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Rapid automated antimicrobial susceptibility testing of Streptococcus pneumoniae by use of the bioMerieux VITEK 2.

The VITEK 2 is a new automated instrument for rapid organism identification and susceptibility testing. It has the capability of performing rapid susceptibility testing of Streptococcus pneumoniae with specially configured cards that contain enriched growth medium and antimicrobial agents relevant for this organism. The present study compared the results of testing of a group of 53 challenge strains of pneumococci with known resistance properties and a collection of clinical isolates examined in two study phases with a total of 402 and 416 isolates, respectively, with a prototype of the VITEK 2. Testing was conducted in three geographically separate laboratories; the challenge collection was tested by all three laboratories, and the unique clinical isolates were tested separately by the individual laboratories. The VITEK 2 results of tests with 10 antimicrobial agents were compared to the results generated by the National Committee for Clinical Laboratory Standards reference broth microdilution MIC test method. Excellent interlaboratory agreement was observed with the challenge strains. The overall agreement within a single twofold dilution of MICs defined by the VITEK 2 and reference method with the clinical isolates was 96.3%, although there were a number of off-scale MICs that could not be compared. The best agreement with the clinical isolates was achieved with ofloxacin and chloramphenicol (100%), and the lowest level of agreement among those drugs with sufficient on-scale MICs occurred with trimethoprim-sulfamethoxazole (89.7%). Overall there were 1.3% very major, 6.6% minor, and no major interpretive category errors encountered with the clinical isolates, although >80% of the minor interpretive errors involved only a single log(2) dilution difference. The mean time for generation of susceptibility results with the clinical isolates was 8.1 h. The VITEK 2 provided rapid, reliable susceptibility category determinations with both the challenge and clinical isolates examined in this study.

Anti-Bacterial Agents↗

Susceptibility of filamentous fungi to voriconazole tested by two microdilution methods.

The growing number of fungal infections, coupled with emerging resistance to classical antifungal agents, has led to the development of new agents, among them voriconazole. Susceptibility to voriconazole was tested by two microdilution techniques: the National Committee for Clinical Laboratory Standards reference method M38-A and a colorimetric method, Sensititre YeastOne. The study tested a total of 244 isolates: 223 Aspergillus (136 Aspergillus fumigatus, 37 A. niger, 26 A. terreus, 16 A. flavus, 7 A. nidulans, and 1 A. ustus), 14 Fusarium (8 Fusarium moniliformis, and 6 F. oxysporum), 6 Scedosporium apiospermum, and 1 Rhizomucor pusillus strain and four control strains (Candida parapsilosis ATCC 22019, C. krusei ATCC 6258, A. fumigatus ATCC 204305, and A. flavus ATCC 204304). For all tested species except one F. moniliforme strain and R. pusillus, the MIC, the MIC at which 50% of the isolates are inhibited (MIC(50)), and MIC(90) ranges of </=1 microg/ml were obtained for voriconazole, indicating excellent activity against these species. The high rate of agreement between the two methods used (97 to 99%) suggests that the Sensititre YeastOne colorimetric method may be a valuable tool for determining the susceptibility of filamentous fungi to voriconazole.

Antifungal Agents↗

Bone invasion in patients with oral cavity cancer: comparison of conventional CT with PET/CT and SPECT/CT.

PURPOSE: To prospectively compare the accuracy of helical contrast material-enhanced computed tomography (CT) with that of CT and positron emission tomography (PET) combined and CT and single photon emission CT (SPECT) combined in the detection of bone invasion in patients scheduled to undergo surgery for clinically suspected oral cavity carcinoma with possible bone invasion, with surgical results as the reference standard. MATERIALS AND METHODS: This study had local ethical committee approval, and all patients gave written informed consent. Thirty-four consecutive patients (17 men, 17 women; mean age, 64.2 years; age range, 46.0-84.6 years) who were clinically suspected of having bone invasion from oral cavity carcinoma prospectively underwent helical contrast-enhanced CT, coregistered PET/CT, and coregistered SPECT/CT. Two radiologists assessed the contrast-enhanced CT images and two nuclear medicine physicians separately assessed the PET/CT and SPECT/CT images in consensus and without knowledge of the results of other imaging tests. The presence of bone involvement as suggested with an imaging modality was compared with histologic findings in the surgical specimen. RESULTS: With histologic findings as the standard of reference, the accuracy of SPECT/CT (88% [30 of 34 patients]) was lower than that of PET/CT and contrast-enhanced CT (94% [32 of 34 patients] and 97% [33 of 34 patients], respectively). Sensitivity was highest with PET/CT (100% [12 of 12 patients]), and specificity was highest with contrast-enhanced CT (100% [22 of 22 patients]). Fluorine 18 fluorodeoxyglucose (FDG) uptake seen on two sides of the same cortical bone was not a helpful imaging pattern for better identifying bone invasion in patients without evident cortical erosion on CT scans. CONCLUSION: The assessment of cortical erosion with contrast-enhanced CT and the CT information from PET/CT are the most reliable methods for detecting bone invasion in patients with oral cavity carcinoma. FDG uptake seen on PET/CT images does not improve identification of bone infiltration.

Aged↗

Cartilaginous defects of the femorotibial joint: accuracy of coronal short inversion time inversion-recovery MR sequence.

PURPOSE: To retrospectively assess the diagnostic performance of the short inversion time inversion-recovery (STIR) magnetic resonance (MR) sequence for depiction and classification of articular cartilaginous lesions in femorotibial joint, with arthroscopy as reference standard. MATERIALS AND METHODS: Institutional review board did not require approval and informed consent for review of patients' records or images. All patients (and parents of underage patients) agreed to use of their data. Two musculoskeletal radiologists independently analyzed femorotibial cartilage on coronal STIR images from 84 knee MR examinations in 83 patients (48 male patients [49 knees], 35 female patients; mean age, 39.5 years). Slightly modified Outerbridge classification was used: grade 0, normal cartilage; grade 1, softening or swelling; grade 2, partial-thickness defect; grade 3, fissuring to the level of the subchondral bone; and grade 4, exposed subchondral bone. Arthroscopy performed within 15 weeks was the standard of reference. Classification for arthroscopy differed only in definition of grade 1 (softening or swelling of cartilage). Sensitivity, specificity, accuracy, positive and negative predictive values, and weighted kappa values were calculated to assess interobserver reliability. RESULTS: At arthroscopy, 212 (63%) of 336 surfaces were classified as grade 0 (normal); 37 (11%), as grade 1 abnormalities; 30 (9%), as grade 2 lesions; 25 (7%), as grade 3 lesions; and 32 (10%), as grade 4 lesions. Grades 0 and 1 were considered normal; grades 2-4, as abnormal. For detection of contour defects of the cartilaginous surface, coronal STIR MR imaging had sensitivity values of 77% and 76%, specificity values of 96% and 89%, accuracy values of 91% and 85%, positive predictive values of 86% and 70%, and negative predictive values of 92% and 91% for readers 1 and 2, respectively. Weighted kappa value was 0.63. CONCLUSION: Contour defects of femorotibial cartilage can be detected with reasonable accuracy with routine STIR sequence.

Adolescent↗

Uterine fibroids: contrast-enhanced MR angiography to predict ovarian artery supply--initial experience.

PURPOSE: To prospectively evaluate the diagnostic performance of contrast material-enhanced magnetic resonance (MR) angiography in helping predict ovarian artery supply of uterine fibroids by using postembolization conventional aortography as the reference standard. MATERIALS AND METHODS: The protocol for the study was approved by the institutional review board, and each patient gave informed consent. Ninety consecutive women (mean age, 42.5 years; range, 33-63 years) underwent MR angiography before uterine artery embolization (UAE). The number and origin of the ovarian arteries were determined. Ovarian artery supply of fibroids was graded as very unlikely, possible, or very likely by using a scoring system based on a combination of MR angiographic findings. MR angiographic results were compared with those of conventional aortography performed after UAE in all patients and followed by selective angiography in case of a suspected ovarian artery supply of fibroids. Analysis of the association between MR angiographic grading and conventional angiography as the standard of reference was performed with a chi(2) trend test. Sensitivity and specificity, including exact 95% confidence intervals (CIs), of MR angiography were determined. RESULTS: MR angiography depicted 18 ovarian arteries (four bilateral, 10 unilateral), one with an atypical origin. Five ovarian arteries were classified as very likely; three, as possible; and 10, as very unlikely sources of arterial fibroid supply. Seven (39%) of 18 ovarian arteries detected at MR angiography were visible at conventional aortography. Fibroid supply was verified at selective angiography in five ovarian arteries in five (6%) of 90 patients. There was a strong association between MR angiographic grading and the results of conventional angiography (P = .002). Sensitivity of MR angiography in depicting ovarian artery supply (grade, possible or very likely) was 100% (five of five, 95% CI: 48%; 100%) and specificity was 77% (10 of 13, 95% CI: 46%; 95%). CONCLUSION: Contrast-enhanced MR angiography can help predict ovarian artery supply of uterine fibroids.

Adult↗

Preoperative diagnosis of carotid artery stenosis: accuracy of noninvasive testing.

BACKGROUND AND PURPOSE: Carotid endarterectomy has been shown to be beneficial in symptomatic patients with a severe stenosis (70% to 99%) of the internal carotid artery (ICA). Digital subtraction angiography (DSA) is the standard of reference in the diagnosis of carotid artery stenosis but has a relatively high complication rate. In a diagnostic study we investigated the accuracy of noninvasive testing compared with DSA. METHODS: In a prospective diagnostic study we performed duplex ultrasound (DUS), magnetic resonance angiography (MRA), and DSA on 350 consecutive symptomatic patients. Stenoses were measured with the observers blinded for clinical information and other test results. Separate and combined test results of DUS and MRA were compared with the reference standard DSA. Only the stenosis measurements of the arteries on the symptomatic side were included in the analyses. RESULTS: DUS analyzed with previously defined criteria resulted in a sensitivity of 87.5% (95% CI, 82.1% to 92.9%) and a specificity of 75.7% (95% CI, 69.3% to 82.2%) in identifying severe ICA stenosis (70% to 99%). Stenosis measurements on MRA yielded a sensitivity of 92.2% (95% CI, 86.2% to 96.2%) and a specificity of 75.7% (95% CI, 68.6% to 82.5%). When we combined MRA and DUS results, agreement between these 2 modalities (84% of patients) gave a sensitivity of 96.3% (95% CI, 90.8% to 99.0%) and a specificity of 80.2% (95% CI, 73.1% to 87.3%) for identifying severe stenosis. CONCLUSIONS: MRA showed a slightly better accuracy than DUS in the diagnosis of carotid artery stenosis. To achieve the best accuracy, however, both tests should be performed subsequently.

Adult↗

Accuracy of whole-body dual-modality fluorine-18-2-fluoro-2-deoxy-D-glucose positron emission tomography and computed tomography (FDG-PET/CT) for tumor staging in solid tumors: comparison with CT and PET.

PURPOSE: To assess the accuracy of positron emission tomography/computed tomography (PET/CT) when staging different malignant diseases. PATIENTS AND METHODS: This was a retrospective, blinded, investigator-initiated study of 260 patients with various oncological diseases who underwent fluorine-18-2-fluoro-2-deoxy-d-glucose PET/CT for tumor staging. CT images alone, PET images alone, PET + CT data viewed side by side, and fused PET/CT images were evaluated separately according to the tumor-node-metastasis system. One hundred forty patients with tumors not staged according to the tumor-node-metastasis system or a lack of reference standard were excluded from data analysis; 260 patients were included. Diagnostic accuracies were determined for each of the four image sets. Histopathology and a clinical follow-up of 311 (+/- 125) days served as standards of reference. RESULTS: PET/CT proved significantly more accurate in assessing tumor-node-metastasis system stage compared with CT alone, PET alone, and side-by-side PET + CT (P < .0001). Of 260 patients, 218 (84%; 95% CI, 79% to 88%) were correctly staged with PET/CT, 197 (76%; 95% CI, 70% to 81%) with side-by-side PET + CT, 163 (63%; 95% CI, 57% to 69%) with CT alone, and 166 (64%; 95% CI, 58% to 70%) with PET alone. Combined PET/CT had an impact on the treatment plan in 16, 39, and 43 patients when compared with PET + CT, CT alone, and PET alone, respectively. CONCLUSION: Tumor staging with PET/CT is significantly more accurate than CT alone, PET alone, and side-by-side PET + CT. This diagnostic advantage translates into treatment plan changes in a substantial number of patients.

Adolescent↗

Reference materials and reference methods in clinical chemistry.

In clinical chemistry today there are often several different methods and instruments in use for the analysis of a particular constituent. When these different methods and instruments are used for careful analysis of the same control specimen, as in assigned value determinations, the assigned values obtained may be significantly different, and these differences may be of clinical importance. In order to determine the reasons for such differences, reference systems or reference points are needed. The reference systems for calibration (calibration materials, standard reference materials) and control (control materials), where the matrices of these different materials must have quite different characteristics, are referred to jointly as reference materials. In order to compare methods, reference methods are needed with known, high reliability. Because of the amount of specimen material needed, the time needed for analysis and the facilities required, reference methods are not suitable for routine analysis. IDeally, standard reference materials are developed first, followed by a definitive method, which is then used to evaluate a candidate reference method. This is currently the case for only a small number of the constituents analyzed in the clinical chemistry laboratory. In this paper the characteristics of the different kinds of reference materials and of reference methods are described. The differences in terminological usage found in the literature are discussed. Limitations of knowledge and technique may necessitate certain compromises, with respect to the ideal characteristics of reference materials and reference methods. Possible compromises are discussed, and the various sources of error associated with them are pointed out. However, these compromises can also lead to improvement in the reliability and comparability of analytical results from different laboratories. As an example of such improvement, the results are presented for assigned value determinations by highly qualified reference laboratories on control specimens for interlaboratory surveys; the decision limits are included and the results are compared with those of the survey participants. Thus various ways are indicated for the use of reference materials and reference methods to improve the reliability and comparability of analytical results, suited to the current state of the art.

Animals↗

A robust liquid chromatography tandem mass spectrometry method for total plasma homocysteine determination in clinical practice.

BACKGROUND: Total plasma homocysteine has emerged as an independent risk factor for vascular disease. To meet increasing requests by clinicians for this homocysteine determination, a rapid assay for a routine use has been developed. METHODS: A robust, stable, isotope-dilution liquid chromatography tandem mass spectrometry (LC/MS/MS) method is described, including all the practical details and analytical performance results. RESULTS: The method allows homocysteine quantitation over a linear working range up to 100 micromol/L, with the limit of quantification estimated at a low value of 0.09 micromol/L. Total analytical imprecision is less than 4%. Accuracy was assessed by measuring the homocysteine concentration in a serum Standard Reference Material. CONCLUSIONS: The method was demonstrated to be quick, reliable and cheap after 1 year of use on a time-shared instrument in our hospital unit.

Chromatography, High Pressure Liquid↗

Isotope ratio analysis of lead in biological materials by inductively coupled plasma mass spectrometry.

Inductively coupled plasma mass spectrometry (ICP-MS) allowed 0.2-0.3% imprecision (1 sigma) in 204Pb/206Pb, 207Pb/206Pb, and 208Pb/206Pb measurements at the 20-100 ppb level, which was precise enough to detect some of the isotopic variations observed in nature. Mass discrimination could be corrected within +/- 0.5% of the true value by periodical analysis of standard reference material of known lead isotopic composition. As a separation method for lead in human bone, which contains enormous amounts of calcium and phosphorus, anion exchange of the Pb-Br complex was found to be effective. Lead isotope ratios in bone, measured by ICP-MS after separation, were consistent with those measured by thermal ionization mass spectrometry. Hair matrix did not have any influence on the accuracy and precision of the analysis; a digested sample could be directly analyzed and this offered rapid sample throughput. Preliminary data on lead isotope ratios in bone and hair from prehistoric and contemporary Japanese are presented.

Archaeology↗

A simple and rapid spectrophotometric determination of thallium(III) with trifluoperazine hydrochloride.

A simple, sensitive and rapid spectrophotometric method was developed for the determination of thallium(III) using trifluoperazine hydrochloride (TFPH). The method is based on the oxidation of TFPH by thallium(III) in a phosphoric acid medium to form a red-colored radical cation with an absorption maximum at 505 nm. Beer's law is valid over the concentration range of 0.5 - 6.5 microg ml(-1) of thallium(III). The molar absorptivity and Sandell's sensitivity of the color system are 2.14 x 10(4) l mol(-1) cm(-1) and 0.0095 microg cm(-2), respectively. The optimum reaction conditions and other analytical parameters were evaluated. The tolerance limit of the method towards various ions usually associated with thallium has been studied. The proposed method has been successfully applied to the analysis of thallium in alloys, minerals, standard reference material, water, and urine samples.

Alloys↗

Determination of mercury in biological tissues by graphite-furnace atomic absorption spectrometry with an in-situ concentration technique.

A method has been described for the determination of total mercury by graphite-furnace atomic absorption spectrometry (GFAAS) using an in-situ concentration technique with a Pd-Zr coating and a chemical modifier. The characteristic mass, which gives an integrated absorbance of 0.0044 s, was found to be 42 pg and an absolute detection limit (3sigma) of 33 pg was obtained with the proposed modifier. The total mercury values in standard reference materials, including Mussel (GBW08571), Bovine liver (GBW08306), Peach leaf (GBW08501) and Tea leaf (GBW080001), were determined using the proposed method, and the results were consistent with reference values. The method had been successfully applied to the determination of mercury in biological tissue samples with a recovery range of 94-105%.

Animals↗

Simultaneous determination of gallium and indium with 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol in cationic micellar medium using derivative spectrophotometry.

A new derivative spectrophotometric method for rapid and selective trace analysis of Ga3+ and In3+ and for their simultaneous determination using 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol in a cationic micellar medium is reported. Molar absorptivity and Sandell's sensitivity of 1:1 Ga+ and In3+ complexes at their lambda(max) 553 nm and 558 nm are: 7.22 x 10(4) l mol(-1) cm(-1) and 5.85 x 10(4) l mol(-1) cm(-1), and 0.96 ng cm(-2) and 1.96 ng cm(-2), respectively. Linearity is observed in the concentration range 0.023-0.700 microg ml(-1) for gallium and 0.076-1.52 microg ml(-1) for indium; IUPAC detection limit is 0.012 and 0.035 ng ml(-1), respectively. These metal ions interfere with the determination of each other. However, 0.07-0.70 microg ml(-1) Ga3+ and 0.115-1.150 microg ml(-1) In3+ could be determined simultaneously when present together by the derivative method without any prior separation. The proposed procedures have been successfully applied for the individual and simultaneous determination of gallium and indium in synthetic binary mixtures, standard reference materials and environmental samples.

Azo Compounds↗

Loudness adaptation: real or illusory.

There is general agreement that, except at low sensation levels, the loudness of a continuous pure tone does not appear to change with time. By contrast, using a simultaneous dichotic loudness balancing procedure (SDLB) a substantial fall in loudness can be revealed. As a result it has been held that the latter does not represent a loudness loss in real terms but is a manifestation of some form of central binaural interaction possibly involving localization mechanisms. In two studies reported here, one involving a loudness scaling procedure using as a standard a reference tone of 65 dB (HL) and the other a loudness doubling procedure, a loudness loss has been clearly demonstrated following sustained stimulation with a continuous 60-dB (HL), 1000-Hz tone applied monaurally. The progression of the loudness loss with time follows a similar course to that found with the SDLB procedure. In consequence it is concluded that both represent a loudness loss in real terms and that inability to detect a change in loudness over time of a continuous tone results from the absence of any reference standard by which it can be judged.

Acoustic Stimulation↗

Standards for autoantibody testing; addressing future needs for autoimmune disease and cancer diagnosis.

It is widely held that determination of increased levels of serum autoantibodies can play an important role in the diagnosis and, in some cases, management of many autoimmune diseases. It is also widely held that such testing is fraught with problems relating to accuracy and detection of clinically relevant autoantibodies. Attempts to standardize autoantibody testing by making available standard reference sera, have helped to some degree to improve the testing. Nevertheless, problems still exist and may be causing the delayed diagnosis of severely debilitating and sometimes fatal autoimmune diseases. This paper discusses current practice, as well as, possible new approaches intended to make more robust, autoantibody determinations in all fields of medicine.

Autoantibodies↗

The diagnostic value of magnetic resonance imaging in subacromial impingement syndrome.

The aim of this study was to assess the diagnostic ability of magnetic resonance imaging (MRI) in subacromial impingement syndrome (SIS), using a physiological standard of reference. MRI of the rotator cuff (RC) and subacromial injection test (SIT), a reference standard for SIS diagnosis, were performed in 125 painful shoulders. MRI diagnostic accuracies were determined using a 2 x 2 table and the percentage values of SIS diagnosis in patients with the three Zlatkin MRI stages were determined. Shoulder function was evaluated using the Constant Scale, and results were compared for stages. The sensitivity, specificity, accuracy, positive and negative predictive values of MRI for SIS diagnosis were 98.85%, 36.84%, 80%, 78.18% and 93.33% respectively. Of the 32 patients with Zlatkin stage 1 changes in MRI, 20 (62%) had SIT approved SIS diagnosis, while 47 (79%) of the 59 patients with Zlatkin 2 and all of the 19 (100%) patients with Zlatkin 3 changes were diagnosed with SIS by SIT. Mean Constant scores were 78.04 +/- 18.3, 65.0 +/- 19.9 and 54.52 +/- 20.7 in patients with Zlatkin stages 1, 2 and 3, respectively (p < 0.05). The MRI of RC did not prove to be an excellent tool for SIT based SIS diagnosis, with its low specificity. However, the technique can give important clues, as its sensitivity and negative predictive values are high.

Adolescent↗

Neutron activation analysis of biological materials by the monostandard method.

Instrumental neutron activation analysis by the monostandard method has been applied to the analyses of biological NBS standard reference materials; 1571 Orchard Leaves and 1577 Bovine Liver. Aluminum foils containing 0.100% gold or 2.00% cobalt were used as the monostandards. The gamma-ray spectral data were recorded on punched paper tape and were analyzed by a computer assisted data processing. The following 25 elements were determined: Al, Ca, Cl Cu, Mg, Mn, V (by short period irradiation), As, Ba, Br, Co, Cr, Cs, Eu, Fe, Hg, K, La, Na, Rb, Sb, Sc, Se, Sm and Zn (by long period irradiation). The results were compared with the certified values by NBS and the reported values in literatures to prove the reliability and accuracy of the monostandard method.

Activation Analysis↗

Determination of selenium in meat products by hydride generation atomic absorption spectrophotometry.

Wet digestion using a mixture of nitric, sulfuric, and perchloric acids and an aluminum block digester effectively and rapidly decomposed meat samples for selenium determination by hydride generation atomic absorption spectrophotometry. Digestion did not require constant attention by an operator. Selenium recoveries (range, 94-105%) from National Institute of Standards and Technology standard reference materials and spiked samples were used to validate method accuracy. Coefficients of variation (CVs) of repeatability of in-house reference materials used for precision study were 6.4 and 5.6%, respectively, for seafood mix and mutton liver. Selenium levels in meat products from Brisbane markets varied widely: 0.042-0.142, 0.081-0.42, and 0.050-0.198 microgram/g (wet weight) respectively, for beef, chicken, and pork. Overall, selenium levels in manufactured meat ranged from 0.041 to 0.189 microgram/g. The levels of selenium found in this study were generally lower than those reported in Finland but comparable with those reported in some parts of the United States.

Animals↗