Coagulation Tubes Research Articles

Effect of using smaller blood volume tubes and closed blood collection devices on total blood loss in patients undergoing major cardiac and vascular surgery.

Diagnostic laboratory tests are an integral part of managing hospitalized patients. In particular, patients in the intensive care units (ICUs) can experience a concerning amount of blood loss due to diagnostic testing, which can increase the risk developing iatrogenic anemia. Several interventions exist to curtail avoidable blood loss, for example computerized decision support, smaller phlebotomy tubes, and other blood conservation devices. Nevertheless, use of these interventions is not standardized. Therefore, the objective of our study was to quantify the daily phlebotomy volume taken from patients who had undergone major cardiac or vascular surgery. We estimated the number of blood analyses and volumes of drawn blood of 400 consecutive patients (≥18 yr) undergoing major cardiac or vascular surgery. The amount of blood saved using small-volume tubes and in combination with blood conservation device rather than standard-volume tubes was estimated for serum chemistry (serum), ethylenediaminetetraacetic acid (EDTA) tubes, sodium citrate coagulation (SCC) tubes, and arterial blood gas (ABG) analysis. The mean total blood loss due to phlebotomy drawing using standard-volume tubes during hospitalization was 167.9mL (95% confidence interval [CI], 158.0 to 177.8), 255.6mL (95% CI, 226.5 to 284.6), and 695.3mL (95% CI, 544.1 to 846.4) for patients undergoing cardiac surgery with a hospital length of stay (LOS) of 0-10, 11-20, and ≥ 21 days, respectively. The mean total blood loss due to phlebotomy during hospitalization was 80.5mL (95% CI, 70.5 to 90.6), 225.0mL (95% CI, 135.1 to 314.8mL) and 470.3mL (95% CI, 333.5 to 607.1) for vascular surgery patients with LOS 0-10, 11-20, and ≥ 21 days, respectively. Patients with at least a two-day stay at the ICU had a mean blood loss of 146.6mL (95% CI, 134.6 to 158.6mL) and those with ≥ 11 days incurred a loss of 1,428mL (95% CI, 1,117.8 to 1,739.2). The use of closed blood collection device and small-volume tubes (serum, EDTA, SCC, and ABG) reduced blood loss by 82.8mL for patients with an ICU stay of 2 days and up to 824.0mL for patients with a ICU stay of ≥ 11 days. Diagnostic laboratory tests are associated with significant patient blood loss, but are a modifiable risk factor. The use of small-volume tubes and closed blood collection devices decreases the volume of patient blood drawn for analysis and prevents blood waste.

Are tube fill volumes below 90% a rejection criterion for all coagulation tests?

Rejected samples lead to prolonged turnaround time and delayed diagnosis and treatment of patients. This study was conducted to determine minimum acceptable sample volume in Sarstedt brand coagulation tubes to reduce high sample rejection rate. Blood samples were drawn from 20 participants (10 healthy volunteers and 10 patients receiving oral anticoagulant) into coagulation tubes. Six samples were taken from each participant, with tube fill volumes of 100%, 90%, 80%, 70%, 60%, and 50%. Prothrombin time (PT), active partial thromboplastin time (aPTT), and fibrinogen tests were analyzed. According to quality performance specifications, the tube fill volume must be at least 70% for PT and aPTT and 50% for fibrinogen. There was no statistical difference in samples from healthy volunteers for PT, aPTT, and fibrinogen tests when the minimum tube fill volume was at least 80%, 90%, and 50%, respectively. These percentages were 50%, 70%, and 60%, respectively, in patients receiving oral anticoagulant. Sarstedt tubes meet quality standard specifications at a 70% fill rate for PT and aPTT and a 50% fill rate for fibrinogen. Comprehensive studies with larger populations are needed to accept these values as sample acceptance criteria for the laboratory.

Change of routine coagulation parameters in plasma samples with different hematocrit values

Abstract Aim There are many preanalytical variables affecting routine coagulation tests. Increased hematocrit (Htc) levels are one of these variables. However, no study has been conducted to determine the effect of low Htc values on coagulation tests. Therefore, in this study, we aimed to evaluate whether low Htc values affect coagulation tests besides high Htc values. Methods Standard human plasma was injected into coagulation tubes containing 3.2% sodium citrate to reflect hematocrit rates of 5% to 75% and prothrombin time (PT), active partial thromboplastin time (aPTT), thrombin time (TT) and fibrinogen measurements were performed. Results Three groups were formed according to Htc levels: A (5-25%), B (30-50%) and C (55-75%). PT (s) were found 13.7±0.30 in group A, 14.88±0.57 s in group B, 20.16 ±4.66 s in group C respectively. aPTT (s) results were 35.79±1.39 s in group A, 42.48 ± 3.51 s in group B and 76.47 ± 31.55 in group C. TT (s) results were found to be 26.42 ± 0.77 s (group A), 28.24±1.17 s (group B) and 32.02±2.60 (group C). Fibrinogen levels (g/L) were measured as 2.30 ± 0,05, 2.21 ± 0,07 and 1.90 ± 0.20 in groups A, B, C, respectively. For all measured parameters, group A reflecting low Htc was significantly (P < 0 ,0001) different from the other groups. Conclusion Previous studies have reported that high Htc (> 55%) levels affected routine coagulation tests. In our study, low Htc (5-25%) values were also shown to cause errors in the test results.

Phenotypic characteristic of Staphylococcus aureus from subclinical mastitis in Etawah-crossbreed goats in Yogyakarta, Indonesia.

Subclinical mastitis (SCM) in Etawah-grade (PE) goats in Yogyakarta, Indonesia, is commonly due to Staphylococcus aureus. At present, S. aureus from SCM in PE goats in Yogyakarta has not been characterized. Therefore, this study aimed to phenotypically characterize S. aureus, which has been isolated from SCM of PE goats. A total of 314 lactating PE goats were collected from 60 PE goat farms (e.g., Sleman, Bantul, and Kulonprogo) located in parts of Yogyakarta with an average age of 3-4 years old, three of which showed SCM based on the California mastitis test (CMT). Subclinical mastitis is confirmed in PE goats if CMT shows ++ or +++. Furthermore, S. aureus was detected by biochemical assays. Staphylococcus aureus could determine hemolysin (Hae), coagulase (Coa), clumping factor (Cf), and antibiotic susceptibility. Hemolytic bacteria were detected by culturing on blood agar plate, and Cf was detected by slide agglutination. The production of Coa was detected by tube coagulation. Staphylococcus aureus susceptibility was determined by antimicrobial agar diffusion using a paper disc. Phenotypically characterized S. aureus from PE goats with SCM in Yogyakarta, Indonesia, Coa-, Cf-, and Hae- were found to be resistant to erythromycin (ERYTHRO), ampicillin (AMP), penicillin (PEN-G), and sulfamethoxazole (SULFA). The phenotypic characteristic of S. aureus, which was obtained from SCM in PE goats in Yogyakarta, consists of Coa, and Cf-. S. aureus cannot perform hemolysis of red blood cells. This phenotypic characteristic can prevent and control SCM in PE goats. Several antibiotics such as ERYTHRO, AMP, PEN-G, and SULFA were no longer effective for treating SCM in PE goats because S. aureus has developed its resistance to these antibiotics.

A Comparative Study of Biochemical Markers in the Early Diagnosis of Acute Myocardial Infarction (AMI)

Background: One of the most effective instances of readily detected biomarkers diagnosing a very big health concern is the measuring of proteins in blood to represent heart damage. The idea of utilizing a blood test to reflect organ or cell harm necessitates a chemical that is plentiful in the target cell, can reach blood, has a suitable half-life in blood, and is preferably in a particular form that only reflects the target cell in tissue. Because contraction is the primary function of the myocyte, proteins involved in contraction or the energy required to sustain it should be suitable potential indicators. The work on diagnostic tests for myofibrillar proteins begins in 1978. Different key pieces of evidence fueled in the research. In Ist-year outcomes were identical in chest pain patients whose acute myocardial infarction (AMI) was ruled out by electrocardiography and cardiac enzymes, demonstrating that the existing diagnostic tools were not beneficial for risk prediction and treatment recommendations. Second, CK and lactate dehydrogenase isoenzymes were brought into clinical practice and demonstrated to have higher specificity and sensitivity than traditional tests. All of the biomarkers used to diagnose cardiac injury are involved in contraction or energy metabolism, but the markers evolved via trial and error, beginning with transaminases in the 1950s and ending with troponins in the 1990s. This background is discussed, along with observations on my experiences establishing CK-MB and Troponin I assays. Methods: 140 individuals with AMI are included who are diagnosed using WHO criteria. A cross-sectional study with purposive sampling technique is conducted in wazirabad institute of Cardiology. This research is last around 6 months. Before the examinations, blood samples are obtained from patients by vein puncture using syringes and preserved in clot tubes. The research includes patients experiencing chest pressure, tightness, or discomfort, shortness of breath, and irregular cardiac rhythms on ECG. Patients having a normal ECG are not included in the study. Different equipments are utilized to assess appropriate heart function and to obtain serum for the identification of various cardiac biomarkers. Result: The sensitivities of all biochemical markers change depending on the time of infarction, as previously demonstrated and expected from a patho-physiologic standpoint, the blood sample of all the patients was collected and used for further investigation in the Lab. In the current study, 140 patients are enrolled, and all of the patients who had a single episode of AMI had their ECGs taken. All of the patients' ECGs are abnormal. In compared to other Bio-Cardiac Markers, the Troponin –I has a higher ratio of patients, according to the present findings. When a person undergoes AMI, the first protein that is released is troponin-I. When these tests are conducted over a period of time, they provide us with a quick assessment of the severity of AMI. As a result of the current findings, patients have a higher level of Trop-I in their blood when they experience their first episode of AMI. In this situation, the patient should go to the nearest health care center's emergency room as quickly as feasible. Conclusion: The traditional enzymatic assays of creatine kinase (CK), CK-MB (activity), and lactate dehydrogenase are rapidly being replaced by mass measurements of myoglobin, CKMB, and troponin T and troponin I (Tnl) 1 " to achieve high diagnostic sensitivity and specificity within a few hours from the onset of tissue necrosis; the ultimate goal is the early and appropriate management of patients' conditions. We compared the diagnostic sensitivity and specificity obtained by measuring various biochemical markers, some of which are widely used, and concluded that Troponin I is the first cardiac marker that releases simultaneously after the first episode of AMI when performed as single tests and in parallel and serial modes.

Negligible Specimen Hemolysis Is Observed With Sarstedt S-Monovette Coagulation Tubes in Aspiration Mode.

We aimed to compare the levels of hemolysis in the blood collected using the vacuum and aspiration modes via Sarstedt S-Monovette coagulation tubes. Forty volunteers were included in the study. Blood samples were collected using two different modes in the S-Monovette citrate tube (Sarstedt AG). Prothrombin time, active partial thromboplastin time, fibrinogen, and D-dimer analyses were performed using the STA-Compact-Max 3 analyzer (Stago). The hemolysis levels of the samples were measured by both Stago's semiquantitative hemolysis index (H-index) module and the quantitative H-index measurement of the Roche cobas 6000 (Roche Diagnostics) analyzer. Roche's quantitative H-index values were statistically significantly lower in the aspiration mode. No clinically significant difference was observed between coagulation test results. Using the S-Monovette citrate tubes can reduce spurious hemolysis and improve patient safety.

The Effect of Bacterial Infection with Helicobacter pylori on Thyroid Hormones

Objective: Our current research aimed to see if there was a link between Helicobacter pylori infection and thyroid hormone levels. Methods: In Karbala, Iraq, samples were obtained from 84 individuals with H. pylori (42 patients tested positive for H. pylori antigen test and 42 patients tested negative) between December 2021 and January 2022. All subjects had five milliliters of whole blood drawn into clot tubes and centrifuged for 30 minutes to obtain serum. Enzyme-linked immunosorbent assay (ELISA) kits were used to measure the amounts of free T4 and TSH in the blood. Results: Females are more susceptible to H. pylori infection than males (p = 0.00001), according to the findings. Furthermore, the mean age of positive test patients is 49.66 years, which is considerably (p = 0.0177) higher than the mean age of negative test patients (43.38 years), and the oldest men are more infected with H. pylori than young men. The results show no significant variations in free T4 concentrations between males and females for both positive and negative tests (p > 0.05). This implies that while infected with H. pylori, the concentration of free T4 was unaffected. Finally, H. pylori infection causes a considerable increase in stomach acid, according to the research. (p = 0.0037) in FSH levels in both sexes. Conclusions: We can determine that females are more likely than males to be infected with H. pylori. And that most patients are aged, particularly men. Furthermore, there is no partnership between H. pylori infection and free F4 hormone levels, and patients with H. pylori have a higher FSH hormone concentration than those with a negative H. pylori antigen test.

M235 Contradiction between separation gel coagulation tube and heparin anticoagulation tube

M235 Contradiction between separation gel coagulation tube and heparin anticoagulation tube

Analysis of the Increase of Vascular Cell Adhesion Molecule-1 (VCAM-1) Expression and the Effect of Exposure in a Hyperbaric Chamber on VCAM-1 in Human Blood Serum: A Cross-Sectional Study.

Background and Objectives: Vascular cell adhesion molecule-1 (VCAM-1) was identified as a cell adhesion molecule that helps to regulate inflammation-associated vascular adhesion and the transendothelial migration of leukocytes, such as macrophages and T cells. VCAM-1 is expressed by the vascular system and can be induced by reactive oxygen species, interleukin 1 beta (IL-1β) or tumor necrosis factor alpha (TNFα), which are produced by many cell types. The newest data suggest that VCAM-1 is associated with the progression of numerous immunological disorders, such as rheumatoid arthritis, asthma, transplant rejection and cancer. The aim of this study was to analyze the increase in VCAM-1 expression and the impact of exposure in a hyperbaric chamber to VCAM-1 levels in human blood serum. Materials and Methods: The study included 92 volunteers. Blood for the tests was taken in the morning, from the basilic vein of fasting individuals, in accordance with the applicable procedure for blood collection for morphological tests. In both groups of volunteers, blood was collected before and after exposure, in heparinized tubes to obtain plasma and hemolysate, and in clot tubes to obtain serum. The level of VCAM-1 was determined using the immunoenzymatic ELISA method. Results: The study showed that the difference between the distribution of VCAM-1 before and after exposure corresponding to diving at a depth of 30 m was at the limit of statistical significance in the divers group and that, in most people, VCAM-1 was higher after exposure. Diving to a greater depth had a much more pronounced impact on changes in VCAM-1 values, as the changes observed in the VCAM-1 level as a result of diving to a depth of 60 m were statistically highly significant (p = 0.0002). The study showed an increase in VCAM-1 in relation to the baseline value, which reached as much as 80%, i.e., VCAM-1 after diving was almost twice as high in some people. There were statistically significant differences between the results obtained after exposure to diving conditions at a depth of 60 m and the values measured for the non-divers group. The leukocyte level increased statistically after exposure to 60 m. In contrast, hemoglobin levels decreased in most divers after exposure to diving at a depth of 30 m (p = 0.0098). Conclusions: Exposure in the hyperbaric chamber had an effect on serum VCAM-1 in the divers group and non-divers group. There is a correlation between the tested morphological parameters and the VCAM-1 level before and after exposure in the divers group and the non-divers group. Exposure may result in activation of the endothelium.

Cinelaparoscopy - the Remarkable Early Progress in Video-Documentation of Laparoscopic Surgery.

Study Objective Historical presentation of Cinelaparoscopy to document surgery and educate physicians and patients in minimally invasive gynecologic surgery (MIGS). Design Presentation of an award-winning paper on the utilization of the technique in MIGS. Setting Armed Forces Hospital. Patients or Participants Case reports of gynecologic patients opting for minimally invasive gynecologic surgery. Interventions Developing and utilizing the equipment, including 8mm cameras, to document laparoscopic gynecologic surgery. Measurements and Main Results An 8mm movie camera was attached to a laparoscope to document gynecologic procedures including ovarian biopsy, retrieval of an intrauterine device (IUD) that had perforated the uterus, monopolar Fallopian tube coagulation for sterilization and survey of the pelvis, appendix, intestines, liver and gall bladder. After the film was developed, it was edited and spliced for presentation at a national meeting of obstetricians and gynecologists. One can imagine the excitement that the pioneers in MIGS experienced when seeing these films. Metaphorically, the world of MIGS experienced "Moore's Law" in the 1970′s with the rapid development of safer techniques, better instrumentation, bipolar coagulation devices, improved optics, and innovative performances of standard gynecologic procedures, e.g., Laparoscopic Hysterectomy. Conclusion This award-winning presentation was instrumental in the planning, the documentation, and the distribution of the 1976 and 1977 Hawai'i AAGL endoscopic teaching videos. Those videos increased the awareness of MIGS, spurred the development of MIGS procedures, and encouraged the promulgation of MIGS as the preferred route of most gynecologic surgery. Historical presentation of Cinelaparoscopy to document surgery and educate physicians and patients in minimally invasive gynecologic surgery (MIGS). Presentation of an award-winning paper on the utilization of the technique in MIGS. Armed Forces Hospital. Case reports of gynecologic patients opting for minimally invasive gynecologic surgery. Developing and utilizing the equipment, including 8mm cameras, to document laparoscopic gynecologic surgery. An 8mm movie camera was attached to a laparoscope to document gynecologic procedures including ovarian biopsy, retrieval of an intrauterine device (IUD) that had perforated the uterus, monopolar Fallopian tube coagulation for sterilization and survey of the pelvis, appendix, intestines, liver and gall bladder. After the film was developed, it was edited and spliced for presentation at a national meeting of obstetricians and gynecologists. One can imagine the excitement that the pioneers in MIGS experienced when seeing these films. Metaphorically, the world of MIGS experienced "Moore's Law" in the 1970′s with the rapid development of safer techniques, better instrumentation, bipolar coagulation devices, improved optics, and innovative performances of standard gynecologic procedures, e.g., Laparoscopic Hysterectomy. This award-winning presentation was instrumental in the planning, the documentation, and the distribution of the 1976 and 1977 Hawai'i AAGL endoscopic teaching videos. Those videos increased the awareness of MIGS, spurred the development of MIGS procedures, and encouraged the promulgation of MIGS as the preferred route of most gynecologic surgery.

High preanalytical non-compliance and samples rejection rate in clinical biochemistry laboratory are decreased by nurse staff training in phlebotomy and sample handling

Background: Preanalytical phase of biomedical analysis remains an important source of diagnostic errors that deserves special attention. This study aims to evaluate the training in phlebotomy and sample handling impact on the preanalytical non-compliances. Material and Methods: we performed a prospective study before and after staff training in phlebotomy and sample handling by systematically recording all clinical samples non-compliances. First, we assessed and describe the non-compliance baseline rate from January to December 2017 in the clinical biochemistry laboratory of Hôpital Sominé DOLO de Mopti. After two sessions of one week staff training in January 2018, we performed the same study from January to December 2018. We compared the proportions of non-compliances between the two assessments. Data were collected on the case report forms, captured in Excel and analyzed by R software for (Mac) OS X version 4.0.3. Pearson Ch2 or Fisher exact tests were used for the comparison of proportions. The statistical significance was set at p < 5%. Results: a total of 27,810 venous blood samples were received during the study period; 48% was for biochemistry, 41% for immuno-serology, 9% for blood cell count and 2% for coagulation tests. There were 3,826 instances of preanalytical non-compliances (13.76%) identified that led to sample rejection. Out of the 11 types of non-compliances investigated, 5 (45.4%) accounted for nearly 91% of the problems: insufficient sample volume (28.9%), hemolyzed samples (20.5%), inappropriate collection time (17.8%), sample clot (12.9%), and inappropriate sample collection tube (10.8%). We observed a significant difference in rates of non-compliance between inpatients and outpatients samples (44.4% vs 7.3%; p < 0.001). The proportion of non-compliance have significatively decreased after the two training sessions of hospital staff in phlebotomy and sample handling 3,826/27,810 (13.8%) vs 3,009/32,476 (9.3%); p < 0.001. Conclusion: we report a significantly higher rate of non-compliance in inpatients. Hospital staff training in phlebotomy and sample handling reduce the proportion of preanalytical non-compliance and thereby improve patient management and safety.

Einfluss einer maßgeschneiderten Präanalytik-Fortbildung auf die Qualität der Laboruntersuchungen

In 31 to 75 percent of cases, errors in laboratory medicine have preanalytical causes such as erroneous blood sampling. Erroneous blood sampling may lead to false test results and additional laboratory cost; it may increase analyzing time and endanger the health of patients and employees. In particular, under- and overfilling of blood sampling tubes can considerably distort laboratory values. So far there has been a lack of studies investigating the effect of a tailored training for ward staff to improve preanalytical procedures on blood sampling. Can a tailored preanalytical training significantly reduce the number of commented under- and overfilled coagulation samples, reduce the number of hemolytic serum and lithium heparin samples and increase the number of standards-compliant blood sampling? In an intervention study we compared the number of commented under- and overfilled coagulation tubes and the number of hemolytic serum and lithium heparin samples on the basis of laboratory data and, using participant observation, compared the blood sampling quality on a surgical ward before and after participation in a training course. Based on prior results of participant observation, a 20-minute training was conceptualized and conducted. Target criteria were a) the number of commented under- and overfilled coagulation tubes and b) the number of hemolytic serum and lithium heparin samples in a before/after comparison (Oct-Dec 2017 and Jan-Mar 2018 compared with Jun-Aug 2018) and an annual comparison (Jun-Aug 2017 compared to Jun-Aug 2018), and c) a standards-compliant performance of blood sampling in a before/after comparison (Apr 2018 and Jun 2018). The number of commented under- and overfilled coagulation tubes in the annual comparison was compared using the Chi-square test. After the training (Jun-Aug 2018) the number of commented under- and overfilled coagulation tubes decreased significantly in annual comparison to Jun-Aug 2017 (-68.07%, p<0.001). The number of commented under- and overfilled coagulation tubes and of hemolytic serum and lithium heparin samples decreased in the before/after and in the annual comparison, and the number of standards-compliant blood samplings increased in the before/after comparison. The training contributed significantly to reducing the number of commented coagulation samples and hemolytic serum and lithium heparin samples and to increasing standards-compliant blood sampling. There is a need to investigate to what extent this concept can be transferred to other wards, and in which intervals further trainings should be conducted in order to maintain these positive effects.

Novel Thermostable Inhibitor of Contact Activation Tica Effectively Blocks Contact Activation in Low Tissue Factor Thrombin Generation

Background: Thrombin generation and other clotting assays suffer from a wide variation of pre-analytical variables. One of those pre-analytical variables is contact activation through blood withdrawal methods, different syringes, differences in blood coagulation tubes, blood transport and sample handling. It has been shown that the addition of contact activation inhibitors in low tissue factor activated thrombin generation leads to a correction of the, in these circumstances significant, increase in thrombin generation due to contact activation. We compare the novel 'thermostable inhibitor of contact activation' (TICA) to the current standard 'corn trypsin inhibitor' (CTI). Aim: Comparing the effectiveness of novel contact activation inhibitor TICA to the current standard CTI in low tissue factor-induced thrombin generation and recalcification in sodium citrate anticoagulated platelet poor plasma (PPP) and platelet rich plasma (PRP). Methods: We compared TICA, Corn trypsin inhibitor and plasma without contact activation inhibitors in low tissue factor PPP thrombin generation and in PRP recalcification thrombin generation, the latter the most sensitive condition for contact activation. In addition, we compared low tissue factor activated thrombin generation in plasma from severe hemophilia A patients with and without TICA during and after blood drawing. Thermostability - as a measure of shelf life - was measured and compared to CTI. Results: TICA is able to fully block contact activation in PRP recalcification experiments and is comparable to CTI in doing so. TICA significantly lowers low tissue factor induced thrombin generation by blocking contact activation. Pre-loading vacuum blood collection tubes with contact activation inhibitors is superior in inhibiting contact activation compared to addition of the inhibitor during the thrombin generation assay itself. TICA did not alter coagulation activity when added to FXIIa deficient plasma in thrombin generation. In contrast to CTI TICA is heat stable which will be of benefit to shelf life of pre-loaded blood drawing tubes. Conclusion: TICA is able to fully block contact activation and has several advantages over CTI. Disclosures No relevant conflicts of interest to declare.

Etiology and Antimicroba Susceptibility of Udder Pathogens from Cases of Subclinical Mastitis in Dairy Ettawa Crosbread Goat (PE) in Kulonprogo Yogyakarta

Subclinical mastitis in Ettawa crosbreeds (PE) is an inflammatory disease that no clinical symptoms, but there is an increase the number of somatic cells and causes decrease milk production which economically detrimental. The aim of these study was to isolation of bacteria that causing subclinical mastitis in PE goats and their susceptibility with antimicrobial. A total of 37 PE goats from 5 farms in Kulonprogo were tested by California Mastitis Test (CMT). PE goats were said subclinical mastitis if the CMT test positive (++) or (++). Isolation and identification bacteria from PE goat subclinical mastitis by bacteriological analytical manual. Coagulase test with tube coagulation and antimicroba susceptibility with agar diffusion. Subclinical mastitis in PE goats at Kulonprogo farm is 10/37 (27%) which caused by S. intermedius positive coagulase 4/4 (100%), S. aureus negative coagulase 7/10 (70%), S. aureus positive coagulase 3/10 (30%), and E. coli 1/10 (10%). S. intermedius positive coagulase was resistant to ampicillin, tetracycline, and sulfamethoxazole 2/4 (50%) respectively. S. aureus negative coagulase was resistant to ampicillin 2/7 (28.6%), penicillin 1/7 (14.3%), and sulfamethoxazole 1/7 (14.3%). S. aureus positive coagulase was resistant to ampicillin 1/3 (33.3%), penicillin 3/3 (100%), sulfametoxazole 2/3 (66.7%), and tetracycline group 1/3 (33.3%). Subclinical mastitis PE goats in Kulonprogo was caused by S. intermedius positive coagulase and S. aureus negative coagulase which resistant to penicillin and sulfamethoxazole.

Pre-analytical practices for routine coagulation tests in European laboratories. A collaborative study from the European Organisation for External Quality Assurance Providers in Laboratory Medicine (EQALM).

Background Correct handling and storage of blood samples for coagulation tests are important to assure correct diagnosis and monitoring. The aim of this study was to assess the pre-analytical practices for routine coagulation testing in European laboratories. Methods In 2013-2014, European laboratories were invited to fill in a questionnaire addressing pre-analytical requirements regarding tube fill volume, citrate concentration, sample stability, centrifugation and storage conditions for routine coagulation testing (activated partial thromboplastin time [APTT], prothrombin time in seconds [PT-sec] and as international normalised ratio [PT-INR] and fibrinogen). Results A total of 662 laboratories from 28 different countries responded. The recommended 3.2% (105-109 mmol/L) citrate tubes are used by 74% of the laboratories. Tube fill volumes ≥90% were required by 73%-76% of the laboratories, depending upon the coagulation test and tube size. The variation in centrifugation force and duration was large (median 2500 g [10- and 90-percentiles 1500 and 4000] and 10 min [5 and 15], respectively). Large variations were also seen in the accepted storage time for different tests and sample materials, for example, for citrated blood at room temperature the accepted storage time ranged from 0.5-72 h and 0.5-189 h for PT-INR and fibrinogen, respectively. If the storage time or the tube fill requirements are not fulfilled, 72% and 84% of the respondents, respectively, would reject the samples. Conclusions There was a large variation in pre-analytical practices for routine coagulation testing in European laboratories, especially for centrifugation conditions and storage time requirements.

Filling accuracy and imprecision of commercial evacuated sodium citrate coagulation tubes

Current recommendations advocate that blood tubes for coagulation testing should be filled not less than 90% of their nominal filling volume, since under- or over-filling >10% may generate unreliable results of some hemostasis assays. This study was hence aimed to explore filling accuracy and precision of commercial blood tubes. Between-lot variations of 3 different lots (20 tubes per lot) of 3.2% citrate blood tubes manufactured by Becton Dickinson, Greiner and Kima were studied. One additional lot from each manufacturer was assessed in triplicate (three series of 20 tubes), to assess within-lot variation. All tubes were first weighed empty and then filled with distilled water by a syringe, under ideal filling conditions. Filled tubes were weighed again, in duplicate. For each 20 tubes series, mean bias (deviation from the ideal tube filling volume) and imprecision (coefficient of variation; CV%) were calculated. All biases were within ±10%. Within-lot and between-lot variation in filling volume was acceptable, and comprised between 0.4 and 2.4%. Greiner tubes were the most accurate (bias, −1.0 to 2.4%), followed by Kima (bias, −7.8 to −5.9%) and Becton Dickinson (bias, −9.6 to 3.3%) tubes. The highest between-lot difference was noted for Becton Dickinson tubes (up to 12.9%), followed by Greiner and Kima tubes (up to 3.4 and 1.8%, respectively). Although coagulation tubes filling accuracy was within ±10% for all three tested manufacturers, the overall bias was found to be variable among manufacturers and lots. Major effort shall be made by blood tube manufacturers for improving standardization of their products.

Thrombin generation in different commercial sodium citrate blood tubes.

This study aimed to verify whether blood drawn into six different commercial coagulation tubes generated comparable results of thrombin generation. Blood was sequentially collected from 20 healthy subjects into different brand and draw volume 3.2% sodium citrate tubes (4.3 mL Sarstedt, 3.0 mL Greiner, 2.7 mL Becton Dickinson, 2.0 mL Kima, 1.8 mL Sarstedt and 1.0 mL Greiner). Thrombin generation was measured in plasma with the fully-automated ST Genesia analyzer using the weakest trigger (STG-BleedScreen). Different values of lag time (LT), time to reach thrombin peak (TP), thrombin peak height (PH) and endogenous thrombin potential (ETP) were commonly found in different tubes. Thrombin generation was the lowest in 4.3 mL Sarstedt tubes and the highest in 1.0 mL Greiner tubes. Other tubes displayed intermediate values. In multiple comparisons, LT was significantly different in 6/15 cases (40%), whilst PH, TP and ETP were significantly different in 14/15 (93%), 13/15 (87%) and 13/15 (87%) cases. The mean percent bias of LT, PH, TP and ETP ranged between -6% and +1%, -27% and +116%, -22% and +8%, and between -18% and +65%. The intra-assay imprecision of LT, PH, TP and ETP was exceeded in 0/15 (0%), 13/15 (87%), 6/15 (40%) and 13/15 (87%) comparisons. The correlation of LT, PH, TP and ETP values in different tubes ranged between 0.718-0.971, 0.570-0.966, 0.725-0.977 and 0.101-0.904. Blood collection for thrombin generation assays requires local standardization using identical tubes for brand and draw volume, and reference ranges calculated according to type of tubes.

Carotid Vibro-Compression: A Reperfusive Treatment for Acute Ischemic Stroke? Feasibility Data and a Potential Risk / Benefit Discussion in View to Animal Testing

Slow uncertain reperfusion in Acute Ischemic Stroke (AIS) by intravenous thrombolysis has prompted search for alternative therapies. Pressure/flow disturbances from rapid compressions (20 – 24 Hz) of a clotted flow tube remote from thrombosis site have shown to assist reflow. Carotid Vibro-Compression (VC), a potentially risky maneuver, may therefore enhance AIS reperfusion but verification is needed that vibro-compressions would transmit flow disturbances to the cerebral arteries. From a pool of 15 volunteers (mean age 48) carotid compressions (5 to 8 Hz, < = ~1 cm) (n =11) and vibration (30 Hz ~ 1 mm) (n = 10) were independently and together implemented with ipsilateral and contralateral Doppler flow monitoring of an intra-cranial artery. Doppler pulses from compressions were immediately demonstrated in all vessels (26 / 26) under all conditions. Pulses from vibration were observable in a majority of vessels (22 / 25), but signals were often challenging to obtain. Mean intra-cranial flow velocities trended to increase during compressions (46.1 ± 6 .4 cm/s vs. 44.0 ± 4.9 cm/s), but the difference was not statistically significant (p = 0.06). Carotid VC reliably transmits flow pulses to the cerebral vasculature, however vibration requires biofeedback to ensure device positioning. Carotid VC appears feasible for animal testing, but safety remains a major question.

Congenital Hypothyroidism in Neonates of a Tertiary Care Hospital.

Objective:To determine neonatal congenital hypothyroidism among neonates born in a tertiary care hospital of Lahore Pakistan.Methods:This cross-sectional study was carried out at Pathology Department of Allama Iqbal Medical College, Lahore in collaboration with Pediatrics and Gynecology & Obstetrics Department, Jinnah Hospital, Lahore Pakistan. A total of 770 babies were included in this study, both male and female. About 2 ml venous blood samples were collected aseptically from the neonates in sterile clotted tube. Serum was separated and serum TSH was determined by ELISA method.Results:Out of total 770 neonates, 48.9% were female and 51.0% were males with the ratio of 1:1.04. Neonatal congenital hypothyroidisim (TSH, >30 mIU/L), was observed in 0.4% (Frequency, 1:257) nenates, with the incidence rate of 1:257. Female to male ratio of hypothyroid neonates was 2:1. The mode of delivery vise distribution showed, among n=251 neonates born by normal delivery, only a single case of hypothyroidism was detected, and among n=519 neonates delivered by cesarean section, only two neonates were belong to hypothyroidism.Conclusion:The frequency of Congenital Hypothyroidism is notably higher in pediatric community than reported in most other countries. This result emphasizes the necessity of a nationwide screening program.

Analysis of amino acids in human blood using UHPLC-MS/MS: Potential interferences of storage time and vacutainer tube in pre-analytical procedure

ObjectivesTo investigate potential interferences of two pre-analytical variables, the storage time and the vacutainer tube, on the quantification of 20 amino acids using a UHPLC-MS/MS method. Design and methodsBlood samples from 25 apparently healthy subjects were collected into duplicate sets of EDTA-2K, EDTA-3K, coagulation, heparin and citrate tubes, and stored in capped vacutainer tubes at 4°C for 6h, 12h and 24h before sample analysis. A UHPLC-MS/MS method was established for simultaneous quantification of 20 amino acids. ANOVA for repeated measurement was conducted based on the model of Mauchly's test of Sphericity. Student's t-test was applied for comparison between amino acid concentrations obtained from different vacutainer tubes, and consistency of the results was checked through Bland-Altman difference plots and Passing-Bablok regression analysis. ResultsMost of the 20 amino acids showed a least concentration fluctuation with storage time in heparin plasma, followed by EDTA-3K and citrate plasma. The amino acid concentrations were significantly lower in citrate plasma and slightly higher in serum, compared with those in heparin plasma. No fixed bias was observed for amino acid concentrations in EDTA and heparin plasma, but the differences were mostly of statistical significance. Amino acid concentrations in EDTA-3K plasma achieved a good consistency with those in heparin plasma by UHPLC-MS/MS analysis. ConclusionsStorage time and vacutainer tube were important variables for amino acid analysis. They should draw researchers' attention and then be controlled in good laboratory practice to reduce pre-analytical errors.