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[Comparative study of digoxin determination by E.L.I.S.A. and radioimmunoassay techniques (author's transl)].

This study is based on a double blind interlaboratory comparison between radioimmunoassay and E.L.I.S.A. digoxin determination. The correlation between digoxin values found with these two methods is good (r = 0.96 for therapeutic and toxic ranges 1,0 ng/ml - 5,5 ng/ml). The results indicate good repeatability, reproducibility and precision. The determination by E.L.I.S.A. can be performed with sera or plasma. The presence of haemolysis makes no appreciable difference in results. No variation is observed when different kits are used from an identical lot. However digoxin gives an important cross reactivity with digitoxin in Enzymeimmunoassay. Therefore it is necessary to know exactly the digitalis glycoside before the determination in order to avoid significant error.

Digoxin

[Serum amylase determination : automated continuous flow method (author's transl)].

A simple automated assay was described for the determination of serum amylase using a chromogenic substrate. The reaction was stopped by filtration across a cellulose nitrate membrane. The analytic parameters of this method were studied. Repeatability, reproducibility and precision were convenient. The calibration curve was linear up to 850 U/I. An excellent correlation was obtained after comparison with the manual method (Phadebas amylase test). The effects of haemolysis and very high ascorbic concentrations of bilirubin, glucose, uric acid and ascorbic acid were negligible on the value measured. The serum turbidity was alone to induce a variation by excess.

Amylases

[Susceptibility of Ornithodoros papillipes (Argasidae) ticks to the rickettsia Coxiella burnetii].

A definition of susceptibility of an arthropod to an infectious or invasional agent is given. The susceptibility means a capacity of the arthropod for infection in a result of which the agent either completes a part of its life cycle in the organism of its invertebrate host or reproduces repeatedly in the organs and tissues of its internal medium causing or not causing a noticeable damage. On the model of the tick O. papillipes and Q. fever agent there were shown differences in the resistance of arthropods to Rickettsiae depending on the developmental phase of the invertebrate host. Under otherwise equal conditions the susceptibility of nymphs of O. papillipes to C. burnetii was in nymph II 146 times, in nymph I 280 times and in larvae 50 000 times less than in nymph IV. It has been established that nymphs I, II, and IV engorge blood only 2.5, 14 and 50 times more respectively than larvae. It shows that the success of infection depends to a larger extent on the degree of susceptibility of an arthropod to a corresponding agent than on the quantity of engorged infected blood. The formula of estimation of agent concentration necessary for successful infection of 50 per cent of ticks is given.

Animals

[A rapid technique for extraction of urinary 17-kéto and 17-hydroxy-corticosteroids by chromatography (author's transl)].

The extraction ot total urinary 17-keto and 17-hydroxy steroids has been effected by a rapid procedure involving pre-packed kieselguhr column chromatography. This work successively studies the correlation of this technique with the classical extraction by mechanical agitation (42 and 61 subjects), its reproducibility, repeatability and recovery of added internal standards. The proposed method seems convenient for routine clinical analysis.

17-Hydroxycorticosteroids

Phleborheography--results of a ten-year experience.

Phleborheography (PRG) is a physiologic volumetric plethysmographic technique that was developed for the diagnosis of lower extremity deep venous thrombosis. During the past 10 years, 21, 626 extremities have been studied with 709 corresponding venograms. Overall data reveal a sensitivity of 92% (247/268) and specificity of 95% (418/441). There were 21 false negative PRG results, and 12 of these were due to isolated calf vein thrombosis. There were 23 false positive results, and 11 of these were considered true errors of the technique. When isolated calf vein thrombosis was disregarded, the sensitivity increased to 95%. PRG detected 45 of 57 (79%) isolated calf vein thrombi. The modified criteria for interpretation included respiratory waves and baseline elevation as the major criteria and prominent arterial pulsation and foot emptying as the minor criteria. PRG has recently been used to monitor the duration of thrombolytic therapy. While data are limited, this appears to be a promising contribution to the quantification of individual response to lytic therapy and may indicate an objective end point for treatment. PRG is a sensitive and specific method of detecting deep vein obstruction. It is a painless, reproducible, repeatable technique that requires minimal patient cooperation. PRG is a versatile laboratory technique that can be used for a variety of applications in addition to the diagnosis of deep vein thrombosis.

False Negative Reactions

[Assay of 3-methoxy 4-hydroxyphenyl ethylene glycol (MHPG) and its conjugates in the urine (author's transl)].

Urinary MHPG was assayed by gas chromatography using a flame ionisation detector. The technic proposed was reproducible, repeatable and linear. The frequent values determined in adult subjects of both sexes were 8.64 1.34 mumol/24 h without any difference between the sexes. On the other hand, urinary excretion of total MHPG showed a positive correlation with age. It was excreted in the conjugated form, of which 50 p. cent was sulphoconjugated, 41 p. cent glucuroconjugated, and 9 p. cent free. Owing to its simplicity, this method may be applied to the study of mood disorders.

Adult

[Methods for determining lead and cadmium in blood; cadmium, copper, nickel and chromium in urine using flameless atomic absorption spectrometry].

The estimation of environmental and occupational exposure to metals is based on the determination of metal concentrations in biological material. This paper describes methods for determining lead and cadmium in blood, and cadmium, copper, nickel and chromium in urine. The methods are evaluated in view of their linearity, precision, repeatability, reproducibility and detectability. The reliability of the methods was verified by determining lyphilized standards of known metal concentrations (Behring). The reproducibility expressed by the value of relative standard deviation ranged from 1.2% to 7.8%. In the case of lead and cadmium determinations, the reliability of results was confirmed due to the participation of our laboratory in the UK NEQAS external control project, the Queen Elizabeth Hospital, Birmingham. The results of external quality control (MR VIS = 19 for lead, as compared with the mean = 41 for all the project participants and MR VIS = 29 for cadmium, mean = 62) prove undoubtedly correctness of the methods applied. A small volume of sample to be analyzed is their major advantage, and due to adopted analytical parameters the methods can be used to determine metals in biological material in order to assess occupational or environmental exposure.

Cadmium

TFIIIA and DNA supercoiling: toward resolving a controversy.

DNA supercoiling in eukaryotes is mediated by the process of chromatin assembly. This process couples the binding and wrapping of DNA around nucleosomal core particles and the enzymatic activity of DNA topoisomerase(s). Kmiec and Worcel (1985) reported that the binding of an RNA polymerase III transcription factor (TFIIIA) could accelerate DNA supercoiling catalysed by a Xenopus laevis cell-free extract. Although the reaction was repeatedly reproduced in the parent laboratory by numerous workers, another laboratory, interested in the molecular biology of TFIIIA, failed to reproduce TFIIIA-mediated supercoiling or gyration. In this review, an attempt is made to transcend personal beliefs and describe the experimental approaches used to clarify this issue. The original variability lay in the amount of endogenous RNA within the cell free extract and the concentrations of MgCl2 used to conduct the experiments.

Adenosine Triphosphate

French multicentric evaluation of recombinant tissue factor (recombiplastin) for determination of prothrombin time.

Recombiplastin, a recombinant a human tissue factor, elaborated by Ortho Diagnostic Systems, produced by Baculovirus and relipidated with highly purified phospholipids, was tested as a new reagent for determining prothrombin time (PT) in a French multicentric study. Its intralaboratory performances, including sensitivity, repeatability, reproducibility and stability, were explored to establish whether its use would reduce the interlaboratory dispersion of PT values, and therefore improve the standardization of oral anticoagulant treatment. The 9 university hospital hematology laboratories involved in this study used the same type of instrument (KC 10). For 10 consecutive days, they determined PTS on a normal plasma pool, plasma dilutions of 1/2, 1/3 and 1/8, 3 identical lyophilized calibrated plasmas, as well as plasmas from 20 normal subjects, 50 patients on oral anticoagulant therapy with Recombiplastin which has an International Sensitivity Index (ISI) of 1, and 2 commercial thromboplastin extracts (ISI #1 or 2). In the patients on anticoagulants, factors VII, X and V were measured when results were conflicting. The intra and interlaboratory reproducibilities of Recombiplastin, calculated on the basis of either PTS expressed in seconds, or of the International Normalized Ratio (INR), were good, with coefficients of variation (CV) similar to those observed with the 5 other reagents used by the different laboratories (2% < CV < 8%). The stability of Recombiplastin was excellent, with no variation in PT after 72 h of incubation at 37 degrees C. A normal PT of 12 s was obtained with Recombiplastin, similar to the values found for the reagents with ISI #2.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral

[The reticuloendothelial system. An overview of function, pathology and recent methods of measurement].

The Reticuloendothelial System (RES) consists of cells descending from the monocytes which are able to perform phagocytosis of foreign materials and particles. 90% of the RES are located in the liver. The most important function of the RES ist phagocytosis, but it also takes part in cytotoxicity against tumor cells and has a function in the regulation of the immune system. The importance of the RES is obvious in diseases of the bile ducts. An obstruction can lead to a decrease in the function of the RES. Beside a mathematical model there are also a few experimental methods to measure the phagocytic capacity and thus the efficiency of the RES. They are based on a clearance study of the amount of particles absorbed by the RES. This study can e.g. be done with Nanocoll which has been marked with 99mTc. But instead of the bloody clearance, a scintigraphic study using a gamma camera can be performed. With that, the increase time, the liver extraction fraction and the steepness of increase can be determined. Such a test could be of clinical relevance as a parameter for the prognosis of patients with trauma, sepsis or shock. The function of the RES in the pathogenesis of the biliary pancreatitis could also be examined closer with the help of such a test. The requirements a test of the RES should meet are reproducibility, repeatability and practicability.

Animals

[Morphologic changes in the chorio-allantoic membrane and organs of chick embryos inoculated with blood of patients with systemic lupus erythematosus (concerning the viral etiology of systemic lupus erythematosus)].

Morphological studies of 14 chorionallantoic membranes and organs of chick embryos which had been infected on the 7--8th day of incubation with leucocytic mass from patients with systemic lupus erythematosus were carried out. Corresponding virological and morphological control studies were performed. The changes observed were repeatedly reproduced in multiple inoculations (from the 2nd to the 16th passage). Apart from morphological changes, which may be listed among general pathological processes (impairment of the circulation, dystrophy, proliferation), changes testifying to a possible cytopathic effect disintegration of nuclei, formation of gigantic cells and multinuclear symplasts were also noted. The most pronounced changes, both in the chorionallantoic membrane and in internal organs, were observed on the 3rd--5th passage and on the 3rd--5th day of incubation following inoculation. They correlated with the clinical activity of the process in patients from whom the blood for inoculation had been taken. The data obtained justify the assumption concerning the existence in the blood of the patients with systemic lupus erythematosus a transplantable agent producing a cytopathic action, and may be considered as a new indirect corroborative evidence in favour of the concept of the viral nature of systemic lupus erythematosus.

Animals

Substrate supporting disc method for confirmed detection of total coliforms and E. coli in all foods: collaborative study.

The ColiComplete substrate supporting disc (SSD) method for simultaneous confirmed total coliform count and Escherichia coli determination in all foods was compared with AOAC most probable number (MPN) methods, 966.23 and 966.24. Twenty-nine laboratories participated in this collaborative study in which 6 food types were analyzed. Four food types, raw ground beef, pork sausage, raw liquid milk, and nut meats, were naturally contaminated with coliform bacteria. Two foods, dry egg and fresh frozen vegetables, were seeded with coliforms. Three food types, ground beef, raw liquid milk, and pork sausage, were naturally contaminated with E. coli. Although pork sausage was naturally contaminated, the level was very low (<10/50 g); therefore, additional E. coli were inoculated into 1 lot of this food type. Three food types, nut meats, dry egg, and fresh frozen vegetables, were inoculated with E. coli. For naturally contaminated samples, duplicate determinations were made on 3 separate lots for each food type. For inoculated samples, low, medium, and high contamination levels plus uninoculated control samples were examined in duplicate. Data were analyzed separately for total coliform bacteria and for E. coli. Mean log MPN counts were determined by the SSD method and the appropriate AOAC MPN method. Results were then analyzed for repeatability, reproducibility, and mean log MPN statistical equivalence. Results were statistically equivalent for all total coliform levels in all food types except frozen vegetable and raw nut meat uninoculated control samples and 1 lot of pork sausage where the SSD method produced statistically significant greater numbers. For the E. coli determinations, results were statistically equivalent across all samples and all levels for each food type. The SSD method has been adopted first action by AOAC International for confirmed detection of total coliforms and E. coli in all foods.

Animals

[Adaptation of the measurement of plasma and erythrocyte glutathione peroxidase activity on Hitachi 911].

The aim was to automate the assay of plasma and erythrocyte glutathione peroxidase (GSH-Px) activity using the Hitachi 911 (Boehringer Mannheim). There is a need for a reliable and rapid technique for the determination of GSH-Px activity to evaluate 'the oxidative stress'. The technique used was that proposed by the Randox laboratories (Randox Laboratories Ltd, Ransel test, reference: RS-505). The Hitachi 911 was programmed by adapting the manual Randox technique. Repeatability, reproducibility and linearity were satisfactory, and usual values for plasma and erythrocyte GSH-Px activity were determined. Automation of the Ransel test for determination of GSH-Px activity was very useful for serial assays. In addition, the technique is practical and has good interlaboratory reproducibility.

Blood Chemical Analysis

Methodological issues in assessing reproducibility--a comparative study of various indices of reproducibility applied to repeat ELISA serologic tests for Lyme disease.

This study applies and evaluates a variety of different measures of reproducibility. As an example, repeat enzyme-linked immunosorbent assays (ELISA) for antibodies to Borrelia burgdorferi, the etiologic agent of Lyme disease, are used. These repeat tests were part of the 1991 Quality Control Assessment of provincial laboratories that was carried out by the Laboratory Centre for Disease Control (Ottawa). Twenty-seven sera from cases and non-cases were tested by three laboratories, and two laboratories repeated the tests once. Methodological issues discussed include: different methods of assessing reproducibility in the continuous scale; whether reproducibility should be assessed with data in continuous or categorical form; problems assessing the reproducibility of data that has been standardized using a calibration-regression line; and problems with external generalizability of reproducibility studies of rare diseases. The authors conclude that the statistical method used to assess the reproducibility of a test must be adjusted to suit individual study designs and data, and the usage of the test.

Antibodies, Bacterial

Measurement of ascorbic acid in human plasma and serum: stability, intralaboratory repeatability, and interlaboratory reproducibility.

We demonstrate that total ascorbic acid (TAA, the sum of ascorbic acid and dehydroascorbic acid) in properly prepared human plasma is stable at -70 degrees C for at least 6 years when preserved with dithiothreitol. TAA in human plasma or serum preserved with metaphosphoric acid degrades slowly, at the rate of no more than 1% per year. As assessed from our stability data and from data obtained from 23 laboratories over a period of > 2 years, the intralaboratory repeatability of TAA measurement is approximately 2 mumol/L, irrespective of TAA concentration. Nonchromatographic analytical methods involving dinitrophenylhydrazine and 0-phenylenediamine yield biased results relative to chromatographic methods. Within groups of laboratories that use roughly similar analytical methods, the interlaboratory measurement reproducibility CV for TAA is 15%.

Ascorbic Acid

Reframing Proteomics Measurement: Super Mass Spectrometry Framework and the Role of Delayed Electrospray Ionization Technique.

Dynamic range, repeatability, and reproducibility remain the central limitations of data-independent acquisition (DIA) proteomics. Current workflows emphasize protein group identification counts and throughput, but these metrics mask the fundamental measurement challenge: generating a repeatable, reproducible, high-fidelity, and relatively complete digital representation of complex proteomes. In particular, plasma proteomics spans more than 10 orders of magnitude in protein abundance, far exceeding the capacity and dynamic range of any single mass spectrometer. Incremental advances have not closed this gap. In this Perspectives article, I introduce the Super Mass Spectrometry framework and then highlight the Delayed Electrospray Ionization (Delayed-ESI) technique, as a practical approach to address these limitations. By producing compositionally identical but temporally staggered ion beams, the Delayed-ESI technique enables deterministic remeasurement of the same analyte profile, supporting various novel strategies to improve analytical figures of merit. While recent implementations of the Delayed-ESI technique have emphasized throughput, I argue that the broader value of the Delayed-ESI technique lies in extending dynamic range and improving repeatability and reproducibility&#x2500;objectives that should take precedence if proteomics is to evolve into a robust measurement science capable of supporting population-scale proteomics studies.

Proteomics

A statistical procedure for the estimation of accuracy parameters in interlaboratory studies.

Interlaboratory studies are conducted to estimate the accuracy of methods of laboratory measurements. The standard parameters used to describe this accuracy are the repeatability and the reproducibility. Usually variance components models are used to estimate these parameters. If the model assumptions are violated the resulting estimates for reproducibility and repeatability may, however, be biased. A new method of residual analysis in variance components models--developed by the author--may be used to detect violations of the model assumptions. If the residual analysis indicates that the model assumptions are violated, a simple robust method--which makes fewer assumptions--may be used for the estimation of accuracy parameters. The application of this residual analysis is demonstrated using data of an interlaboratory study. Graphical methods play an important role in the evaluation of the residuals. The analysis of the residuals uses methods similar to those used for the analysis of Studentized residuals in the linear model. The estimates obtained by the variance components model and the simple robust method are compared. The results of the residual analysis may be used to decide which of the two estimates can be considered more appropriate. The necessity of residual analysis in the analysis of interlaboratory studies by variance components models is pointed out. Potential hazards inherent to residual analysis in variance components models are discussed. Conclusions for the analysis of interlaboratory studies are drawn.

Bias

Rectal tone, distensibility, and perception: reproducibility and response to different distensions.

Increasing interest is focusing on the role of intestinal tone, distensibility, and mechanosensation in the genesis of abdominal symptoms. Experimental approaches usually feature balloon distension of the bowel with measurements of perception, tone, and compliance and/or elastance; however, the methodologies are standardized incompletely. We examined the reproducibility of repeated assessments of sensory perception, basal tone, and compliance and/or elastance of the rectum during distension. We also evaluated the response to inflations that varied in regard to control of pressure or volume, pattern of distension, and rate of inflation. Five healthy volunteers were studied under two separate protocols. The first featured a series of experiments on each of 5 days; the other consisted of 2 separate days of study. Repeated distensions evoked reproducible responses of sensation and compliance and/or elastance on a single day, providing a conditioning distension preceded them. Day-to-day variability was also sufficiently small to allow valid comparisons to be made on different days in healthy persons. The configuration of the distension profile (phasic, staircase, or ramp) and the rate of inflation (from 1 to 40 ml/s) had little effect on distensibility or perception. Perceptions were sometimes transient and sometimes constant, but no relationship was found between these temporal features and the magnitude of the stimulus. These observations help provide a basis as to how the responses to rectal distension can be best studied.

Adult