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At least 1,153 records · Page 64Linked to original sources
PubMed1996

Comparative evaluation of an automated ribotyping system versus pulsed-field gel electrophoresis for epidemiological typing of clinical isolates of Escherichia coli and Pseudomonas aeruginosa from patients with recurrent gram-negative bacteremia.

Ribotyping and macrorestriction analysis of chromosomal DNA using pulsed-field gel electrophoresis (PFGE) are among the more useful molecular epidemiologic typing methods. Because these techniques are labor intensive, automation of one or more steps may allow clinical laboratories to apply molecular typing methods. We compared the recently developed automated ribotyping system, the RiboPrinter Microbial Characterization System (DuPont), with PFGE as a means of typing clinical isolates of E. coli and P. aeruginosa. A total of 22 E. coli and 24 P. aeruginosa were typed by both PFGE and the RiboPrinter. When compared with PFGE typing of E. coli and P. aeruginosa, the RiboPrinter was less sensitive in identifying different strains, particularly among the isolates of P. aeruginosa. The RiboPrinter was completely automated and allowed up to 32 isolates to be typed within an 8-hour period. The pattern of results obtained in this study suggests that a heirarchical approach to molecular typing using the RiboPrinter Microbial Characterization System plus PFGE might be feasible. The RiboPrinter Microbial Characterization System promises to be a very useful addition to the expanding molecular typing armamentarium.

Bacteremia↗
PubMed1987

Advanced technology and its impact on the clinical laboratory.

The analytical capabilities of the clinical laboratory have continued to expand and improve as a result of technical developments and advancements made in a wide spectrum of allied disciplines. As a consequence, the clinical laboratory has evolved from the small, manual operations of yesterday to the large, central organizations of today. Technology continues to have an impact on the laboratory, especially with the advent of the fully automated analytical systems that are having such a profound effect on how the laboratory is staffed, equipped, organized, and operated. With the accelerating rate at which new developments are occurring, it is safe to assume that dramatic changes will continue to occur in this area. Consequently, it will become increasingly important for clinical laboratorians to be aware of these new developments, to understand them, and to anticipate how they will be assimilated and integrated into the overall health care system. To quote from a recent Bell Atlantic advertisement, "The genius of the future lies not in technology alone, but in the ability to manage it."

Chemistry, Clinical↗
PubMed1998

[New technology of automated blood cell differential counting].

Examination of stained peripheral blood films for evaluation of erythrocyte and platelet morphology and leukocyte differential count provides considerable data for diagnosis of various diseases including hematologic disorders. Since 1980, the use of an automated leukocyte differentiation analyzer has become common in large clinical laboratories. We surveyed the principles, performance, and characteristics of the newest automated leukocyte differential instruments, GEN.S system (Coulter), SE-9000/RAM-1 (Toa), CELL-DYN 4000 (Dainabot), ADVIA120 (Bayer-Sankyo), VEGA RETIC LC-141 (Horiba), and MICROX HEG-120N (Omron). We also compared the differentiated function of GEN.S system and CELL-DYN 4000. Automated leukocyte differentiation using two instruments correlated well with a reference method for leukocyte differential counting proposed by the National Committee for Clinical Laboratory Standards (NCCLS) H20-T (eye-count differential). The sensitivities of the suspect flag in GEN.S system and CELL-DYN 4000 were 79.3% and 83.5%, and the specificities of the two instruments were 100% and 94.9%, respectively.

Humans↗
PubMed2005

Application of the BioRobot EZ1 in a forensic laboratory.

In a forensic laboratory, the routine application of an automated DNA extraction and purification robot has to fulfil several conditions, like producing reproducible DNA's of sufficient quantity and quality from all the different forensic biological stains relevant to various carrier materials. In this study, the suitability of the BioRobot EZ1 system from QIAGEN (Hilden, Germany), which offers fully automated extraction and purification of nucleic acids using magnetic bead technology, was tested. In summary, the DNA's obtained from the BioRobot EZ1 for different forensic relevant biological materials showed a quantity and quality comparable to those of the forensic standard protocols normally used in our laboratory. The system saves time, because there is no need of any further purification or concentration step after the automated DNA extraction. It can also be used as a replacement for time consuming organic extractions. A disadvantage of the system was the unsteady quality of the chemical regencies used by the robot. Nineteen different lots were tested with a self designed test system.

DNA↗
PubMed2006

Capillary electrophoresis and the clinical laboratory.

Over the past 15 years, CE as an analytical tool has shown great promise in replacing many conventional clinical laboratory methods, such as electrophoresis and HPLC. CE's appeal was that it was fast, used very small amounts of sample and reagents, was extremely versatile, and was able to separate large and small analytes, whether neutral or charged. Because of this versatility, numerous methods have been developed for analytes that are of clinical interest. Other than molecular diagnostic and forensic laboratories CE has not been able to make a major impact in the United States. In contrast, in Europe and Japan an increasing number of clinical laboratories are using CE. Now that automated multicapillary instruments are commercially available along with cost-effective test kits, CE may yet be accepted as an instrument that will be routinely used in the clinical laboratories. This review will focus on areas where CE has the potential to have the greatest impact on the clinical laboratory. These include analyses of proteins found in serum and urine, hemoglobin (A1c and variants), carbohydrate-deficient transferrin, forensic and therapeutic drug screening, and molecular diagnostics.

Blood Proteins↗
PubMed1977

Automated blood grouping of patients and blood donors using Groupamatic 360 S and a separate mini-computer. System description and evaluation.

An automated blood-grouping system for all kinds of blood samples is described. The system is built upon Groupamatic 360 S and a separate mini-computer. Both the laboratory and the clerical work is automated. Blood group replies are given on coloured punch cards in the Scandinavian blood group colours. Lists of different groups of samples can be compiled. After 1 year of routine use, the refusal rate is about 5%. The system is reliable and rapid, and permits saving, both of manpower and reagents.

Blood Group Antigens↗
PubMed1982

Using the proposed NCCLS protocol for evaluation of automated instruments.

We compared four instruments (Sequential Multiple Analyzer Computer (SMAC), AutoAnalyzer II (AAII), StatLyte, and PVA-4) for the determination of Na, K, Cl and CO2 using a protocol (PSEP-1) of the National Committee for Clinical Laboratory Standards (NCCLS) for evaluation of precision and accuracy of automated instruments. The protocol was applied concurrently to all four instruments. NCCLS has revised this protocol into three separate protocols by which the manufacturer can make performance claims. Protocols specifically aimed at the user are now under development. Although PSEP-1 is no longer in use, many parts of it have been incorporated into PSEP-2, PSEP-3 and PSEP-4. This article will describe the adaptation of the protocol to evaluate four automated instruments in our laboratory.

Autoanalysis↗
PubMed1998

Rapid, efficient method for multiplex amplification from filter paper.

Guthrie cards derived from the New York State Newborn Screening Program were utilized to develop a rapid, economical method for amplifying multiple genes to detect mutations that impact public health. These specimens are untraceable to the donor because identifiers are removed and discarded; therefore, these pilot studies were carried out anonymously. The sample preparation requires minimal manipulation, is amenable to automation, and is useful in laboratories which routinely process large numbers of samples, such as those in typical newborn screening laboratories. Multiple gene fragments may be amplified from a 1 mm punch which contains less than 1 microl of whole blood. The blood spots used in these studies contain sufficient material for up to 25 amplification reactions which multiplex at least four different gene fragments each. Since sufficient material remains on the card after the routine testing is complete, this simple assay can greatly expand the efficacy of current newborn screening programs by permitting DNA diagnosis of some disorders when indicated, particularly those in which genotype-phenotype correlations are useful. In addition to newborn screening specimens, this method is also applicable to whole blood from adults after phlebotomy and from lymphoblastoid cell lines. Use of filter paper for DNA analysis is particularly useful for shipped specimens or for population studies whose subjects are refractory to phlebotomy.

Alleles↗
PubMed2005

Evaluation of single-tube osmotic fragility as a screening test for thalassemia.

A single-tube osmotic fragility test has been proposed for thalassemia screening with a range of different concentrations of saline having been employed. We have compared the sensitivity and specificity of 0.32%, 0.34%, and 0.36% buffered saline, and on the basis of our findings, recommend the use of 0.36% saline. This gave definitely positive or equivocal results in 81 of 85 patients with beta thalassemia trait and in 4 of 4 with alpha(0) thalassemia trait. There were 14% false positive results in hematologically normal patients and 81% of the samples from patients with various variant hemoglobins gave positive results. The sensitivity was 95% and specificity 86%. The single-tube osmotic fragility test is potentially useful in under-resourced laboratories although it cannot replace automated red cell indices using electronic counters.

False Positive Reactions↗
PubMed2003

N-methyl-D-glucamine improves the laser-induced fluorescence capillary electrophoresis performance in the total plasma thiols measurement.

We describe an ultrarapid capillary electrophoresis with laser-induced fluorescence (CE-LIF) method for total plasma thiols measurement. Reduced thiols by 10% tri-n-butylphosphine (TBP) were derivatized in 10 min at room temperature with 5-iodoacetamidofluorescein (5-IAF) as fluorescent reagent. We show that CE-LIF allows a baseline separation of total plasma cysteinylglycine, homocysteine, cysteine, and glutathione in less than 5 min when N-methyl-D-glucamine in run buffer was added. CE was compared with high-performance liquid chromatography (HPLC) with fluorescence detection. The Bland-Altman test and Passing-Bablok regression demonstrates that the results obtained by CE-LIF and by HPLC are highly comparable. The simplified procedure of sample preparation, the short incubation and fast separation times, the high specificity, sensitivity and reproducibility, and the lower cost of analysis suggest that our proposed method can be considered valuable for the automation analysis in a routine laboratory.

Adult↗
PubMed2002

Prediction of vapour pressures for halogenated diphenyl ether congeners from molecular descriptors.

BACKGROUND, AIM AND SCOPE: Polychlorinated diphenyl ethers (PCDE) and polybrominated diphenyl ethers (PBDE) have both been identified as environmental contaminants. The physical properties are important in determining the distribution and fate of organic contaminants in the environment. The purpose of the present investigation was to characterise halogenated diphenyl ethers using computationally derived descriptors, and to develop calibration models for the vapour pressure from published experimental data. METHODS: Experimental data for vapour pressures were obtained from the literature. The chemical structure of each PCDE and PBDE congener was optimised prior to descriptor generation. The data analysis was performed using principal component analysis (PCA) and partial least squares regression (PLSR). The calibration models were validated with external test sets. RESULTS AND DISCUSSION: All congeners of PCDEs and PBDEs were characterised by 795 molecular descriptors and two principal components could account for about two thirds of the variance within each group. Bilinear calibration models were developed that could explain 99.4% of the variance in the external validation test sets. Vapour pressures were subsequently predicted for all congeners that were adequately described by these calibration models. The type and number of halogen atoms in the molecule were the main factors influencing the vapour pressures of halogen substituted diphenyl ethers, but the variations in substitution pattern was also shown to be a significant factor. CONCLUSIONS: The molecular descriptor patterns of halogenated aromatic compounds such as diphenyl ethers can be described and interpreted using principal component analysis (PCA). The major sources of variation in the descriptor spaces for PCDEs and PBDEs are the same as those contributing to the differences in vapour pressure, similar to what has previously been reported for the PCBs. The bilinear calibration models for vapour pressure presented here, has a standard error of prediction that is lower than what is reported as the experimental uncertainty or observed as deviations between experimental investigations. The estimated prediction errors are expected to be within the reported boundaries when the models are applied to new objects within the same molecular descriptor space, and model predictions can hence extend the current database of experimental values. RECOMMENDATIONS AND OUTLOOK: The results from this investigation and others show that the establishment of quantitative structure-property relationships (QSPR) is a viable approach to estimate physical properties for halogenated diphenyl ethers. It is easy to foresee an increased need for using QSPR estimation methods in the future, for evaluation of the environmental fate for organic pollutants. Despite method developments and automation, it is unlikely that laboratory determinations can cope with the pace that new pollutants are identified.

Calibration↗
PubMed2006

Serum insulin-like growth factor-I measurement in the follow-up of treated acromegaly: comparison of four immunoassays.

BACKGROUND: Measurement of serum insulin-like growth factor I (IGF-I) is currently used for the diagnosis and for monitoring treated acromegaly. In this work, we have studied the performances of four IGF-1 immunoassays in the follow-up of acromegaly. METHODS: The study was carried out on 52 sera from 40 patients with treated acromegaly. Serum IGF-I was measured with radioimmunoassays from Immunotech, DiaSorin and Schering Laboratories and by the chemiluminescent automated immunoassay (Advantage) from Nichols. Normal age- and gender-matched subjects constituted the reference population for two assays. RESULTS: Deming regression and Bland-Altman analyses showed a high correlation with the four methods tested. The use of restricted number of age-adjusted controls with the Immunotech assay, the DiaSorin assay and the Schering assay induced discordance with the Nichols assay in the interpretation of results in treated acromegaly. When normal ranges of the Immunotech assay and the DiaSorin assay were defined from large number of controls, the frequency of normal IGF-I became similar as that obtained with the Nichols assay. CONCLUSION: The four immunoassays display suitable analytical performance for serum IGF-I measurement in the follow-up of acromegaly. For correct interpretation, IGF-I normal ranges should be defined in a large number of age-adjusted healthy subjects.

Acromegaly↗
PubMed1999

A short echo proton magnetic resonance spectroscopy study of the left mesial-temporal lobe in first-onset schizophrenic patients.

BACKGROUND: Past 1H magnetic resonance spectroscopy (MRS) studies of the temporal lobe in schizophrenic patients have shown decreased levels of N-acetylaspartate (NAA) suggesting reduced neuronal density in this region. However, the measured volumes have been large and included contributions from mostly white matter. METHODS: Short echo 1H MRS was used to measure levels of NAA and other metabolites (i.e., glutamate and glutamine) from a 6 cm3 volume in the left mesial-temporal lobe of 11 first-episode schizophrenic patients and 11 healthy control subjects of comparable age, gender, handedness, education, and parental education levels. Spectra were quantified without operator interaction using automated software developed in our laboratory. Metabolite levels were normalized to the internal water concentration of each volume studied. Images were also obtained to determine temporal lobe gray and white matter volumes. RESULTS: No significant differences were found between levels of NAA or other metabolites, or gray and white matter volumes, in first-episode schizophrenic patients and comparison subjects. CONCLUSIONS: Since the volume studied was small compared to previous studies and contained mostly gray matter, this result suggests consequential NAA decreases may be restricted to regions of white matter.

Adolescent↗
PubMedDate not supplied

Three manual immunoassays for the rapid detection of hepatitis B surface antigen.

We have evaluated 3 commercial test kits for the detection of hepatitis B surface antigen (HBsAg) which do not require elaborate instrumentation and are relatively inexpensive. These included the Immunocomb-II (Orgenics, Israel), Austragen latex agglutination (Span, India) and Daina Screen (Abbott, USA) tests. A panel of 98 sera tested by IMx HBsAg assay (Abbott, USA) were used to evaluate the tests. Sensitivities of 96.6%, 86.2% and 96.6%, and specificities of 95.7%, 94.2% and 100%, respectively, were obtained. We therefore recommend the Immunocomb-II or Daina Screen tests for HBsAg detection in small laboratories where facilities to conduct automated enzyme-linked immunosorbent assays are not available.

Evaluation Studies as Topic↗
PubMed2004

The European Astronaut Centre prepares for International Space Station operations.

The European Space Agency (ESA) contribution to the International Space Station (ISS) goes much beyond the delivery of hardware like the Columbus Laboratory, its payloads and the Automated Transfer Vehicles. ESA Astronauts will be members of the ISS crew. ESA, according to its commitments as ISS international partner, will be responsible to provide training on its elements and payloads to all ISS crewmembers and medical support for ESA astronauts. The European Astronaut Centre (EAC) in Cologne has developed over more than a decade into the centre of expertise for manned space activities within ESA by contributing to a number of important co-operative spaceflight missions. This role will be significantly extended for ISS manned operations. Apart from its support to ESA astronauts and their onboard operations, EAC will have a key role in training all ISS astronauts on ESA elements and payloads. The medical support of ISS crew, in particular of ESA astronauts has already started. This paper provides an overview on status and further plans in building up this homebase function for ESA astronauts and on the preparation towards Training Readiness for ISS crew training at EAC, Cologne. Copyright 2001 by the European Space Agency. Published by the American Institute of Aeronautics and Astronautics, Inc., with permission. Released to IAF/IAA/AIAA to publish in all forms.

Aerospace Medicine↗
PubMed1999

High-throughput extraction, amplification, and detection (HEAD) of HCV-RNA in individual blood donations.

BACKGROUND: High-throughput nucleic acid amplification techniques (NATs) are required for the detection of viral genomes in individual blood donations and might be helpful in any virological laboratory. OBJECTIVE: To develop and automate a method for the detection of hepatitis C virus RNA in individual blood donations, compatible with the time schedule of routine blood bank screening an product release. STUDY DESIGN: The viral RNA was isolated with the use of target specific capture oligonucleotides and magnetic beads. This extraction method was combined with reverse transcription/amplification (RT/PCR) and fluorescence detection. We adapted our method on a pipetting robot and pipetted all steps in a single room. When the pipetting was completed, microtiter plates were heat-sealed with foils and placed into a thermocycler. Positive reactions were detected with a fluorescent dye in a second room. Aerosols were avoided with programmed slow pipetting steps and with a special device constructed for the removal of the used disposable tips. During a 7 month period, we used this method in routine testing of individual donations prior to the release of all blood components. RESULTS: The total number of 11,700 individual donations including platelet concentrates were analysed. We tested up to 192 specimens in one run within 7 h. The frequency of cross-contamination using the automated procedure was 0.1%. Five specimens have been found repeatedly reactive for HCV-RNA, four of these were anti-HCV positive, one sample from a repeat donor was negative in anti-HCV assays. A seroconversion was detectable at his next presentation, 6 months later. CONCLUSION: In this pilot study, we demonstrate that automated HCV-RT-PCR testing is practicable for individual donations in high-throughput. Additionally, the described PCR approach could easily be adapted to the detection of other viral genomes by the use of specific primers.

Automation↗