). Information about the frequency of pre-analyticalerrorswas retrieved from the hematology section of the central diagnostic researchlaboratory information system and the data was entered into an MS Excel sheet and datawas evaluated utilizingSPSS version 23(IBM Corp., Armonk, NY). Results In the current research, total of 15 pre-analytical errors were noted. Out of the total 15 pre-analytical errors studied, 55.4% of pre-analytical errors were notedamong which80% errors were due to lack of mention of sample type or received timeand20% of errorswere attributedto no mention of diagnosis in requisition forms. The next most common errors noted were insufficient samples (8.26%) followed by absence of physician's signature (7%), incomplete request form (5.4%), age (4.2%), unique hospital identification (UHID) number (3.7%), clotted samples and transportation of the samples (3.6%), date and incorrect vials (2.6%). Gender (0.95%), hemolysed(0.85%), and lipemic samples (0.45%).Hemolysed andlipemic samples hada sigma value of 4.4 and 4.6, respectively, whereas gender and age had a sigma value of 4.3 and 3.8, inadequate sample for testing and an incorrect anticoagulant to blood ratio had a sigma value of 3.6, indicating that sample collection has to beimproved as the inverse relationship is noted between sigma value and laboratory performance. Conclusion Pareto chart and sigma value can help recognize most common pre-analytical errors, which consequently will help to prevent further recurrence of pre-analytical errors. Adequate training with regard to best practices in phlebotomy for interns, clinicians and technicians must be provided to decrease quantitative errors, which will further enhancetotal quality management in the laboratory.

Clotted blood samples in the neonatal intensive care unit: A retrospective, observational study to evaluate interventions to reduce blood sample clotting.

Blood draws for laboratory investigations are essential for patient management in neonatal intensive care units (NICU). When blood samples clot before analysis, they are rejected, which delays treatment decisions and necessitates repeated sampling. To decrease the incidence of rejected blood samples taken for laboratory investigation as a result of clotted sample. This retrospective observational study used routine data on blood draws from preterm infants collected between January 2017 and June 2019 in a 112-cot NICU in Qatar. Quality improvement interventions to reduce the rate of clotted blood samples included: awareness raising and safe sampling workshops with NICU staff, involvement of the neonatal vascular access team, development of a complete blood count (CBC) sample collection pathway, review of sample collection equipment, introducing the Tenderfoot® heel lance, establishment of benchmarks and provision of dedicated blood extraction equipment. First attempt blood draw occurred in 10 706 cases, representing a 96.2% success rate. In 427 (3.8%) cases, the samples were clotted requiring repeat collection. The overall rate of clotted specimens decreased from 4.8% in 2017 and 2018 to 2.4% in 2019, with odds ratios of 1.42 (95% confidence interval [CI] 1.13-1.78, p = .002), 1.46 (95% CI 1.17-1.81, p < .001) and 0.49 (95% CI 0.39-0.63, p < .001), respectively. The majority (87%-95%) of blood samples were by venepuncture using an intravenous (IV) catheter or the NeoSafe™ blood sampling device. Heel prick sampling was the second (2%-9%) most common method. Clotted samples were most frequently associated with needle use, 228 of 427 (53%), and IV cannula, 162 of 427 (38%), with odds ratios of 4.14 (95% CI 3.34-5.13, p < .001) and 3.11 (95% CI 2.51-3.86, p < .001), respectively. Our interventions over 3 years were associated with reduced rates of sample rejection due to clotting, and this led to improved patient experience through fewer repeated samplings. The insights gained from this project can help to improve patient care. Interventions that reduce the rate of blood sample rejection by clinical laboratories can lead to economic savings, timelier diagnostic and treatment decisions, and contribute to an improved quality care experience for all critical care patients, irrespective of age, by reducing the need for repeated phlebotomy and the risk of related complications.

Frequency of Hypomagnesemia in Neonates presenting with Hypocalcemia in a Tertiary Care Hospital

Background: Hypocalcemia is a common neonatal occurrence, especially in neonates with high-risk including those babies born to diabetic mothers, neonates with perinatal asphyxia and premature babies. High calcium consumption can reduce magnesium absorption, while low magnesium intake can boost calcium absorption. Aim: To assess the frequency of hypo-magnesemia in neonates presenting with hypocalcemia in a tertiary care hospital. Design study: Cross sectional study. Methodology: Total 160 fulfilling the inclusion criteria at Services Hospital, Lahore were enrolled through non-probability convenience sampling. Gestational age, birth weight, gender and feeding pattern) were noted. Blood sample was obtained by using 3cc BD syringe and 20G butterfly needle with the help of a nurse under sterile measures. All clotted samples were stored and sent to the Pathology lab for the assessment of magnesium level. Reports were assessed and if magnesium level&lt;0.7mmol/L, then hypomagnesemia was labeled. Data was evaluated by using SPSS v.24. Quantitative data was presented as mean±SD. Qualitative data was presented as frequency and percentage. Chi-square test was applied with P-value ≤0.05 taken as significant. Results: In this study, there were 75 (46.9%) male and 85 (53.1%) female neonates. The mean gestational age of neonates was 39.20±1.63weeks. The mean birth weight of neonates was 2.5±3.0kg. There were 46 (28.8%) who were on exclusive breast feeding, 55 (34.4%) were on formula feed while 59 (36.9%) were on both; breast and formula feed. The mean magnesium level of neonates was 0.90±0.63mmol/L. There were 113(70.6%) neonates with hypomagnesemia while 47 (29.4%) had normal magnesium level. Practical Implication: Literature has reported that the frequency of hypomagnesemia is high in neonates with hypocalcemia. In Pakistan there is little local evidence available in this regard. The information gained through this study will help to improve our clinical practice and to update guidelines for early screening and management of infants with hypocalcemia with co-existence hypomagnesemia. Conclusion: It was concluded that the frequency of hypo-magnesemia was high in neonates with hypo-calcemia in local population. Keywords: Prevalence, Calcium, Electrolytes Imbalance and Neonates.

Preanalytical Errors in a Hematology Laboratory: An Experience from a Tertiary Care Center

Laboratory errors arise at any stage of testing. Detecting these inaccuracies before results are revealed might delay diagnosis and treatment, causing patient distress. Here, we studied the preanalytical errors in a hematology laboratory. This one-year retrospective analysis was conducted at the laboratory of a tertiary care hospital and included information on blood samples that were taken for hematology tests from both outpatients and inpatients. Laboratory records included sample collection and rejection information. The type and frequency of preanalytical errors were expressed as a proportion of total errors and sample number. Microsoft Excel was utilized to enter data. The results were presented in the form of frequency tables. This research included 67,892 hematology samples. For preanalytical errors, 886 samples (1.3%) were discarded. The most common preanalytical error was insufficient sample (54.17%), and the least common was an empty/damaged tube (0.4%). Erroneous samples in the emergency department were mostly insufficient and clotted, whereas pediatric sample errors were caused by insufficient and diluted samples. Inadequate samples and clotted samples account for the vast majority of preanalytical factors. Insufficiency and dilutional errors were most frequent from pediatric patients. Adherence to best laboratory practices can drastically cut down on preanalytical errors.