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PubMed1992

One year's experiences with two different image analysis systems for automated reading of the contrast fluorescence test.

We have tested two different personal-computer based color image analysis systems for automated reading of the microlymphocytotoxicity test (LCT) for HLA-A,B,C-typing and screening. Over 17,000 single LCT-reactions were prepared using the simultaneous double fluorescent variant of the LCT (contrast fluorescence test, CFT). All tests were read visually by experienced laboratory staff members. For image analysis, an automated scanning system was used. In a first step, reactions were recorded on a videotape recorder using a color(CCD)-video camera. In a second step, the recorded reactions were analyzed with the two different image analysis systems by specifically developed programs. Good correlation (r = 0.89) of the score values assigned by digital image analysis with the visual tray reading was obtained. Since also the other main performance characteristics of the prototype system (throughput, reliability, compatibility) were acceptable for routine application, we may conclude that digital image analysis is a feasible and very interesting new technique for automated evaluation of the LCT.

Algorithms↗

PubMed2000

Evaluation of the BIOMIC video reader system for routine use in the clinical microbiology laboratory.

The BIOMIC Video Reader System (Giles Scientific, New York, NY, USA) is a semi-automated AST method that combines disk diffusion testing with automated reading and data interpretation. We evaluated this system with 497 strains comprising a total of 5821 drug organism combinations (DOC) from our routine diagnostic laboratory. Additionally, we compared the time required of the manual and the automated method. The overall agreement of interpretative categories of all DOC was 96.1%. However, comparing complete tests the agreement was only 70.8%. The average time required of the BIOMIC system to complete a test was more than twice as long as that of the manual method. Our data suggest that the tested version of the BIOMIC system cannot be recommended for routine use in diagnostic laboratories.

Automation↗

PubMedDate not supplied

Automated analysis with computerized information transfer in a university hospital.

The organization of a central clinical chemistry laboratory of a university hospital, which in 1992 analyzed 880,000 results, is presented. Automation with midsize selective analyzers, which run in parallel and are able to do positive sample identification on the primary bar-coded tubes, in connection with a laboratory computer system and a central hospital computer allowed for reliable and timely obtainment of results. The output of the results is made on laser printers in the medical wards, thus alleviating the pneumatic sample transport systems. Assessment of the turnaround time show that it can be kept below one hour. Such a setup without sorting out emergency demands allows a simplified sample flow. Thus, there is no need for a laboratory to make "stat" requests.

Autoanalysis↗

PubMed1997

A survey of the values of clinical chemistry parameters obtained for a common rat blood sample in ninety-eight Japanese laboratories.

A control survey was conducted to check the accuracy of automated analyzers used in the evaluation of clinical chemistry parameters in nonclinical toxicology studies. Pooled serum samples from male Sprague-Dawley rats were delivered refrigerated to each facility 98 laboratory facilities throughout Japan within 18 hours after sample preparation and analyzed. Commercially available normal human serum samples from a single lot were also analyzed at the same time. Survey results were divided into three categories. (1) Parameters with small coefficient of variation (CV) values for both rat and human serum samples included protein, glucose, cholesterol (CHO), urea nitrogen (UN), sodium (Na), potassium (K), chloride (Cl), calcium (Ca), and inorganic phosphate (IP). Definition of normal values in rats should be straight forward for these parameters. (2) Parameters with large CV values, but with a relatively good correlation between rat and human values include triglycerides (TG), glutamic oxaloacetic transaminase/aspartate aminotransferase (GOT/AST), glutamic pyruvic transaminase/alanine aminotransferase (GPT/ALT), and alkaline phosphatase (ALP). Measurements based on different principles gave different mean values, and this values contributed to the increase in CV values. Assessment of normal values would require a consideration of the measurement principles. (3) Parameters with large CV values only in rat serum samples included albumin (albumin/globulin ratio: A/G ratio), creatinine (CRE), and total bilirubin(BIL). Reactivity was different in rat albumin (ALB), depending on the reagents used. This difference needs to be corrected with values available by electrophoresis, or adjusted by rat ALB values, because of the lack of an appropriate measurement method. The enzyme method gave low values for rat CRE, which suggests the need for further examination of this method. The BIL values were extremely low in rat samples. It seems to be necessary to select appropriate methods to measure clinical pathology parameters correctly for rats. There was no deviation in values due solely to the mechanical operations of the analytical equipment. Non-standard initial settings of the equipment (equipment originally intended for human samples, but now applied to animal samples) was the main cause of the wide range of analytical values seen.

Albumins↗

PubMed1978

Prospective applications of calorimetry in the clinical laboratory.

Calorimetric analysis depends on the direct proportionality between the heat changes that occur during chemical reactions and the amount of reacting substances. Potential uses of calorimetry in the clinical laboratory are discussed, with examples. The calorimetric technique does not require optically clear specimens, and if the specificity of the measured reaction is assured, calorimetry can be used for quantitative determinations of components that are present in a complex matrix system such as body fluids. Specific enzymic reactions have been used to measure substrates and enzyme activities in biological specimens calorimetrically, with precision, sensitivity, and accuracy comparable to routine photometric techniques. The application of calorimetry in the clinical laboratory is limited now by its slowness, but development of automated instruments may enable the technique to become competitive with conventional analytical techniques in the clinical laboratory.

Calorimetry↗

PubMed1995

Quality assurance programmes in Portugal in the fields of clinical chemistry and haematology.

Quality assurance programmes in Portugal in the fields of clinical chemistry and haematology are running on under the organization of the correspondent laboratories of the National Institute of Health. They are carried out with voluntary participation for public and private clinical laboratories with confidentiality of results. The design comprehends two parts, each one supplementing the other: internal quality control, external quality assessment being the motivation for quality assurance usually done through courses and laboratory activities organized even before the laboratories enter the external scheme. In clinical chemistry: with six trials a year and 23 parameters to be analysed, the target values are assigned by reference laboratories. The survey reports involve a personal message and an Youden plot being the assessment of performance done either by a deviation index (DI) or, on enzymes, by a consensus value established among the participants. Yearly an individual message is produced to report the evolution of laboratory results. The state of the art shows a good performance of reference laboratories. The participant laboratories still evidence some difficulties on enzymes, bilirubin, creatinine, urates and urea determinations. In haematology the target value is the consensus median value of participant laboratories using the same kind of equipment, semi and totally automated. As survey reports two kinds of computer forms are produced: an individual message where each laboratory results are compared with the others through a DI, an yearly message with the DI evolution. Up to this date ten surveys a year are carried on for general haematology.(ABSTRACT TRUNCATED AT 250 WORDS)

Chemistry, Clinical↗

PubMed1999

[Precision and accuracy of white blood cell differentiation by an automated blood cell analyzer].

Precision and accuracy are important to assure the quality of clinical laboratory tests. We investigated the precision and accuracy of white blood cell (WBC) differentiation by automated blood cell analyzers. The coefficients of variation (CVs) of neutrophil, lymphocyte, monocyte, eosinophil and basophil differentiation by GEN.S (Coulter), based on a flow system and VCS technology, were 1.1%, 1.5%, 4.6%, 2.1%, and 33.3%, respectively. Between-run precision for neutrophil, lymphocyte, monocyte, eosinophil and basophil differentiation by GEN.S using cell control, 5C in 26 serial days was 2.0%, 2.8%, 6.5%, 9.2%, and 29.5%, respectively. The precision of WBC differentiation by blood cell analyzers based on the flow system was excellent except for basophil. We studied the accuracy of WBC differentiation by blood cell analyzers based on the flow system in contrast with eye-count. The correlations between neutrophil, lymphocyte, monocyte, eosinophil, and basophil percentages counted by GEN.S and by eye-count were r = 0.968, 0.971, 0.902, 0.940, and 0.391, respectively. The agreement rate between WBC differentiation by Microx, a blood cell analyzer based on pattern recognition, and eye-count cell by cell was good except for basophils. The accuracy of WBC differentiation by blood cell analyzers was also excellent excluding basophils.

Humans↗

PubMed2002

Discrepancy between visual and spectrophotometric assessment of sample haemolysis.

BACKGROUND: This study assessed the agreement between visual grading of sample haemolysis with automated haemolysis index (H index) measurement on the Roche 917 clinical chemistry analyser. METHODS: The H indices of 800 serum potassium samples and 800 EDTA plasma troponin I samples were compared with routine visual grading of sample haemolysis by technologists. Photographs of mild (haemoglobin 1 g/L), moderate (2.5 g/L) and severe (5 g/L) haemolysis were available for reference. RESULTS: For serum samples, the weighted kappa coefficient was 0.42 with an 8.0% haemolysis rate determined by visual inspection and 3.4% by H index. For EDTA-plasma samples, the weighted kappa coefficient was 0.35 with a 4.5% haemolysis rate determined by visual inspection and 7.9% by H index. CONCLUSION: Visual assessment of sample haemolysis is unreliable and can give variable results depending on sample type. Laboratories should consider investigating sample haemolysis using automated haemolysis index measurement as an alternative to visual grading whenever it is available.

Clinical Laboratory Techniques↗

PubMed2003

Serum insulin-like growth factor I reference values for an automated chemiluminescence immunoassay system: results from a multicenter study.

BACKGROUND: Analysis of insulin-like growth factor I in serum (S-IGF-I) is an integral component in the diagnosis of GH-related disorders and is going to be of interest in the diagnosis and follow-up of many disorders. The objective of the present study was to develop cross-sectional reference values for S-IGF-I measured by an automated chemiluminescence immunoassay (Nichols Advantage). METHODS: The study included samples from 3,961 healthy subjects (2,201 males, 1,760 females) aged 1 month to 88 years. Six laboratories were involved in this study and the samples were analyzed by one of seven automated immunoassay systems run in these laboratories. For data analysis, polynomial age and sex-specific models were fitted after transformation of S-IGF-I values. RESULTS: The results show the well-known age dependency of S-IGF-I levels. At ages <20, higher S-IGF-I levels were seen in girls with an estimated mean peak of 410 microg/l at age 14 and an estimated mean peak of 382 microg/l at age 16 in boys. Thereafter, a rapid decrease was seen to approximately 25 years of age, followed by a slow age-dependent decrease. In adulthood, S-IGF-I in males were slightly, but significantly higher than in females. It could be shown that the mean values of some reference sample subgroups differed significantly from the total mean. However, the multicenter approach used in this study reduces the impact of systematic population, sample handling and laboratory differences on the calculated reference mean. CONCLUSION: The present study establishes age- and sex-specific reference values for a fully automated immunoassay system based on a large population of healthy subjects. The established reference values may be used for this immunoassay system in different laboratories provided that the systematic difference between systems is low.

Adolescent↗

PubMed1999

A fully-automated environmental chamber for examination of long-term effects of intermittent hypoxia on medium-size animals.

We describe the design and construction of a fully-automated environmental chamber for the simultaneous exposure of up to four medium-size laboratory animals to long-term intermittent hypoxia. The air-sealed automated environmental chamber consists of a box equipped with a ventilation fan and three electrically-activated solenoid valves. Our system was used to expose four rabbits to 12 h of repetitive episodes of hypoxia (environmental O2 concentration 12-13%) lasting 45 min followed by breathing room air for 15 min. During environmental hypoxia, the mean arterial PaO2 and PaCO2 were 41 +/- 3.0 and 24 +/- 0.7 mmHg (mean +/- SEM), respectively. In this system, opening and closing of the solenoid valves is fully computerized to allow different settings of the duration and severity of hypoxia. The chamber is safe and fully automated and cost-effective for studying the effects of long-term intermittent hypoxemia in medium-size animals.

Animals↗

PubMed1995

The illuminated high contrast macular grid: a pilot study.

BACKGROUND: Analysis of visual disturbances in the central 10 degrees provides valuable data for the low vision clinician. Amsler grid testing in the visually impaired can be poorly visible and it can be difficult to transcribe results. Automated perimeters will accurately map retinal defects, but the equipment is expensive and this technique is time consuming. An Amsler grid was modified using principles of luminance and contrast to improve its accuracy, and its efficacy was compared to an automated perimeter. METHODS: Eighteen visually impaired patients (28 eyes) were prospectively evaluated at the Emory Low Vision Laboratory. Each patient had three central threshold visual field tests: automated perimetry (Humphrey model 690), conventional Amsler Grid, and Illuminated High Contrast Macular Grid (IHCMG). The Humphrey central 10-2 threshold automated visual field examination was performed to map the patient's retinal function. RESULTS: The Humphrey analysis identified (mean +/- one standard deviation) 2.4 +/- 1.4 defects per patient with an average percent defect area of 33.4 +/- 14.8 degrees. The IHCMG technique was not significantly different from the Humphrey technique for identifying the number of defects (2.25 +/- 1.5) (p > 0.05) or the percent defect area (26 degrees +/- 20 degrees) (p > 0.05). The conventional Amsler grid technique significantly under-evaluated the retinal defects compared to the Humphrey analysis of the number of defects (p < 0.01) and percent area of defects (p > 0.001). CONCLUSIONS: The IHCMG technique can be used as a simple, rapid and accurate analysis of the central 10 degrees visual field in the low vision patient.

Aged↗

PubMed1983

OpScan-MIMS. A data network system.

The expansion of a unique system for medical, epidemiological, and laboratory information management is described. The system utilizes an automated, optically scanned data entry mode (OpScan), and a generalized, interactive storage and retrieval software package (MIMS). Aggregate medical, epidemiological, and laboratory information are monitored, with the permission of the locality, via telephone terminals at four sexually transmitted disease clinics. OpScan-MIMS combines an efficient, rapid, and cost-effective method for data entry with a user-friendly computer system that requires no specialized training in computer language.

Centers for Disease Control and Prevention, U.S.↗

PubMed1979

An evaluation of two methods of laboratory quality control.

Two methods for quality control of automated blood counters have been compared: (1) monitoring results from patients' samples, and (2) analysis of results of a stable whole-blood control. The latter method proved better able to distinguish calibration changes from patient variation.

Erythrocyte Indices↗

PubMed2001

Pharmacogenetics: the Dx perspective.

The promise of the rapidly developing field of pharmacogenetics is that genetically determined propensities of individual patients to respond favorably or adversely to a given pharmacologic agent will be able to be determined prior to administration of that drug. The realization of that promise, however, is predicated on a number of developments in the capabilities of diagnostic laboratories. These developments include the introduction of automated technologies for efficiently and accurately detecting, quantifying and decoding specific nucleic acid sequences and the concomitant availability of information technology-based applications for rapidly analyzing, interpreting and then communicating complex genetic data to healthcare providers. This article will review currently available and developmental molecular diagnostic technology and in addition, describe the current status and speculate on the future of pharmacogenetic testing in the clinical laboratory.

Humans↗

PubMed2001

Use of automated microscopy for the detection of disseminated tumor cells in bone marrow samples.

The use of automated microscopy has reached the maturity necessary for its routine use in the clinical pathology laboratory. In the following study we compared the performance of an automated microscope system (MDS) with manual method for the detection and analysis of disseminated tumor cells present in bone marrow preparations from breast carcinoma patients. The MDS System detected rare disseminated tumor cells among bone marrow mononuclear cells with higher sensitivity than standard manual microscopy. Automated microscopy also proved to be a method of high reproducibility and precision, the advantage of which was clearly illustrated by problems of variability in manual screening. Accumulated results from two pathologists who had screened 120 clinical slides from breast cancer patients both by manual microscopy and by use of the MDS System revealed only two (3.8%) missed by the automatic procedure, whereas as many as 20 out of 52 positive samples (38%) were missed by manual screening.

Bone Marrow Examination↗

PubMed2004

A vertical flow chamber for Xenopus oocyte electrophysiology and automated drug screening.

Xenopus laevis oocytes are used extensively in the study of ion channel coupled receptors. Efficient use of oocytes for ion channel characterization requires a system that is inherently stable, reproducible, minimizes drug volumes, and maximizes oocyte longevity. We have constructed a vertical flow oocyte recording chamber to address the aforesaid issues, where the oocyte is placed in a funnel-shaped chamber and perfused from the bottom of the funnel. The vertical rather than horizontal flow of perfusate results in an unusually stable environment for oocyte recording. Two-electrode voltage clamp recordings from a single oocyte are acquired easily and routinely over several hours while maintaining stable baseline currents and reproducible response profiles. Chamber characteristics were tested using a serotonin ligand-gated ion channel receptor (5-HT3R). Data obtained from this system corresponds well with published data. To further test the stability and reliability of this perfusion chamber, we constructed an automated oocyte perfusion system utilizing a commonly available HPLC autosampler. We were able to obtain dose-response curves for various 5-HT3AR ligands using the automated perfusion system with minimal user intervention. Such a system can easily satisfy need for automated oocyte electrophysiology in academic settings, especially small to medium sized laboratories.

Animals↗

PubMed1994

Haemophilia A diagnosis by automated fluorescent DNA detection of ten factor VIII intron 13 dinucleotide repeat alleles.

Haemophilia A is a recessive X linked bleeding disorder caused by deficiency or functional abnormality of coagulation factor VIII. This disease usually has no visible phenotype in female carriers; hence, great efforts are made to offer all haemophilia A families accurate carrier diagnosis. Significant progress in this direction was made with the identification of the intron 13 variable number tandem repeat (VNTR), which is hitherto the most informative single marker within the factor VIII gene. The authors have established intron 13 VNTR detection in their laboratory by adapting its analysis to an automated sequencer using different primers of which one is fluorescent dye labelled. With this method, which is more rapid and convenient than that originally described, 67 haemophilia A families of German origin were screened and two new alleles (alleles 17 and 25) were identified. The informativeness of the VNTR in these families based on the patients maternal X chromosomes (134) is about 67%.

Alleles↗

PubMed2001

Detection of mutations in the dystrophin gene via automated DHPLC screening and direct sequencing.

BACKGROUND: Currently molecular diagnostic laboratories focus only on the identification of large deletion and duplication mutations (spanning one exon or more) for Duchenne Muscular Dystrophy (DMD) yielding 65% of causative mutations. These mutations are detected by an existing set of multiplexed polymerase chain reaction (PCR) primer pairs. Due to the large size of the dystrophin gene (79 exons), finding point mutations (substitutions, deletions or insertions of one or several nucleotides) has been prohibitively expensive and laborious. The aim of this project was to develop an effective and convenient method of finding all, or most, mutations in the dystrophin gene with only a moderate increase in cost. RESULTS: Using denaturing high performance liquid chromatography (DHPLC) screening and direct sequencing, 86 PCR amplicons of genomic DNA from the dystrophin gene were screened for mutations in eight patients diagnosed with DMD who had tested negative for large DNA rearragements. Mutations likely to be disease-causative were found in six of the eight patients. All 86 amplicons from the two patients in whom no likely disease-causative mutations were found were completely sequenced and only polymorphisms were found. CONCLUSIONS: We have shown that it is now feasible for clinical laboratories to begin testing for both point mutations and large deletions/duplications in the dystrophin gene. The detection rate will rise from 65% to greater than 92% with only a moderate increase in cost.

Automation↗