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PubMed1977

Evaluation of automated glucose oxidase methods for serum glucose: comparison to hexokinase of a colorimetric and an electrometric method.

Two automated glucose oxidase methods have been evaluated with respect to accuracy, precision, recovery, linearity and various potential interferences. Trinder's method on an AutoAnalyzer II had a between-day coefficient of variation (C.V.) of 2.6% (mean 228 mg/dl), was linear to 500 mg/dl, and produced a mean recovery of 99.7%. Comparison of Trinder's method with a manual, blanked hexokinase method yielded the regression equation: TR=--1.95 + HEX (1.04); Spearman's rho correlation coefficient was: 0.974. The Beckman System I glucose method had a between-day C.V. of 1.6% (mean 198 mg/dl), was linear to 500 mg/dl, and recovered an average of 98.0% of added glucose. Comparison with the same hexokinase method yielded: SYI = 1.27 + HEX (1.02: Spearman's rho = 0.991. None of the possible interfering compounds tested caused significant deviation of results by either method within the range of concentrations encountered physiologically. Trinder's method on the AutoAnalyzer II and the System I method are accurate, precise methods and are highly recommended for routine use in the clinical laboratory.

Autoanalysis↗

PubMed1993

Comparison of a cold enrichment method and the IDF method for isolation of Listeria monocytogenes from animal autopsy material.

The method most often used in Sweden for isolation of Listeria monocytogenes from animal autopsy material is a cold enrichment method. This method is very slow. The International Dairy Federation (IDF) has recently presented a method for detection of L. monocytogenes in milk and milk products that is complete in one week. During a two year period 69 specimens from dead animals with suspected listeriosis were examined for L. monocytogenes in parallel analyses with both the cold enrichment method and the IDF method. Samples derived from different autopsy material representing a variety of animals. L. monocytogenes was isolated in 27.5% of the samples with the IDF method but only in 4.3% with the cold enrichment method. It is concluded that the IDF method was more sensitive than the cold enrichment method.

Animals↗

PubMed2006

Comparison of an acetonitrile extraction/partitioning and "dispersive solid-phase extraction" method with classical multi-residue methods for the extraction of herbicide residues in barley samples.

An acetonitrile/partitioning extraction and "dispersive solid-phase extraction (SPE)" method that provides high quality results with a minimum number of steps and a low solvent and glassware consumption was published in 2003. This method, suitable for the analysis of multiple classes of pesticide residues in foods, has been given an acronymic name, QuEChERS, that reflects its major advantages (quick, easy, cheap, effective, rugged, safe). In this work, QuEChERS method, which was originally created for vegetable samples with a high amount of water, was modified to optimise the extraction of a wide range of herbicides in barley. Then, it was compared with known conventional multi-residue extraction procedures such as the Luke method, which was simplified and shortened by eliminating the Florisil clean-up (mini Luke) and the ethyl acetate extraction, which involves a subsequent clean-up by gel permeation chromatography (GPC) and which is the official extraction method used by some of European authorities. Finally, a simple acetone extraction was carried out to check the differences with the other three methods. Extracts were analysed by gas chromatography-time-of-flight mass spectrometry (GC-TOF/MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS). Mini Luke was significantly more effective for the extraction of non-polar and medium-polar compounds, but the best recoveries for polar compounds were achieved by QuEChERS and ethyl acetate methods. QuEChERS was the only method that provided an overall recovery value of 60-70% for non-, medium- and polar compounds, with some exceptions due to co-eluted matrix interferences. Clean-up by dispersive SPE was effective and did not differ so much with ethyl acetate extracts considering that QuEChERS clean-up step is much easier and less time-consuming. As a conclusion, it resulted to be the most universal extraction method by providing a well-defined phase separation without dilution and achieving acceptable recoveries in average including the extraction of the always difficult acidic herbicides. However, recoveries were not as good as required for validation purposes suggesting that residues are prone to strong matrix interactions in dry samples as barley and further method adaptation incrementing solvent strength, extraction time or more acidic or basic conditions is needed in order to achieve a complete extraction.

Acetonitriles↗

PubMed2005

A comparison of the rhizosphere-based method with DTPA, EDTA, CaCl2, and NaNO3 extraction methods for prediction of bioavailability of metals in soil to barley.

A rhizosphere-based method was compared with DTPA, EDTA, CaCl2, and NaNO3 extraction methods for the evaluation of bioavailability of heavy metals in soil to barley. The extractable amounts of Cr, Cu, Zn and Cd analyzed by the rhizosphere-based method correlated significantly with the metal content of barley roots. The extractable metals identified by DTPA, EDTA, CaCl2 and NaNO3 methods exhibited relatively poor or no correlation with the metal content of barley roots. The stepwise multiple regression equation of the rhizosphere-based method was the simplest one, as no soil properties needed to be entered, whereas the equations for the DTPA, EDTA, CaCl2 and NaNO3 extraction methods always require those variables. The most distinct feature of the rhizosphere-based method was that the proposed method was suitable for acidic, neutral and near alkaline soils. In contrast, the other extraction methods were restricted to soil types. In summary, the rhizosphere-based method is the most robust approach for evaluation of bioavailability of metals in soil to barley.

Biological Availability↗

PubMed1981

Comparison of an agarose electrophoresis method and four ion-exchange methods for the measurement of the creatine kinase MB fraction in human serum.

Five CK-MB methods were compared; Corning agarose gel electrophoresis, Worthington CPK Isoenzyme columns, E-C columns, Lancer MB/CPK Isoenzyme, and our own DEAE-cellulose column method. The best precision was obtained with agarose electrophoresis at elevated CK-MB levels. However the agarose electrophoresis method was less sensitive than any of the ion-exchange methods. The detection limit for CK-MB by agarose electrophoresis was variable in the range of 67-17 U/L. A significant bias was shown to exist between the ion-exchange methods and the agarose electrophoresis method. Results by the latter were on the average 4.3, 3.6, 2.7 and 1.4 times greater than CK-MB measurements obtained by the Lancer, E-C, Worthington, and DEAE-cellulose methods, respectively. The bias observed between methods was largely due to the low recoveries of CK-MB activity with the ion-exchange methods; recoveries of 106-122%, 68-78%, 42-44%, 16-19%, and 13-14% were obtained at levels of 18-162 U/L, with the agarose electrophoresis, DEAE-cellulose, Worthington E-C, and Lancer methods, respectively.

Chromatography, Ion Exchange↗

PubMed1997

Novel method to investigate kinetics of rat skin cells by means of an occlusive dressing method using bromodeoxyuridine.

We developed a novel technique to detect S-phase skin cells by applying bromodeoxyuridine (BrdU) epicutaneously using an occlusive dressing (OD) method. BrdU was scarcely absorbed from the skin with a simple epicutaneous application, whereas the incorporation of BrdU was very well promoted with the use of our OD method. We applied BrdU on the backs of rats using this method and investigated the conditions required for an optimal response, with a special focus on the period of application, the concentration of BrdU used and vehicles suitable for the immunocytochemical staining of this agent. From these experiments, we were able to determine that an application time of at least 60 min was necessary to liable S-phase cells, a 2% concentration of BrdU was needed to obtain consistent labeling and aqueous vehicles are satisfactory solvents for BrdU preparations. Epidermal keratinocytes and S-phase cells in the upper portion of dermis were clearly labeled after either intraperitoneal injection of BrdU or after administration by means of the OD method. To ascertain whether this latter method could provide an effective alternative to intraperitoneal injection, we compared the labeling patterns of both methods with respect to the speed of migration of BrdU-labeled basal cells from the basal layer to the horny layer of epidermis. Using either of these two methods, basal keratinocytes were labeled immediately after administration. Three days after the first administration, BrdU-labeled cells were detected in the middle layer of the epidermis, but after 8 days, they were no longer evident in epidermal tissue. As another means of comparing both methods, we used antibody to proliferating cell nuclear antigen (PCNA) and compared the ratio of PCNA-positive basal cells to BrdU-labeled basal cells. The number of PCNA-positive cells was about 4.6 times greater than the number of BrdU-labeled basal cells by both methods. We concluded that the OD method could be used as a substitute for intraperitoneal injection in order to observe cell kinetics using bromodeoxyuridine.

Administration, Topical↗

PubMed1989

Prediction of imipramine serum levels in enuretic children by a Bayesian method: comparison with two other conventional dosing methods.

The aim of the present study was to characterize the kinetic behavior of imipramine (IMI) and desipramine in enuretic children and to evaluate the performance of different methods for dosage prediction based on individual and/or population data. The study was carried out in 135 enuretic children (93 boys) ranging in age between 5 and 13 years undergoing treatment with IMI in variable single doses (25-75 mg/day) administered at night. Sampling time was one-half the dosage interval at steady state. The number of data available for each patient varied (1-4) and was essentially limited by clinical criteria. Pharmacokinetic calculations were performed using a simple proportional relationship (method 1) and a multiple nonlinear regression program (MULTI 2 BAYES) with two different options: using the ordinary least-squares method (method 2) and the least-squares method based on the Bayesian algorithm (method 3). The results obtained point to a coefficient of variation for the level/dose ratio of the drug (58%) that is significantly lower than that of the metabolite (101.4%). The forecasting capacity of method 1 is deficient both regarding accuracy [mean prediction error (MPE) = -5.48 +/- 69.15] and precision (root mean squared error = 46.42 +/- 51.39). The standard deviation of the MPE (69) makes the method unacceptable from the clinical point of view. The more information that is available concerning the serum levels, the greater are the accuracy and precision of methods (2 and 3). With the Bayesian method, less information on drug serum levels is needed to achieve clinically acceptable predictions.

Adolescent↗

PubMed1990

Evaluation of test sensitivity and test specificity of three field methods for quantification of chloroquine in urine using liquid chromatography as reference method.

We have evaluated three quantitative colorimetric methods [bromthymol blue (BTB), Haskins, and Saker-Salomons (S-S)] for measurement of concentrations of chloroquine (CQ) in urine. A graphical evaluation of test sensitivity and test specificity at different concentrations of CQ in authentic urine samples was used. Liquid chromatography (LC) was used as a reference method. The Haskins method showed the highest test specificity (89%) and test sensitivity (97%), at a discrimination value of 2 mumols/L. At the same sensitivity (97%) as the Haskins method, S-S and BTB methods had test specificities of 49 and 13%, respectively. The correlation coefficients between the LC method and the colorimetric methods were all higher than 0.94 at a urinary concentration range of 2-100 mumols/L. The intra- and interassay variations for the colorimetric methods were all uniformly less than 15% at 2 mumols/L, but the Haskins method showed less variation than the other two methods.

Child↗

PubMed2005

The cuff method: a pilot study of a new method of monitoring neuromuscular function.

BACKGROUND: A new method of monitoring neuromuscular blockade based on a modified blood pressure cuff that incorporates stimulating electrodes was compared with mechanomyography (MMG) ('gold standard'). METHODS: Forty adults and 20 children (ASA I-II) underwent neuromuscular blockade monitoring on the contralateral arms and on the same arm using the new cuff method and MMG. Only train-of-four (TOF) ratios > 0.1 and T(1) heights > 0 were studied. Supramaximal stimulation was also assessed. A device based on a PC with an analogue-to-digital conversion card was used to control and synchronize MMG and the cuff method. The agreement between both methods was assessed using the statistical method of Bland and Altman. RESULTS: When TOF ratios were >0.7, the bias between the two methods was -0.04 with the limits of agreement ranging from -0.21 to -0.12 (95% CI -0.06 to -0.02). The T(1) > 0 heights bias was -0.01 with the limits of agreement ranging from -0.26 to 0.24 (95% CI -0.02 to -0.003). The sensitivity of the cuff method was 88%, with a specificity of 85% and an accuracy of 86%. CONCLUSION: This pilot study indicates that the cuff method could be useful to monitor neuromuscular blockade according to the bias and limits of agreement compared with MMG, particularly when the degree of blockade was evaluated by TOF ratios > 0.7. The new cuff method is easy and simple to use. However, further studies in a larger number of patients are necessary to confirm these favourable preliminary results.

Adolescent↗

PubMed2004

A comparison of the traditional method of counting viable cells and a quick microplate method for monitoring the growth characteristics of Listeria monocytogenes.

AIMS: To determine: (i) the growth parameters (specific growth rate, lag time, asymptotic amount of growth, generation time and time for maximum growth rate) of Listeria monocytogenes in different broths by standard cultivation methods and (ii) whether a microplate method in conjunction with a standard nondedicated plate reader could be adapted to routine assay. METHODS AND RESULTS: Growth curves were determined from cell numbers in a standard tube method at 2 h intervals by serial dilution and plating, and in a microplate method by absorbance measurements. Growth curves were fitted with a modified Gompertz function. CONCLUSIONS: The microplate method was similar to the standard cultivation methods in accuracy, required less chemical reagents, and considerably reduced the time required for analyses. This work also illustrates that growth characteristics of bacteria are not necessarily constant, and depend on the methodology used. SIGNIFICANCE AND IMPACT OF THE STUDY: It is not the intended purpose of this paper to present all the data for the media tested but instead to illustrate the success of the microplate method for studying growth kinetics compared to a standard cultivation method and system precision. The method will be of considerable benefit to laboratories unable to afford dedicated workstations.

Bacteriological Techniques↗

PubMed1996

Comparison of a spectrophotometric microdilution method with RPMI-2% glucose with the National Committee for Clinical Laboratory Standards reference macrodilution method M27-P for in vitro susceptibility testing of amphotericin B, flucytosine, and fluconazole against Candida albicans.

The National Committee for Clinical Laboratory Standards has proposed a reference broth macrodilution method for in vitro antifungal susceptibility testing of yeasts (the M27-P method). This method is cumbersome and time-consuming and includes MIC endpoint determination by visual and subjective inspection of growth inhibition after 48 h of incubation. An alternative microdilution procedure was compared with the M27-P method for determination of the amphotericin B, flucytosine, and fluconazole susceptibilities of 8 American Type Culture Collection strains (6 of them were quality control or reference strains) and 50 clinical isolates of candida albicans. This microdilution method uses as culture medium RPMI 1640 supplemented with 18 g of glucose per liter (RPMI-2% glucose). Preparation of drugs, basal medium, and inocula was done by following the recommendations of the National Committee for Clinical Laboratory Standards. The MIC endpoint was calculated objectively from the turbidimetric data read at 24 h. Increased growth of C. albicans in RPMI-2% glucose and its spectrophotometric reading allowed for the rapid (24 h) and objective calculation of MIC endpoints compared with previous microdilution methods with standard RPMI 1640. Nevertheless, good agreement was shown between the M27-P method and this microdilution test. The MICs obtained for the quality control or reference strains by the microdilution method were in the ranges published for those strains. For clinical isolates, the percentages of agreement were 100% for amphotericin B and fluconazole and 98.1% for flucytosine. These data suggest that this microdilution method may serve as a less subjective and more rapid alternative to the M27-P method for antifungal susceptibility testing of yeasts.

Amphotericin B↗

PubMed1990

Blinded comparison of a direct immunofluorescent monoclonal antibody staining method and a Giemsa staining method for identification of Pneumocystis carinii in induced sputum and bronchoalveolar lavage specimens of patients infected with human immunodeficiency virus.

A new direct immunofluorescence monoclonal antibody (DFA) method (Genetic Systems, Inc., Seattle, Wash.) for identification of Pneumocystis carinii in induced sputum and bronchoalveolar lavage specimens was compared in a blinded study with an established Giemsa stain method. We evaluated 148 consecutive clinical specimens from 104 patients with the following results. For the 67 patients (64%) infected with the human immunodeficiency virus (HIV), 49 were initially negative by both the DFA and the Giemsa methods, none were negative by DFA and positive by Giemsa, 6 were positive by DFA and negative by Giemsa, and 12 were positive by both methods, for a sensitivity and a negative predictive value of greater than 99%. For the six patients positive by DFA and negative by Giemsa, all were positive by both methods on evaluation of subsequently obtained clinical specimens, suggesting a specificity of greater than 99% and a false-positive rate of less than 1%. For 37 patients whose HIV status was negative or unknown, 35 were negative by both methods and 2 were positive by DFA and negative by Giemsa. The DFA method was simple to perform and required less time for scoring of stained slides than the Giemsa method, but care had to be taken to avoid false-positive readings due to extraneous fluorescence. This study indicates that the DFA method represents an advance as a sensitive, simple, and rapid way to identify P. carinii in induced sputum and bronchoalveolar lavage specimens from HIV-infected patients and suggests greater sensitivity of the DFA than the Giemsa method in this patient population.

Antibodies, Monoclonal↗

PubMed1999

Comparison of two new methods for the measurement of lung volumes with two standard methods.

BACKGROUND: The two most commonly used methods for the measurement of lung volumes are helium dilution and body plethysmography. Two methods have been developed which are both easier and less time consuming to perform. Mathematical modelling uses complex calculations from the flow-volume loop to derive total lung capacity (TLC), and the nitrogen balance technique uses nitrogen from the atmosphere to calculate lung volume in a similar way to helium dilution. This study was designed to compare the two new methods with the two standard methods. METHODS: Sixty one subjects were studied, 23 with normal lung function, 17 with restrictive airway disease, and 21 with obstructive ventilatory defects. Each subject underwent repeated measurements of TLC by each of the four methods in random order. Reproducible values were obtained for each method according to BTS/ARTP guidelines. Bland-Altman plots were constructed for comparisons between the methods and paired t tests were used to assess differences in means. RESULTS: Bland-Altman plots showed that the differences between body plethysmography and helium dilution fell into clinically acceptable ranges (agreement limits +/-0.9 l). The agreement between mathematical modelling or the nitrogen balance technique and helium dilution or body plethysmography was poor (+/-1.8-3.4 l), especially for subjects with airflow obstruction. CONCLUSIONS: Neither of the new methods agrees sufficiently with standard methods to be useful in a clinical setting.

Adult↗

PubMed1999

Administration of the BCG vaccination using the multipuncture method in schoolchildren: a comparison with the intradermal method.

BACKGROUND: BCG vaccination using the multipuncture device (the Heaf gun) is recommended in the UK for infants and very small children only. The aim of this study was to investigate the rate of conversion of the tuberculin test, the safety and acceptability of BCG vaccination using the multipuncture device and to compare it with the conventional intradermal method in schoolchildren. METHODS: Schoolchildren attending schools in Tower Hamlets who were eligible for BCG vaccination were tuberculin tested using the Heaf gun. Those with grade 0-1 reaction were randomised to receive BCG vaccination using either the multipuncture or the intradermal method. The site of BCG vaccination was inspected after eight weeks for inflammatory changes and scarring. A questionnaire about pain and inflammation at the site of vaccination was completed. The Heaf test was repeated at eight weeks and its results were assessed by an examiner unaware of the results of the previous Heaf test and the method of BCG administration. The Heaf test conversion was deemed to have occurred if there was a change of at least one grade in the response. RESULTS: One hundred and sixty nine children (83 girls) of mean age 11.8 years completed the study, of which 81 received BCG by the multipuncture method. The Heaf test did not convert in 22 of 81 (27. 2%) receiving BCG by the multipuncture device compared with six of 88 (6.8%) who received the vaccine by the intradermal method (odds ratio 0.2, 95% confidence interval 0.07 to 0.55). The BCG scar was visible in all children who had intradermal BCG compared with 67 of 81 (81.8%) of the multipuncture group. The multipuncture method was less painful and caused fewer inflammatory changes than the intradermal method. CONCLUSIONS: In schoolchildren the multi-puncture device for administering BCG caused a lower rate of tuberculin conversion as measured by the Heaf test and less of an inflammatory response than the intradermal method. The method needs to be modified before it is applied on a wider scale to schoolchildren.

Adolescent↗

PubMed1997

An improved method for cultivation of Mycobacterium paratuberculosis from bovine fecal samples and comparison to three other methods.

A new method (NADC) for isolation of Mycobacterium paratuberculosis from fecal samples is described and evaluated using fecal samples from a known paratuberculosis-infected herd of cattle. The NADC method includes centrifugation of the total fecal sample supernatant and use of a 2-step decontamination protocol. The growth rate of M. paratuberculosis and contamination rate of cultures when using this method are compared to 3 other published methods: sedimentation, centrifugation, and Cornell. Sensitivity was lowest for the Cornell method precluding detection of some low shedders; however, contamination was not observed for this method. Contamination was the most severe in samples processed by the centrifugation method but was also high for the sedimentation method, resulting in unreadable culture tubes for some fecal samples. The NADC method was 10-fold more sensitive for detection of M. paratuberculosis colonies and contamination was significantly reduced compared to other 3 methods.

Animals↗

PubMed2006

A method for calculating the exact angle required during pedicle subtraction osteotomy for fixed sagittal deformity: comparison with the trigonometric method.

OBJECTIVE: Pedicle subtraction osteotomy (PSO) has emerged as a powerful procedure for correcting fixed sagittal deformity. There has only been one attempt to quantify the magnitude of correction needed to restore sagittal balance; the trigonometric method for calculating the desired PSO angle is an approximation. We propose a method for calculating the exact angle required for PSO and explore how this angle differs from that obtained via the trigonometric method in illustrative cases. METHODS: We conducted a mathematical analysis of the spine with application in illustrative cases. The trigonometric method calculates the necessary angular correction at the axial level of the PSO, but along the sacral vertical line. However, the angular measurement should take place at the true axis of rotation, the apex of the PSO. Measurements were taken from full-length standing x-rays, and both methods were explored. RESULTS: The trigonometric method for calculating PSO angle is an exact measurement only if the apex of the PSO site lies on the sacral vertical line. As the apex of the PSO site moves anterior to that line, the trigonometric approximation underestimates the actual angle. As the apex of the PSO moves posterior to that line, the trigonometric approximation overestimates the actual angle. CONCLUSION: The trigonometric method for calculating the PSO angle required for surgical deformity correction is an approximation, but its validity in clinical practice was confirmed by this study. The exact angle is obtained by a method centered on the apex of the PSO site. Although the difference between these angles is small, it is an important conceptual point for spine surgeons. Measurement of the exact angle is easily performed and should replace the trigonometric method for calculating the required PSO angle when standard digital measurement tools are available.

Aged↗

PubMed1994

Comparison of four methods for calculating diffusing capacity by the single breath method.

In 283 patients referred for testing in an outpatient pulmonary function laboratory, we studied the single-breath diffusing capacity of the lungs for carbon monoxide (Dco) using the Ogilvie (Og), Jones-Meade (JM), Epidemiological Standardization Project (ESP), and three-phase iterative methods (3PIT, similar to the three equation method). The Dco maneuvers were performed using automated equipment and American Thoracic Society (ATS) recommended procedures. There were small but significant differences in mean Dco, the ESP method yielding the largest, followed in order by JM, 3PIT, and Og methods. The 3PIT and JM Dcos were in close agreement in all degrees and patterns of pulmonary function abnormality. The Og Dco method was 6 percent less than JM in patients with normal pulmonary function, although the difference was less in patients with expiratory flow limitation, restriction, or reduced Dco. There were no differences in the reproducibility of Dco measurements among the methods. Based on these results and a review of the literature, we conclude the following: (1) when measuring single-breath Dco using automated equipment that follows ATS recommended procedures for collecting a single expired gas sample of 500-ml volume, calculated Dco is largest using ESP method, following by JM, 3PIT, and Og methods; (2) in patients with reduced Dco associated with obstructive or restrictive abnormalities, the Og, 3PIT, and JM timing methods are nearly equivalent; and (3) reproducibility of Dco is the same by all methods.

Airway Obstruction↗

PubMed2005

Method performance study of the determination of total nitrogen in soy sauce by the Kjeldahl method.

The objective of this collaborative study was to evaluate the proposed method for determining the total nitrogen in soy sauce by the Kjeldahl method submitted to the Codex Alimentarius Commission for endorsement in accordance with the protocol for the design, conduct, and interpretation of method-performance studies. The digestive conditions of the proposed method are the addition of 10 mL of H2SO4, 10 g (8 g by using a block digester) of K2SO4, and 1 mL of 20% CuSO4 x 5H2O and 80 min boiling period after the liquid is cleared by a heating device. Seventeen laboratories participated, analyzing five soy sauce samples as blind duplicates. Since the volume sampling method used in the JAS (Japanese Agricultural Standard) method showed lower accuracy of data because of the density of soy sauce, the method of sampling by weight was adopted as the proposed method. The total amount of outlier data was within acceptable limits for method-performance studies (< or = 22.2%). Lysine and ammonium sulfate recoveries for all laboratories were > or = 98% and > or = 99% respectively. The RSDr (repeatability relative standard deviation) values ranged from 0.4 to 1.3%, and the RSDR (reproducibility relative standard deviation) values were from 0.8 to 1.9%. HORRAT (RSDR/predicted RSDR) for the reproducibility showed 0.2 to 0.4, indicating acceptable precision of the method and excellent analytical performance.

Food Analysis↗