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At least 1,063 records · Page 59Linked to original sources
PubMed2005

Quantification of red blood cell fragmentation by the automated hematology analyzer XE-2100 in patients with living donor liver transplantation.

The fragmented red cell (FRC) is a useful index for diagnosing and determining the severity of thrombotic thrombocytopenic purpura (TTP), thrombotic microangiopathy (TMA) and other similar conditions, as it is found in peripheral blood in patients with these diseases. The FRC expression rate has conventionally been determined by manual methods using smear samples. However, it is difficult to attain accurate quantification by such methods as they are time consuming and prone to a great margin of error. With cases of living donor liver transplantation, the current study examined the possibility of using a multi-parameter automated hematology analyzer, the XE-2100 (Sysmex Corporation) for FRC quantification. While there was a notable correlation between the manual and automated measurements, the manual measurement resulted in higher values. This suggested remarkable variations in judgment by individuals. The FRC values had a significant correlation with the reticulocyte count, red blood cell distribution width (RDW), fibrin/fibrinogen degradation products (P-FDP) and lactate dehydrogenase (LDH) among the test parameters, and this finding was consistent with the clinical progression in patients. The automated method can offer precise measurements in a short time without inter-observer differences, meeting the requirement for standardization. The determination of FRC count (%) by the XE-2100 that enables early diagnoses and monitoring of TTP or TMA will be useful in the clinical field.

Adolescent↗
PubMed2006

Automated red cell collection--quality and value.

Automated red cell collection was initially used largely for therapeutic purposes. New technology has rendered the procedure safer for donors and easier for machine operators. Optimal additive solution can be automatically added and the red cells filtered to provide a leucodepleted product. Two units of red cells may be collected during a single procedure from individuals who have a high enough red cell mass, whilst a single unit of red cells plus platelets or plasma can be collected from smaller donors. In vitro studies suggested that red cells collected by automated methods would be of better quality than those collected by gravity. This was not confirmed in vivo, but red cells collected by automated methods have the major advantage of consistency in terms of haemoglobin content, volume and haematocrit, compared with red cells collected by gravity. This standardised product is of particular value for transfusion dependent patients as the patient's haemoglobin can be maintained within narrow limits. The use of a double dose red cell product for transfusion to a single patient also confers benefit in terms of reduction in donor exposure.

Automation↗
PubMed1986

Automation in blood banking.

During the past decade, automation took a major place in blood transfusion laboratories. Now many centers have purchased automated equipment and subsequently undertaken the computerization of their activities. The aim of this paper is to review the state of the art in the field of automation and to try to estimate future tendencies.

Automation↗
PubMed2005

von Willebrand factor ristocetin cofactor (VWF:RCo) assay: implementation on an automated coagulometer (ACL).

BACKGROUND: VWF:RCo assay is the standard and widely used laboratory test for von Willebrand disease (VWD) diagnosis. It is hampered by high intra- and inter-assay imprecision and is time consuming. Automation may improve the assay performance and allow its routine application. OBJECTIVE: Automation of VWF:RCo on the ACL 7000 coagulometer (Instrumentation Laboratory, Milan, Italy) and its evaluation in VWD diagnosis. METHODS AND MATERIALS: Method performance determination: precision, detection limit (DL), interferences, dose-response curve. Method comparison: analysis of 105 plasma samples from normal subjects (50), VWD type 1 (24), VWD type 2 (24) and VWD type 3 (7) with ACL VWF:RCo and comparison with the reference aggregometric (AGM) method. RESULTS: ACL VWF:RCo: CVs around 10% vs. 19% of AGM method; DL: 0.08 U mL(-1); potential interferences from bilirubin, triglycerides and hemoglobin, avoided by suitable plasma dilution; high correlation with AGM VWF:RCo (Deming regression Y =-0.0277 + 1.0519X) either in normal or VWD plasmas. In VWD types 1 and 2, the VWF:RCo/VWF:Ag ratios are >0.6 or <0.6, respectively, when calculated with both AGM and ACL VWF:RCo values. CONCLUSIONS: The automated VWF:RCo on the ACL 7000 coagulometer shows precision improvement and high correlation with the reference AGM method. The test allows the diagnosis of both quantitative (VWD types 1 and 3) and qualitative (VWD type 2) forms of the disease. These results and the assay feasibility make it a suitable and reliable test for the routine diagnosis of VWD.

Automation↗
PubMed2001

Automated single-cell sorting system based on optical trapping.

We provide a basis for automated single-cell sorting based on optical trapping and manipulation using human peripheral blood as a model system. A counterpropagating dual-beam optical-trapping configuration is shown theoretically and experimentally to be preferred due to a greater ability to manipulate cells in three dimensions. Theoretical analysis performed by simulating the propagation of rays through the region containing an erythrocyte (red blood cell) divided into numerous elements confirms experimental results showing that a trapped erythrocyte orients with its longest axis in the direction of propagation of the beam. The single-cell sorting system includes an image-processing system using thresholding, background subtraction, and edge-enhancement algorithms, which allows for the identification of single cells. Erythrocytes have been identified and manipulated into designated volumes using the automated dual-beam trap. Potential applications of automated single-cell sorting, including the incorporation of molecular biology techniques, are discussed.

Automation↗
PubMed2001

Automated detection of lung nodules in CT scans: preliminary results.

We have developed a fully automated computerized method for the detection of lung nodules in helical computed tomography (CT) scans of the thorax. This method is based on two-dimensional and three-dimensional analyses of the image data acquired during diagnostic CT scans. Lung segmentation proceeds on a section-by-section basis to construct a segmented lung volume within which further analysis is performed. Multiple gray-level thresholds are applied to the segmented lung volume to create a series of thresholded lung volumes. An 18-point connectivity scheme is used to identify contiguous three-dimensional structures within each thresholded lung volume, and those structures that satisfy a volume criterion are selected as initial lung nodule candidates. Morphological and gray-level features are computed for each nodule candidate. After a rule-based approach is applied to greatly reduce the number of nodule candidates that corresponds to nonnodules, the features of remaining candidates are merged through linear discriminant analysis. The automated method was applied to a database of 43 diagnostic thoracic CT scans. Receiver operating characteristic (ROC) analysis was used to evaluate the ability of the classifier to differentiate nodule candidates that correspond to actual nodules from false-positive candidates. The area under the ROC curve for this categorization task attained a value of 0.90 during leave-one-out-by-case evaluation. The automated method yielded an overall nodule detection sensitivity of 70% with an average of 1.5 false-positive detections per section when applied to the complete 43-case database. A corresponding nodule detection sensitivity of 89% with an average of 1.3 false-positive detections per section was achieved with a subset of 20 cases that contained only one or two nodules per case.

Adult↗
PubMed2006

Automated detection of lung nodules in CT scans: false-positive reduction with the radial-gradient index.

We present a number of approaches based on the radial gradient index (RGI) to achieve false-positive reduction in automated CT lung nodule detection. A database of 38 cases was used that contained a total of 82 lung nodules. For each CT section, a complementary image known as an "RGI map" was constructed to enhance regions of high circularity and thus improve the contrast between nodules and normal anatomy. Thresholds on three RGI parameters were varied to construct RGI filters that sensitively eliminated false-positive structures. In a consistency approach, RGI filtering eliminated 36% of the false-positive structures detected by the automated method without the loss of any true positives. Use of an RGI filter prior to a linear discriminant classifier yielded notable improvements in performance, with the false-positive rate at a sensitivity of 70% being reduced from 0.5 to 0.28 per section. Finally, the performance of the linear discriminant classifier was evaluated with RGI-based features. RGI-based features achieved a substantial improvement in overall performance, with a 94.8% reduction in the false-positive rate at a fixed sensitivity of 70%. These results demonstrate the potential role of RGI analysis in an automated lung nodule detection method.

Algorithms↗
PubMed1998

Automated lung segmentation in digital lateral chest radiographs.

We are developing a fully automated computerized scheme for segmenting the lung fields in digital lateral chest radiographs. Existing computer-aided diagnostic (CAD) schemes and automated lung segmentation methods have focused exclusively on the posteroanterior view, despite the diagnostic importance of the lateral view. Information from the lateral radiograph is routinely incorporated by radiologists in their decision-making process, and thus computer analysis of lateral images may potentially add another dimension to current CAD schemes. Automated analysis of the lung fields in lateral images will necessarily require accurate segmentation. Our scheme employs an initial procedure to eliminate external and subcutaneous pixels. Global gray-level histogram analysis then allows for the identification of a range of gray-level thresholds. An iterative gray-level thresholding scheme is implemented using this range of thresholds to construct a series of binary images in which contiguous regions are identified and geometrically analyzed. Regions determined to be outside the lungs are prevented from contributing to binary images at later iterations. Adaptive local gray-level thresholding is applied along the initial contour that results from the global thresholding procedure to extend the contour closer to the true lung borders. This local thresholding method uses regions of interest of various dimensions, depending on the enclosed anatomy. Smoothing and polynomial curve fitting complete the segmentation. A database of 100 normal and 100 abnormal lateral images was analyzed. Quantitative comparison of computer-segmented lung regions with lung regions manually delineated by two radiologists indicated that 83% and 84% of normal and abnormal images, respectively, displayed segmentation contours within three standard deviations of the mean inter-radiologist contour degree-of-overlap value.

Automation↗
PubMed2003

Automated enumeration of groups of marine picoplankton after fluorescence in situ hybridization.

We describe here an automated system for the counting of multiple samples of double-stained microbial cells on sections of membrane filters. The application integrates an epifluorescence microscope equipped with motorized z-axis drive, shutters, and filter wheels with a scanning stage, a digital camera, and image analysis software. The relative abundances of specific microbial taxa are quantified in samples of marine picoplankton, as detected by fluorescence in situ hybridization (FISH) and catalyzed reporter deposition. Pairs of microscopic images are automatically acquired from numerous positions at two wavelengths, and microbial cells with both general DNA and FISH staining are counted after object edge detection and signal-to-background ratio thresholding. Microscopic fields that are inappropriate for cell counting are automatically excluded prior to measurements. Two nested walk paths guide the device across a series of triangular preparations until a user-defined number of total cells has been analyzed per sample. A backup autofocusing routine at incident light allows automated refocusing between individual samples and can reestablish the focal plane after fatal focusing errors at epifluorescence illumination. The system was calibrated to produce relative abundances of FISH-stained cells in North Sea samples that were comparable to results obtained by manual evaluation. Up to 28 preparations could be analyzed within 4 h without operator interference. The device was subsequently applied for the counting of different microbial populations in incubation series of North Sea waters. Automated digital microscopy greatly facilitates the processing of numerous FISH-stained samples and might thus open new perspectives for bacterioplankton population ecology.

Automation↗
PubMed2004

Automated purification and suspension array detection of 16S rRNA from soil and sediment extracts by using tunable surface microparticles.

Autonomous, field-deployable molecular detection systems require seamless integration of complex biochemical solutions and physical or mechanical processing steps. In an attempt to simplify the fluidic requirements for integrated biodetection systems, we used tunable surface microparticles both as an rRNA affinity purification resin in a renewable microcolumn sample preparation system and as the sensor surface in a flow cytometer detector. The tunable surface detection limits in both low- and high-salt buffers were 1 ng of total RNA ( approximately 10(4) cell equivalents) in 15-min test tube hybridizations and 10 ng of total RNA ( approximately 10(5) cell equivalents) in hybridizations with the automated system (30-s contact time). RNA fragmentation was essential for achieving tunable surface suspension array specificity. Chaperone probes reduced but did not completely eliminate cross-hybridization, even with probes sharing <50% identity to target sequences. Nonpurified environmental extracts did not irreparably affect our ability to classify color-coded microparticles, but residual environmental constituents significantly quenched the Alexa-532 reporter fluor. Modulating surface charge did not influence the interaction of soluble environmental contaminants with conjugated beads. The automated system greatly reduced the effects of fluorescence quenching, especially in the soil background. The automated system was as efficacious as manual methods for simultaneous sample purification, hybridization, and washing prior to flow cytometry detection. The implications of unexpected target cross-hybridization and fluorescence quenching are discussed relative to the design and implementation of an integrated microbial monitoring system.

Automation↗
PubMed1979

Clinical laboratory evaluation of automated microbial detection/identification system in analysis of clinical urine specimens.

More than 4,000 clinical urine specimens were evaluated with an automated microbial detection/identification system compared to a standarized manual analysis and the routine modalities used in five peer-group laboratories. The comparison indicates that the automated system recognizes the nine groups of significant microorganisms in urinary tract infections in hospitalized patients with the same efficiency as a standarized manual method. The automated system's ability to enumerate the bacterial populations in the original clinical specimen attained a high degree of accuracy.

Automation↗
PubMed1996

Evaluation of automated COBAS AMPLICOR PCR system for detection of several infectious agents and its impact on laboratory management.

We evaluated the COBAS AMPLICOR (CA) PCR system (Roche Diagnostic Systems) designed for automated PCR amplification and detection of nucleic acids from infectious agents in clinical samples. The Roche AMPLICOR microwell plate (MWP) PCR was the reference method. CA amplifies target nucleic acid, captures the biotinylated amplification products by using magnetic particles coated with specific oligonucleotide probes, and detects the bound products colorimetrically. For Mycobacterium tuberculosis, the correlation of the results of CA tests with those of MWP tests was 100% with 230 samples, including 20 culture-positive samples. For hepatitis C virus, the correlation was 100% with 214 samples, including 60 positive samples. MultiPlex CA analysis of 199 cervical specimens for Chlamydia trachomatis, Neisseria gonorrhoeae, and the internal control gave 100% concordance. These samples included 19 C. trachomatis and 3 N. gonorrhoeae culture-positive samples. Overall, the agreement between PCR methods for all 842 comparisons was 100%. Compared with culture, the sensitivities of the assays for C. trachomatis and M tuberculosis were > or = 95%. After spiking alternating amplification tubes in the CA system with 10(14) copies of the Chlamydia amplicon per ml, we were unable to demonstrate any carryover cross-contamination of negative samples. Using the criteria of the College of American Pathologists workload recording method, we found that the total hands-on time to produce CA PCR results was 4.4, 7.9, and 3.3 min for M. tuberculosis, hepatis C virus, and the MultiPlexed assay for chlamydia plus gonorrhea and an internal control, respectively. The CA system brings true PCR automation to laboratories. In addition to the accuracy of automated results, the CA system provides labor savings, provides containment of the amplification and detection components of PCR, and supports both MultiPlex amplification and sequential algorithm (ReFlex) detection of analytes.

Automation↗
PubMed2000

Distinguishing species of the Burkholderia cepacia complex and Burkholderia gladioli by automated ribotyping.

Several species belonging to the genus Burkholderia are clinically relevant, opportunistic pathogens that inhabit major environmental reservoirs. Consequently, the availability of means for adequate identification and epidemiological characterization of individual environmental or clinical isolates is mandatory. In the present communication we describe the use of the Riboprinter microbial characterization system (Qualicon, Warwick, United Kingdom) for automated ribotyping of 104 strains of Burkholderia species from diverse sources, including several publicly accessible collections. The main outcome of this analysis was that all strains were typeable and that strains of Burkholderia gladioli and of each species of the B. cepacia complex, including B. multivorans, B. stabilis, and B. vietnamiensis, were effectively discriminated. Furthermore, different ribotypes were discerned within each species. Ribotyping results were in general agreement with strain classification based on restriction fragment analysis of 16S ribosomal amplicons, but the resolution of ribotyping was much higher. This enabled automated molecular typing below the species level. Cluster analysis of the patterns obtained by ribotyping (riboprints) showed that within B. gladioli, B. multivorans, and B. cepacia genomovar VI, the different riboprints identified always clustered together. Riboprints of B. cepacia genomovars I and III, B. stabilis, and B. vietnamiensis did not show distinct clustering but rather exhibited the formation of loose assemblages within which several smaller, genomovar-specific clusters were delineated. Therefore, ribotyping proved useful for genomovar identification. Analysis of serial isolates from individual patients demonstrated that infection with a single ribotype had occurred, despite minor genetic differences that were detected by pulsed-field gel electrophoresis of DNA macrorestriction fragments. The automated approach allows very rapid and reliable identification and epidemiological characterization of strains and generates an easily manageable database suited for expansion with information on additional bacterial isolates.

Automation↗
PubMed2003

Validation of the automated reading and incubation system with Sensititre plates for antimicrobial susceptibility testing.

The present study compared the antimicrobial susceptibility testing (AST) results generated by the Automated Incubation and Reading System (ARIS) with custom Sensititre plates (TREK Diagnostic Systems, Cleveland, Ohio) and MicroScan PC10 GP and NUMIC10 GN plates interpreted with the WalkAway-96 system (Dade Behring, West Sacramento, Calif.) for gram-positive (GP) and gram-negative (GN) organisms as part of an in-house validation. A total of 326 isolates (3,689 antimicrobial agent-organism combinations) were evaluated. Sensititre plates were inoculated according to the instructions of the manufacturer with a suspension adjusted to a 0.5 McFarland standard, while the Prompt Inoculation System was used for the MicroScan plates. ARIS and the WalkAway system were used for automated reading of the Sensititre and MicroScan plates, respectively, at 18 to 24 h. The results were analyzed for essential (+/-1 twofold dilution) and categorical (sensitive, intermediate, or resistant) agreements. Plates that resulted in ARIS interpretations with major (falsely resistant) or very major (falsely susceptible) errors compared to the results obtained with the WalkAway system were read manually to corroborate instrument readings. Isolates for which very major or major errors were obtained and for which the results were not resolved by manual reading were retested in parallel. Isolates for which very major or major errors were obtained and for which the results were not resolved upon repeat testing were tested by the National Committee for Clinical Laboratory Standards M7-A5 frozen reference microdilution method. Essential agreement was 95.8% for 246 GN isolates. The following categorical error rates were obtained for the GN isolates: 1.3% minor errors, 0% major errors, and 0.4% very major errors. For 95 GP isolates, there was 93.5% essential agreement. Categorical error rates for GP isolates were 0.9% minor errors, 0.6% major errors, and 0.4% very major errors. ARIS-Sensititre is a diagnostic system feasible for use for automated AST in a clinical laboratory.

Automation↗
PubMed2003

Comparison of automated and manual nucleic acid extraction methods for detection of enterovirus RNA.

Automated nucleic acid extraction is an attractive alternative to labor-intensive manual methods. We compared two automated methods, the BioRobot M48 instrument (Qiagen, Inc.) and MagNA Pure (Roche Applied Sciences) methods, to two manual methods, the QIAamp Viral RNA Mini kit (Qiagen) and TRIzol (Invitrogen), for the extraction of enterovirus RNA. Analytical sensitivity was assessed by dilution analysis of poliovirus type 2 Sabin in cerebrospinal fluid. The sensitivity of PCR was equivalent after RNA extraction with QIAamp, BioRobot M48, and MagNA Pure. All 18 replicates of 100 PFU/ml were detected after extraction by the four methods. Fewer replicates of each successive dilution were detected after extraction by each method. At 10(-1) PFU/ml, 17 of 18 replicates were positive by QIAamp, 15 of 18 replicates were positive by BioRobot M48, and 12 of 18 replicates were positive by MagNA Pure; at 10(-2) PFU/ml, 4 of 17 replicates were positive by QIAamp, 2 of 18 replicates were positive by BioRobot M48, and 0 of 18 replicates were positive by MagNA Pure. At 10(-3) PFU/ml, no replicates were detected. Evaluation of TRIzol was discontinued after nine replicates due to a trend of lower sensitivity (at 10(-3) PFU/ml eight of nine replicates were positive at 100 PFU/ml, four of nine replicates were positive at 10(-1) PFU/ml, and zero of nine replicates were positive at 10(-2) PFU/ml). Concordant results were obtained in 24 of 28 clinical specimens after extraction by all methods. No evidence of contamination was observed after extraction by automated instruments. The data indicate that the sensitivity of enterovirus PCR is largely similar after extraction by QIAamp, BioRobot M48, and MagNA Pure; a trend of decreased sensitivity was observed after TRIzol extraction. However, the results of enterovirus PCR were largely concordant in patient samples, indicating that the four extraction methods are suitable for detection of enteroviruses in clinical specimens.

Automation↗
PubMed2004

Fully automated, internally controlled quantification of hepatitis B Virus DNA by real-time PCR by use of the MagNA Pure LC and LightCycler instruments.

We report on the development of a fully automated real-time PCR assay for the quantitative detection of hepatitis B virus (HBV) DNA in plasma with EDTA (EDTA plasma). The MagNA Pure LC instrument was used for automated DNA purification and automated preparation of PCR mixtures. Real-time PCR was performed on the LightCycler instrument. An internal amplification control was devised as a PCR competitor and was introduced into the assay at the stage of DNA purification to permit monitoring for sample adequacy. The detection limit of the assay was found to be 200 HBV DNA copies/ml, with a linear dynamic range of 8 orders of magnitude. When samples from the European Union Quality Control Concerted Action HBV Proficiency Panel 1999 were examined, the results were found to be in acceptable agreement with the HBV DNA concentrations of the panel members. In a clinical laboratory evaluation of 123 EDTA plasma samples, a significant correlation was found with the results obtained by the Roche HBV Monitor test on the Cobas Amplicor analyzer within the dynamic range of that system. In conclusion, the newly developed assay has a markedly reduced hands-on time, permits monitoring for sample adequacy, and is suitable for the quantitative detection of HBV DNA in plasma in a routine clinical laboratory.

Automation↗
PubMed2001

Introduction of an automated service for the laboratory confirmation of meningococcal disease in Scotland.

The Scottish Meningococcus and Pneumococcus Reference Laboratory provides a national service for the laboratory confirmation of meningococcal and pneumococcal disease in Scotland. The main tests used for the laboratory confirmation of meningococcal disease are culture, the polymerase chain reaction (PCR), antibody testing, and more recently DNA sequencing. This paper describes the automation of PCR for the laboratory confirmation of meningococcal disease and the typing of meningococcal isolates using DNA sequencing. Both methods have been automated using a robotic liquid handler and automated DNA sequencer. These methods, along with standard culture phenotyping and antibody testing, provide Scotland with an excellent service for the confirmation of meningococcal disease.

Automation↗
PubMed1994

Automated analysis of multiplex microsatellites.

The use of automated DNA fragment analysis with the Applied Biosystems 672 Genescanner system was evaluated in a routine diagnostic setting. The aim of the study was to compare automated fragment detection and analysis with conventional methods. For cystic fibrosis analysis the delta F508 mutation in exon 10 of the cystic fibrosis transmembrane regulator (CFTR) gene was multiplexed with two intragenic microsatellites. The analysis of the Prader-Willi/Angelman region of chromosome 15 used a panel of five microsatellites. For dystrophin, seven microsatellites covering the entire dystrophin gene were co-amplified. Automated analysis was faster and more accurate than analysis using radiolabelled products with sequencing gels, although some inconsistencies in the sizing of microsatellite alleles were seen.

Angelman Syndrome↗