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Coordinator's report: an assessment of the functional activity of human Rh monoclonal antibodies after their evaluation by nine laboratories.

The functional activity of 55 anti-D and 26 MAbs of other Rh specificities was determined as part of the Third International Workshop and Symposium on Monoclonal Antibodies Against Human Red Blood Cell and Related Antigens. Most MAbs were IgG1 (45 anti-D, 16 Rh). There was a large range of anti-D and IgG concentrations of the anti-D MAbs (0.4-1680 IU/ml, 1.5-400 micrograms/ml). Correlation between quantitative data from different laboratories was good. Six laboratories tested the anti-Ds in monocyte ADCC assays. Methods varied greatly, especially the effector cells used, the methods of red cell sensitisation, the effector to target cell ratio and the assay incubation times. There was some correlation of results between most laboratories, but results were better when similar assays were performed. The extent of monocyte-mediated haemolysis was related to the number of molecules of IgG anti-D bound to the red cells. Monocyte phagocytosis assays resulted in a greater variability in results between the four evaluating laboratories, though IgG3 MAbs were found more active than IgG1, and mostly promoted red cell adherence rather than phagocytosis. Two monocyte chemiluminescence assays found comparatively low activity of the IgG3 MAbs; the most active IgG1 anti-Ds were MAbs 93 and 95. Correlation between different monocyte-mediated assays was generally poor unless the assays were performed by the same laboratory. Results of a macrophage ADCC assay showed that the IgG3 anti-D's promoted greater haemolysis than most IgG1 MAbs. Only two IgG1 anti-D MAbs (70 and 104) mediated high haemolysis in this assay. Lymphocyte ADCC assays were utilised in four laboratories. The IgG3 antibodies exhibited low activity but one IgG1 anti-D (104) consistently mediated high haemolysis. There was no relationship between quantitation and haemolysis in this assay. A few MAbs exhibited low or no activity in most assays, mainly due to low quantitation. Most of the MAbs of non-D specificity mediated low functional activity which in most cases was not due to low quantitation. However two MAbs, 24 (anti-CcEe) and 25 (anti-CD), consistently showed good activity. This study highlighted the great variability in assay methodology between the nine participating laboratories, which resulted in some apparent differences in functional activity of some of the MAbs. The use of standardised methods as well as fewer MAbs tested and quantitated at different concentrations may help to determine the relevant factors contributing to IgG function.

Antibodies, Monoclonal↗

Working with male rodents may increase risk of allergy to laboratory animals.

BACKGROUND: Our aim was to study the risk of laboratory animal allergy (LAA) among research staff working in laboratories separate from the animal confinement area. The roles of atopy and exposure intensity in LAA were studied with special regard to exposure to male rodents, who excrete higher levels of urinary allergens than female rodents. METHODS: Eighty rodent-exposed subjects gave blood samples for the analysis of total IgE, Phadiatop, and specific IgE against rat (RUA) and mouse urinary allergens (MUA), and answered questionnaires. Air samples were collected for RUA and MUA aeroallergen measurement in both laboratories and animal confinement facilities. RESULTS: Twenty percent of the subjects had IgE >0.35 kU/l to RUA and/or MUA, and 32% had experienced animal work-related symptoms, although 90% of aeroallergen samples from the research department laboratories were below the detection limit (<0.26 ng RUA per m(3) and <0.8 ng MUA per m(3)). Atopy (positive Phadiatop), total IgE >100 kU/l, other allergies (especially to other animals), or more than 4 years of exposure significantly increased laboratory animal sensitization and symptoms. Working with mainly male rodents gave odds ratios (95% CI) of 3.8 (0.97-15) for sensitization and 4.4 (1.4-14) for symptoms. Subjects with both exposure to mainly male rodents and atopy or elevated total IgE had a 10-fold higher frequency of sensitization than exposed subjects with neither risk factor. CONCLUSION: A majority of subjects with a combination of exposure to mainly male rodents and atopy or elevated total IgE developed sensitization to and symptoms from laboratory animals. Current low exposure seems to maintain the presence of specific IgE. Further measures must be undertaken to provide a safe workplace for laboratory animal workers.

Adult↗

Boston City Hospital and the Thorndike Memorial Laboratory: the birth of modern haematology.

Established in 1923, the Thorndike Memorial Laboratory at Boston City Hospital was the first clinical research laboratory in a municipal hospital in the United States of America. Minot and Castle, who were the second and third directors of the Laboratory, were pioneer haematologists and clinical investigators of the highest calibre who created an atmosphere at the Laboratory that would foster patient-centred research and attract the best physician-scientists to work and train there. The haematology research division of the Laboratory made important original contributions to the understanding of the pathophysiology of anaemia, the mechanisms of red cell and platelet destruction and the phagocytic role of the spleen, the nature of haemoglobin (normal and sickle cell), the nature of haemophilia and its therapy and the early classification of lymphoma. It contributed to the Thorndike Memorial Laboratory's worldwide reputation as a model research laboratory and established its reputation as the birthplace of modern haematology.

Biomedical Research↗

Effects of chlorpyrifos on field- and laboratory-developed estuarine benthic communities.

Macrobenthic animal communities, developed in sand-filled aquaria in the laboratory and in the field, were exposed to various concentrations of the insecticide chlorpyrifos, and effects on community structure were assessed. Laboratory communities were continuously exposed to the toxicant for 8 wk during colonization by planktonic larvae in unfiltered Santa Rosa Sound, Fla., seawater. Field communities were developed for 8 wk in aquaria placed in Santa Rosa Sound, then removed to the laboratory for exposure to chlorpyrifos for one week. Abundance of arthropods was significantly diminished (alpha = 0.05) by measured concentration of chlorpyrifos greater than or equal to 0.1 microgram/l in water in laboratory communities and by 5.9 microgram/l in water in field communities. Numbers of annelids and chordates in contaminated aquaria were not reduced by the highest concentrations of chlorpyrifos tested, 8.5 microgram/l in laboratory-colonized aquaria and 5.9 microgram/l in field-colonized aquaria. One species of annelid, Cistenides gouldii, was more abundant in field aquaria receiving 1.0 microgram/l or 5.9 microgram/l than in the control and lowest concentration. Molluscan larvae colonizing laboratory aquaria were sensitive to greater than or equal to 0.1 microgram/l; however, later developmental stages characterizing field aquaria were not sensitive to less than or equal to 5.9 microgram/l. Although only 20 of 78 animal species appeared in both laboratory and field communities, sensitivity of animals in these tests and in single species tests could be compared.

Animals↗

Susceptibility of laboratory and field strains of four stored-product insect species to spinosad.

Two field strains of the Indianmeal moth, Plodia interpunctella (Hübner); red flour beetle, Tribolium castaneum (Herbst); and lesser grain borer, Rhyzopertha dominica (F.), and one field strain of the rusty grain beetle, Cryptolestes ferrugineus (Stephens), were collected from hard red winter wheat stored on farms in northeastern Kansas. Fifty eggs of P. interpunctella and 25 beetle adults of each species were exposed to 100 g of untreated wheat or wheat treated with various rates of spinosad, to determine susceptibility of the field and corresponding insecticide-susceptible laboratory strains. Mortality of beetle adults and P. interpunctella larvae was assessed after 7 and 21 d postinfestation, respectively. Field strains of P. interpunctella, C. ferrugineus, and T. castaneum were less susceptible to spinosad than the corresponding laboratory strains. The LD50 and LD95 values for P. interpunctella and C. ferrugineus field strains were 1.7-2.5 times greater than values for corresponding laboratory strains. Adults of both laboratory and field strains of T. castaneum were tolerant to spinosad, resulting in <88% mortality at 8 mg/kg. The LD50 and LD95 values for the field strains of T. castaneum were 2.0-7.5 times greater compared with similar values for the laboratory strain. The field and laboratory strains of R. dominica were highly susceptible to spinosad, and one of the field strains was relatively less susceptible to spinosad than the laboratory strain. Our results confirm a range of biological variability in field populations, which is consistent with findings for other compounds, and underscores the need to adopt resistance management programs with stored grain insect pests. The baseline data generated on the susceptibility of the four insect species to spinosad will be useful for monitoring resistance development and for setting field rates.

Animals↗

Inter-laboratory test program (PTI) in 'neutron dosimetry'.

The Laboratory of External Dosimetry Studies and Researches (LRDE), associated laboratory to the Bureau National de Métrologie, is strongly involved in the reference establishment and calibration of instruments in neutron radiation fields provided by 241Am-Be, 252Cf and (252Cf + D2O)/Cd sources. In 1997, on the request of COFRAC, the LRDE has managed an Inter-Laboratory Test Program in 'neutron dosimetry' to compare the different standard calibration methods. The transfer instrument used was a "Harwell Monitor N91". The results of the participant were in good agreement but the associated uncertainties need to be revised by some of laboratories. According to the Mutual Recognition Arrangement signed in 1999, laboratories for metrology will have to assure the traceability of their references. In 2003-2005, the LRDE organises, with National Physical Laboratory (UK) and Physikalisch-Techische Bundesanstalt (Germany), an international comparison in 'neutron dosimetry'. Two transfer instruments, a "2202D" rate metre and a "Harwell Monitor N91", will be sent in turn to the participant laboratories for measurements at their own neutron sources facilities.

Body Burden↗

Minimizing admission laboratory testing in trauma patients: use of a microanalyzer.

OBJECTIVE: Routine admission laboratory test protocols in injured patients are costly and involve excessive phlebotomy and turnaround time. The purpose of this prospective study was to evaluate the utility of (1) a microanalyzer, NOVA-SP5 (which provides rapid results on minimal blood volume), and (2) each component of our standard laboratory test battery. METHODS: Laboratory test results for 200 consecutive injured patients admitted to a level I trauma center were evaluated by paired sample analysis. Our standard battery [60 mL: ($348): type and screen, complete blood count, PT/PTT, electrolytes, BUN, creatinine, glucose, calcium, amylase, ethanol level, and arterial blood gas] run "stat" in the central laboratory was compared to the microanalyzer profile [< 1 mL: ($182): hemoglobin, hematocrit, electrolytes, glucose, Ca2+, and arterial blood gas] run by the trauma team in the resuscitation area. Patient data and laboratory turnaround time (from time of admission to time results obtained) were recorded. Data were analyzed by linear regression. RESULTS: Components of the paired samples correlated well (r2 0.78 to 0.99). Turnaround times were 64 (+/- 3) and 6 (+/- 1) minutes for standard analysis and microanalysis, respectively. Only two of the 26 patients requiring emergent surgical procedures had standard results available preoperatively. These patients had twice as many laboratory abnormalities as the remainder. Minimal diagnosis or intervention resulted from those values exclusive to standard analysis (white blood count, amylase, ethanol level, BUN, creatinine, platelet count, PT, and PTT). Six of ten abnormal BUN or creatinine results normalized, including two values in patients who received contrast for portable intravenous pyelography, and in all patients without a history of hypertension or diabetes. Platelet count and PT/PTT were normal in 85% of non-head-injured patients, compared with 58% of those with GCS score < or = 8. CONCLUSIONS: Microanalysis is accurate, expedient, conserves blood, and is sufficient for evaluation of most trauma patients. Those with hypertension, diabetes, or severe head trauma may require additional testing. Routine use of this technique could reduce cost substantially ($16,000/100 patients). The role of microanalysis in follow-up laboratory evaluation of injured patients remains to be elucidated.

Adult↗

Ambulatory blood pressure monitoring and laboratory reactivity in type A behavior and components.

Type A coronary-prone behavior has been characterized previously by cardiovascular hyperreactivity to laboratory stressors. The objectives of this study were: 1) to determine whether cardiovascular patterns under more naturalistic circumstances in the field were altered in Type A subjects, and 2) to determine whether these field patterns paralleled cardiovascular patterns to a series of stressors in the laboratory. Thirty-three healthy Type A and B men underwent 24-hr ambulatory blood pressure and heart rate monitoring during a normal day of activities. These subjects were also tested in the laboratory for blood pressure and heart rate responses to a series of stressor tasks: 1) cognitive (mental arithmetic), 2) perceptual motor (reaction time), and 3) physical (handgrip). Type A behavior and stylistic components were rated by structured interview. All studies were conducted double blindly. Type A subjects demonstrated greater cardiovascular reactivity to the laboratory cognitive stressor, but there was a mixture of differences between As and Bs in variability of ambulatory blood pressure and heart rate during different periods of the day. Yet, there were no differences in average blood pressure levels. The components of verbal competition and loudness of voice were positively associated with elevated average blood pressure and variability in the field, respectively, but not with laboratory hyperreactivity. Thus, this study confirmed laboratory-induced cardiovascular hyperreactivity in Type A behavior but, within its limitations, did not find a similarity between laboratory and field cardiovascular response patterns in these behavioral groups. The ambulatory blood pressure monitoring findings in the component groups may have prognostic significance and deserve further study.

Adult↗

An interprovincial external quality assessment of the ability of Canadian laboratories to detect the vancomycin and penicillin resistance of Enterococcus faecium D366.

OBJECTIVE: To evaluate the ability of Canadian laboratories to identify enterococci and detect low-level resistance to penicillin, ampicillin and vancomycin in five provinces and two territories by two external quality assessment schemes. METHODS: Enterococcus faecium, strain D366, with minimum inhibitory concentrations for vancomycin and penicillin of 32 and 16 mg/L respectively, was distributed during a routine proficiency survey. Laboratories were required to culture and identify the isolate and to test antimicrobial susceptibility. Participants were assessed against consensus reference values. RESULTS: Three hundred and sixty-four hospital, commercial and public-health laboratories participated, using their established procedures for patient samples. The isolate was identified to the species level by 222 (61%) laboratories and to the genus level by a further 98 participants. Forty-four failed to meet the expected standard. Vancomycin resistance was detected by 94%. Those reporting a falsely susceptible result used disk diffusion testing. Penicillin resistance was noted by 250 of 258 laboratories reporting on this agent. An incorrect ampicillin-susceptible finding was reported by 62 of 147 laboratories using automated microdilution or agar dilution methods. CONCLUSIONS: Most laboratories identified the isolate to an appropriate level. Detection of low-level vancomycin and penicillin resistance was achieved by the majority. Ampicillin resistance was less readily detected.

Journal Article↗

Global surveillance and the value of information: the case of the global polio laboratory network.

Effective control and eradication of diseases requires reliable information from surveillance activities, including laboratories, which typically incur real financial costs. This article presents data from a survey we conducted to estimate the costs of the Global Polio Laboratory Network (GPLN), which currently supports aggressive global surveillance for acute flaccid paralysis (AFP) to detect circulating polioviruses. The Global Polio Eradication Initiative (GPEI) of the World Health Organization (WHO) provides resources for some of the laboratory network costs, but the total cost of the network remains relatively poorly characterized given the limited documentation of national contributions. We surveyed network laboratories to quantify AFP surveillance support costs and provide data for cost estimates of potential posteradication surveillance policies related to the laboratories. We estimate that the GPLN currently requires millions (US dollars 2002) in total support annually, and that half of the support for national and regional reference laboratories comes from external donors through the WHO or bilateral agreements and half from within nations that host those laboratories. The article also presents the framework for considering the value of information from this global surveillance network and suggests that the expected value of surveillance information from the GPLN currently exceeds its costs. We also provided important insights about how the value of information may change after successful eradication of wild polioviruses.

Global Health↗

Inter and intra laboratory variation of digoxin radioimmunoassay in Sweden.

Samples from two pools were sent 10 times to 27 laboratories for assay of digoxin. One pool contained digoxin 2.60 nmol/l in normal plasma (SP); the other was pooled plasma from patients treated with digoxin (PP). Ten radioimmunoassay (RIA) methods were used. The mean of SP assays was 2.59 nmol/l, not significantly different from 2.60 nmol/l. The mean of PP determinations was 2.46 nmol/l. Within each of the 10 assay rounds, the concentrations showed an almost twofold variation and S.D. averaged 0.33 nmol/l and 0.31 nmol/l for SP and PP respectively. Significant differences (P less than 0.001) were found between mean concentrations obtained for the pools at various laboratories (SP range 2.15-2.85 nmol/l; PP range 2.12-2.72 nmol/l). The laboratory means obtained for SP and PP correlated significantly (P less than 0.001). Nevertheless, significant (P less than 0.01) variations between laboratories were found also concerning the mean difference between SP and PP concentrations. The interassay SD of the assays differed significantly between laboratories (range 0.05-0.61 nmol/l. Between and within groups of laboratories using the same RIA method and between various types of laboratories, differences were also found concerning both accuracy and precision of the assays. It is concluded that a better control of digoxin assay is needed.

Digoxin↗

In vivo persistent pigment darkening method: a demonstration of the reproducibility of the UVA protection factors results at several testing laboratories.

BACKGROUND/PURPOSE: The aims of the present studies were to check that persistent pigment darkening (PPD) method can produce accurate and reproducible results for high UVA protection factors (UVAPF) and to provide data on the variability between laboratories and on the influence of skin types. METHODS: The Japanese Cosmetic Industry Association (JCIA) PPD method was used to determine the UVAPF in different laboratories of different sunscreen formulations with increasing UVAPF. Two formulations were tested at seven independent laboratories and five products within two laboratories. The influence of skin types on the UVAPF of two products was tested within one laboratory on two panels of volunteers. All laboratories complied with the JCIA method specifications. RESULTS: Reproducible results have been obtained between the different labs and a low and satisfactory variability was achieved with a panel size of 10 subjects. Furthermore, skin type was demonstrated to have no influence within the defined selection criteria. CONCLUSIONS: From this multiple center testing, the PPD method has been shown to be appropriate for testing sunscreen formulations with UVAPFs above 20. It is reasonable to expect that test results will be consistent if an identical protocol is used between laboratories.

Observer Variation↗

Evidence that the National Committee for Clinical Laboratory Standards disk test is less sensitive than the screen plate for detection of low-expression-class methicillin-resistant Staphylococcus aureus.

A low-expression-class methicillin-resistant Staphylococcus aureus strain (strain 9302-2) was sent to 207 laboratories as part of the bacteriology component of the Laboratory Proficiency Testing Program of Ontario. An incorrect (susceptible) result was reported by 16 of 76 (21%) of laboratories that used the National Committee for Clinical Laboratory standards disk test, whereas 1 of 104 laboratories that used other methods reported an incorrect result (P < 0.05). Experiments showed discrepancies in the disk test results given by Mueller-Hinton agars from three manufacturers. We advise that laboratories should use a low expression-class methicillin-resistant S. aureus isolate as a control for the National Committee for Clinical Laboratory Standards disk test.

Humans↗

Quality assessment of blood glucose monitors in use outside the hospital laboratory.

The quality of analytical results required in clinical practice is generally dictated by analytical rather than clinical criteria. The increasing availability of analytical technology that can be used outside the hospital laboratory challenges this situation. In an attempt to highlight this dilemma one method of glucose analysis outside the laboratory has been assessed using a pilot external quality assessment scheme (EQAS) and the results have been compared with the performance of the same machines in the laboratory. In the laboratory reliable results were obtained when the Glucometer was used by experienced laboratory personnel giving a good correlation of results (r = 0.96) when compared with an automated method. The results of the EQAS revealed that 44% of hospital ward glucose estimations and 63% of general practice estimations would be considered unsatisfactory by conventional laboratory criteria--that is, greater than +/- 2 SD of the mean laboratory result. This level of performance must, however, be considered in relation to the number of times that a clinically misleading result is obtained. No analytical system should be used in clinical practice without a continuous, objective assessment of its performance.

Blood Glucose↗

Laboratory contamination of Mycobacterium tuberculosis cultures.

The purpose of this study was to investigate possible laboratory contamination of Mycobacterium tuberculosis cultures which resulted in the misdiagnosis of tuberculosis. We have investigated three cases in which a patient's culture was positive for M. tuberculosis but there was not a high clinical suspicion for disease. In each instance, another patient with clinically obvious pulmonary tuberculosis had specimens cultured concurrently within the same clinical laboratory. The isolates from both the obvious cases of tuberculosis and the suspect cases were obtained through the State of Alabama TB Laboratory, but these isolates originated at a commercial laboratory, a community hospital laboratory, and at a university hospital. MTB isolates were fingerprinted by probing for the insertion sequence IS6110. With each of the three pairs of isolates (case and suspicious case), identical IS6110 banding patterns were found suggesting identical MTB strains. Because the patients were geographically separated, it is strongly suspected that laboratory contamination of M. tuberculosis cultures resulted in the three suspect cases being diagnosed with tuberculosis. These findings indicate that positive M. tuberculosis cultures resulting from laboratory contamination can occur.

Adult↗

The safety of performing diagnostic cardiac catheterizations in a mobile catheterization laboratory at primary care hospitals.

The benefits of mobile cardiac catheterization laboratories include keeping patients closer to their families, communities, local hospitals, and primary care physicians while receiving services comparable to those available at tertiary centers. However, there are very few studies regarding the safety of performing cardiac catheterization in mobile laboratories at remote locations. Thus, the authors performed an observational study of 1,775 consecutive patients undergoing a diagnostic cardiac catheterization in a mobile catheterization laboratory at primary care hospitals served by the Appleton Heart Institute (AHI) from August 1, 1991, to December 31, 1998. Twenty-three percent (1,775/7,637) of all AHI diagnostic cases in this time period were performed in the mobile catheterization laboratory. Urgent transfer to the tertiary care facility via ambulance or helicopter was used for 2.3% of patients (n = 41). The overall complication rate was 1.2% (n = 21). Of the patients who underwent cardiac catheterization in the mobile laboratory, 32.6% (n = 579) were subsequently referred for interventional or surgical revascularization. There were no deaths. Cardiac catheterizations can be performed safely in a mobile laboratory at primary care hospitals, provided that immediate transfer is available for those in need of urgent intervention or revascularization and that unstable patients are not studied in the mobile laboratory.

Adult↗

Assessment of the practicability and analytical performance of a point-of-care affinity chromatography haemoglobin A1c analyser for use in the non-laboratory setting.

BACKGROUND: Haemoglobin A(1c) (HbA(1c)) is a pivotal pathology test used around the world for the long-term management of patients with diabetes. Point-of-care testing (POCT) provides a convenient means for conducting HbA(1c) testing outside the laboratory. METHODS: The practicability and analytical performance of the Micromat II POCT HbA(1c) analyser (Bio-Rad Laboratories, USA), which has affinity chromatography as its methods principle, was evaluated in Australia and compared with the DCA 2000 POCT device (Bayer Australia) and a laboratory-based high-performance liquid chromatography (HPLC) method. RESULTS: Overall between-day imprecision over 10 days was 1.9% for the laboratory HPLC method, 2.2% for the DCA 2000 and 7.0% for the Micromat II. In a second study over the same time period, the Micromat II's imprecision was 6.4%. The mean difference between the Micromat II and the laboratory method in a patient comparison (n = 100) was -0.25% (lower and upper limits of agreement -1.79 to 1.30). CONCLUSIONS: The imprecision obtained with the Micromat II was inferior to both the DCA 2000 and laboratory methods and did not meet current internationally accepted precision goals for this analyte. The Micromat II's poor imprecision can be explained by the high degree of technical expertise needed to perform the test; its use by non-laboratory health professionals such as nurses and Aboriginal health workers in rural and remote Australia cannot be recommended.

Australia↗

Accuracy of the avosure PT pro system compared with a hospital laboratory standard.

OBJECTIVE: To compare international normalized ratio (INR) values obtained using the AvoSure PT Pro point-of-care (POC) system with those obtained using a standard laboratory method. METHODS: Forty-one INR values obtained from the POC system were compared with those obtained from a standard laboratory method. The POC method was evaluated for both laboratory and clinical agreement. To evaluate laboratory agreement, various analyses were used, including mean-squared prediction error (MSE) and mean prediction error (ME), Bland-Altman analysis, correlation, and paired t-test comparing group INR means. For clinical accuracy, discrepant pairs were identified and evaluated to determine whether dosage adjustments would have been needed based on values obtained. RESULTS: The POC system demonstrated modest precision (MSE = 0.147, 95% CI 0.065 to 0.228) and relatively little bias (ME = 0.090, 95% CI -0.025 to 0.205). Bland-Altman analysis also suggested good agreement at average INRs from 2.0 to 3.0. At average INR values >3.0, the POC system consistently overestimated INR. Values obtained with the POC system were significantly correlated with those obtained from the hospital laboratory (r = 0.77; p < 0.001). Similarly, mean +/- SD POC INR did not differ significantly from the laboratory-determined INR (2.45+/-0.59 vs. 2.37+/-0.48, respectively; p = 0.176). Regarding clinical accuracy, the values clinically agreed in 85.4% of the cases. CONCLUSIONS: The AvoSure PT Pro POC system appears to be useful for INR values within the 2.0-3.0 range, but values outside of this range should probably be confirmed with a standard laboratory method.

Anticoagulants↗