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PubMedDate not supplied

Comparison of manual and automated methods of liquid-based cytology. A morphologic study.

OBJECTIVE: To evaluate the morphologic characteristics of gynecologic samples prepared by 3 different methods of liquid-based cytology. STUDY DESIGN: Cytologic samples from representative cases of each diagnostic category of squamous epithelial lesion, prepared by automated and manual liquid-based systems, were analyzed by 3 laboratories in the United States, Portugal and Brazil. The systems included: ThinPrep (automated, U.S. Food and Drug Administration approved; Cytyc Corp., Boxborough, Massachusetts, U.S.A.), Autocyte PREP (South American system, manual; TriPath Imaging, Inc., Burlington, North Carolina, U.S.A.) and DNACITOLIQ (manual; Digene Brazil, São Paulo, Brazil). A panel of 16 morphologic parameters was evaluated: cellularity, clean background, uniform distribution, artifacts, cellular overlapping, architectural and cytoplasmic distortion, cytoplasmic vacuolization, cellular elongation, imprecise cytoplasmic borders, folded cytoplasmic borders, nuclear hyperchromasia, coarse chromatin, prominent nucleolus, irregular nuclear borders, atypical mitosis and inflammatory infiltrate. Negative, atypical squamous cells of undetermined significance, low grade squamous intraepithelial lesion (LSIL) and high grade squamous intraepithelial lesion (HSIL) cases were included. Cases without biopsies were confirmed by consensus. RESULTS: Cellularity was adequate in all samples. Clean background was observed in the vast majority of samples with all liquid-based systems. Uniform distribution was frequently found in ThinPrep and Autocyte PREP samples but not in DNACITOLIQ. Artifacts were not present in DNACITOLIQ samples, rare in ThinPrep and observed in 8 (34.7%) Autocyte PREP. Cellular overlapping was observed in all 3 system samples: 11 (31.42%) cases in ThinPrep, 16 (69.56%) in Autocyte PREP and 17 (68%) in DNACITOLIQ System. Architectural and cytoplasmic distortion were present in 3 cases of HSIL (13%) and cytoplasmic vacuolization in 2 cases of LSIL and 1 HSIL of Autocyte PREP. Cellular elongation was found in 13 (56.5%) Autocyte PREP and in 5 (20%) DNACITOLIQ samples. Inflammatory infiltrate was found in all 3 system samples but with more frequency in the Autocyte PREP (69.56%) and DNACITOLIQ System (72%). CONCLUSION: This study clearly indicated that in spite of the different methodologies, the 3 methods adequately preserved cellular structure for morphologic evaluation. The choice of method will depend on price, availability and procedures involved.

Artifacts↗
PubMed2002

Rokitamycin: bacterial resistance to a 16-membered ring macrolide differs from that to 14- and 15-membered ring macrolides.

Rokitamycin is the latest semi-synthetic 16-membered ring macrolide introduced into clinical practice. It is characterized by greater hydrophobicity, better bacterial uptake and a slower release, more cohesive ribosomal binding, and a longer post-antibiotic-effect (PAE) than can be observed with other available 14-, 15- and 16-membered ring macrolides. Rokitamycin exerts its activity on strains that harbor inducible erm genes or the efflux gene, mef(A). It has also been reported to be more active in vitro than other 16-membered ring macrolides. However, these recognized features are not fully exploited yet because current automated test procedures used in many microbiological laboratories determine susceptibility only to erythromycin or clarithromycin. Resistance to 16-membered ring macrolides cannot be predicted solely on the basis of known resistance to erythromycin or clarithromycin as revealed by an automated susceptibility assay. At least equally important is the knowledge of the bacterial resistance phenotype. This is underlined by the existence of Gram-positive coccal strains resistant to erythromycin and other 14-,15-membered ring macrolides but susceptible to 16-membered ring macrolides. Since the local prevalence of erythromycin phenotypes is generally unknown but might determine the outcome of treatment, the procedure for identifying the phenotypes in erythromycin-resistant strains (which can be easily and cheaply performed using the two- or three-disk assay) should become routine, at least in the countries in which 16-membered ring macrolides are used. This approach should help to optimize the use of macrolides, improve our knowledge of the local prevalence of phenotypes resistant to erythromycin, and offer the possibility of treating infections caused by certain types of erythromycin-resistant pathogens.

Anti-Bacterial Agents↗
PubMed1991

Automated procedure for dewaxing and rehydration of paraffin-embedded tissue sections for DNA flow cytometric analysis of breast tumors.

Flow cytometric DNA analysis of paraffin-embedded tumors is an important diagnostic and prognostic tool in clinical pathology. The technique is limited, however, by the time-consuming multistep procedure for dewaxing and rehydrating tissue. The authors developed an automated procedure to complete the dewaxing and rehydration of tissue using a routine histologic tissue processor with a 24-hour timer. This technique provided excellent tissue recovery and reproducible DNA histograms comparable to those obtained by manual methods. Subsequently, the authors analyzed the DNA content of 93 paraffin-embedded breast cancer tissues. The automation of a significant portion of the routine processing required for paraffin-embedded tissue makes cytometric DNA analysis a more practical procedure in the laboratory.

Automation↗
PubMedDate not supplied

The learning department, Part two.

Whether you want to move deliberately toward becoming a learning department or not, the kind of organization that is described here is probably appealing and a little frightening. The staff is more self-reliant, there is a higher level of motivation, communication is more open, there is a dedication to continuous improvement, and there is more intentional, "smart" risk-taking. At the beginning of our last column ("The Learning Department, Part One"), we listed the five trends that we believe will shape the clinical laboratory of the future: 1. increased automation and the attendant reduction and shift in the technical skills required 2. chronic shortages of qualified staff 3. changes in the legislation that affects credentialing and regulation 4. expanded uses of molecular diagnostics 5. emergence of the lab as a major contributor to the hospital's bottom line. It is hard to imagine surviving these turbulent trends without instilling many of the characteristics of a learning department in your organization. What are the most important steps to take? Open communications, schedule debriefs to learn from your experiences, and stay curious!

Career Mobility↗
PubMed2002

Diagnosis of fungal infection: new technologies for the mycology laboratory.

The dramatic increase in nosocomial invasive mycoses over the past two decades has led to increased interest in the area of antifungal development. Unfortunately, the infusion of new diagnostic technology into the clinical mycology laboratory has lagged behind. Although newer, automated, continuous-monitoring blood culture systems are as sensitive as the older, manual "gold standard" system, the recovery of fungi from blood, as well as other clinical specimens, remains an insensitive marker for invasive fungal infection. Antigen assays for the rapid diagnosis of invasive fungal infections are in development, and galactomannan and glucan are two such promising antigens. Glucan may be present in the blood of patients with infection secondary to a wide variety of fungal pathogens, including Candida, Aspergillus, Fusarium, Saccharomyces, Trichosporon and Acremonium species. Early data suggest galactomannan may be present in the blood in detectable levels very early in the course of invasive aspergillosis. The galactomannan assay currently undergoing evaluations may actually be positive prior to the clinical suspicion for infection and may be useful in monitoring therapeutic response as well; however, the etiology of false-positive results following cytotoxic chemotherapy still has to be elucidated. PCR assays are also being developed in the research laboratory, however, the PCR assays will require a significant amount of adaptation and validation before they are ready for clinical care. Well-planned studies to evaluate the performance characteristics as well as appropriate clinical and cost-effective application of these new tests are needed.

Antigens, Fungal↗
PubMed2006

Repeatability of nerve conduction measurements using automation.

OBJECTIVE: To quantify nerve conduction study (NCS) reproducibility utilizing an automated NCS system (NC-stat, NeuroMetrix, Inc.). METHOD: Healthy volunteers without neuropathic symptoms participated in the study. Their median, ulnar, peroneal, and tibial nerves were tested twice (7 days apart) by the same technician with an NC-stat instrument. Pre-fabricated electrode arrays specific to each nerve were used. Both motor responses (compound motor action potential [CMAP] and F-waves - all nerves) and sensory responses (sensory nerve action potentials [SNAP] - median and ulnar nerves only) were recorded following supramaximal stimuli. Automated algorithms determined all NCS parameters: distal motor latency (DML), mean F-wave latency (FWL), distal sensory latency (DSL), CMAP amplitude, and SNAP amplitude. Latency was adjusted for skin temperature deviation from reference. Pearson correlation coefficient (CC), intraclass correlation coefficient (ICC), coefficient of variance (CoV), and relative intertrial variation (RIV) were calculated. RESULTS: Fifteen subjects participated in either upper or lower extremity studies with nine participating in both. With the exception of CMAP amplitude, all parameters had CoV less than 0.06. Upper extremity amplitude parameters had CCs greater than 0.85. CCs for latencies were greater than 0.80 except for the median nerve FWL (CC = 0.69). For lower extremity nerves, ICCs were highest for mean FWL (>0.90), followed by DML (>0.82) and then CMAP (peroneal 0.33, tibial 0.73). The 10th to 90th RIV percentiles were bounded by +/-7% for F-wave latencies; +/- 9% for all DSLs; and +/- 11% for DML (except peroneal at 15%). CONCLUSIONS: The reproducibility of NCS parameters obtained with an automated NCS instrument compared favorably with traditional electromyography laboratories. F-wave latencies had the highest repeatability, followed by DML, DSL, SNAP and CMAP amplitude. Given their high reproducibility, automated NCS instrument may encourage wider utilization of NCS in clinical and research applications.

Action Potentials↗
PubMed2004

Preliminary assessment of an automated surveillance system for infection control.

BACKGROUND AND OBJECTIVE: Rapid identification and investigation of potential outbreaks is key to limiting transmission in the healthcare setting. Manual review of laboratory results remains a cumbersome, time-consuming task for infection control practitioners (ICPs). Computer-automated techniques have shown promise for improving the efficiency and accuracy of surveillance. We examined the use of automated control charts, provided by an automated surveillance system, for detection of potential outbreaks. SETTING: A 656-bed academic medical center. METHODS: We retrospectively reviewed 13 months (November 2001 through November 2002) of laboratory-patient data, comparing an automated surveillance application with standard infection control practices. We evaluated positive predictive value, sensitivity, and time required to investigate the alerts. An ICP created 75 control charts. A standardized case investigation form was developed to evaluate each alert for the likelihood of nosocomial transmission based on temporal and spatial overlap and culture results. RESULTS: The 75 control charts were created in 75 minutes and 18 alerts fired above the 3-sigma level. These were independently reviewed by an ICP and associate hospital epidemiologist. The review process required an average of 20 minutes per alert and the kappa score between the reviewers was 0.82. Eleven of the 18 alerts were determined to be potential outbreaks, yielding a positive predictive value of 0.61. Routine surveillance identified 5 of these 11 alerts during this time period. CONCLUSION: Automated surveillance with user-definable control charts for cluster identification was more sensitive than routine methods and is capable of operating with high specificity and positive predictive value in a time-efficient manner.

Cross Infection↗
PubMed2005

Interlaboratory validation of a CD71-based flow cytometric method (Microflow) for the scoring of micronucleated reticulocytes in mouse peripheral blood.

An interlaboratory study was performed to validate an anti-CD71/flow cytometry-based technique for enumerating micronucleated reticulocytes (MN-RETs) in mouse peripheral blood. These experiments were designed to address International Workshop on Genotoxicity Test Procedures validation criteria by evaluating the degree of correspondence between MN-RET measurements generated by flow cytometry (FCM) with those obtained using traditional microscopy-based methods. In addition to these cross-methods data, flow cytometric MN-RET measurements for each blood sample were performed at two separate sites in order to evaluate the reproducibility of data between laboratories. In these studies, groups of male CD-1 mice were treated with vehicle (saline or vegetable oil), a negative control (saline or vegetable oil), or four dose levels of five known genotoxicants (clastogens: cyclophosphamide, benzo[a]pyrene, 5-fluorouracil, methotrexate; aneugen: vincristine sulfate). Exposure occurred on 3 consecutive days via intraperitoneal injection, and blood samples were obtained approximately 24 hr after the final treatment. MN-RET frequencies were determined for each sample based on the analysis of 2,000 (microscopy) and 20,000 (FCM) reticulocytes. Regardless of the method utilized, each genotoxic agent was observed to cause statistically significant increases in the frequency of MN-RETs, and each response occurred in a dose-dependent manner. Spearman's correlation coefficient (rs) for FCM versus microscopy-based MN-RET measurements (nine experiments, 252 paired measurements) was 0.740, indicating a high degree of correspondence between methods. The rs value for all flow cytometric MN-RET measurements performed at the two independent sites was 0.857 (n = 248), suggesting that the automated method is highly transferable between laboratories. Additionally, the flow cytometric system offered advantages relative to microscopy-based scoring, including a greater number of cells analyzed, much faster analysis times, and a greater degree of objectivity. Collectively, data presented in this report suggest that the overall performance of mouse peripheral blood micronucleus tests is enhanced by the use of the flow cytometric scoring procedure.

Animals↗
PubMed1999

Quality control in a manual and an automated leukocyte differential count.

Quality control (QC) has been introduced in laboratories, and QC surveys in leukocyte differential count to enhance quality have been performed by College of American Pathologists, Japanese Association of Medical Technologists, Osaka Medical Association and manufacturers. The results of QC survey in a manual leukocyte differential count indicated problems on the differentiation of segmented neutrophils and band neutrophils and the detection of pathological blood cells on blood smear. While the results of QC survey in an automated leukocyte differential count performed by same manufacturer with an automated blood cell counter were satisfactory, however, there was a difference in leukocyte differential cell counts among laboratories with other manufacturer's instruments because the synthetic blood material used in QC is an exclusive item for an instrument. It is necessary to further reeducate the medical technologists in order to improve morphological performance, and to standardize the synthetic blood material for compatibility with various automated blood cell counters.

Automation↗
PubMed1975

Laboratory experience with a radiometric method for detecting bacteremia.

Two bacteriologic systems for detecting bacteria in blood were compared; the automated radiometric BACTEC and the conventional method used in our laboratory for many years. BACTEC consisted of two bottles with 30 ml and the conventional method with 50 ml of media for aerobes and anaerobes. The BACTEC bottles were inoculated with 2 to 3 ml and the conventional with 4 to 5 ml of blood at the patient's bedside. Out of the 3,045 blood specimens cultured (804 patients), 262 (117 patients) were positive by one or both methods. The conventional system detected 5more cultures. The explanation of the differences is discussed. Positive blood cultures were detected by the BACTEC procedure as early as 6 h after the blood collection. In the first 24 h, on the average, 77% of aerobic organisms were detected by the BACTEC as compared to 48% by the conventional system. All anaerobic BACTEC cultures were positive within 4 days, whereas the conventional system detected at that time 74%. At day 4, 67% of fungi were detected by the BACTEC and only 27% by the conventional system. Of the 3,045 blood cultures examined by the BACTEC, 208 were recorded as false positive with growth index readings ranging from 30 to 59.

Aerobiosis↗
PubMed1990

Methodical and ethical aspects of nucleic acid diagnostics. A review.

Aside from the established enzymatic and immunodiagnostic procedures, an increasing number of diagnostic procedures for nucleic acid detection--i.e. for the detection of the primary genetic information--has been developed in recent years. For the routine use of these diagnostic procedures in central analytical laboratories or by the general practitioner, automated DNA analysis methods with integrated highly sensitive detection systems have been developed. The most important objective of these developments--aside from the elucidation of the type of genetic defect on the molecular level through basic research--is the generation of a quantitative nonradioactive signal in corresponding analyzers. Detection methods using the ELISA principle as quantifiable reporter system--analogous to the automatic immunodiagnostic procedures--have hence been developed in recent years. Owing to the increasing importance of this analysis procedure alternative to the detection of bioactive low molecular weight substances or proteins--i.e. nucleic acid diagnostics--the following is an attempt to provide a survey of the methods and possibilities of application. Moreover, possibilities for nonradioactive signal generation and amplification will be presented. The chances and risks of genome diagnostics will be discussed in a final section.

Animals↗
PubMedDate not supplied

Fundamentals of computerised data acquisition in the human performance laboratory.

Researchers of the future will depend upon computer technology to address questions that previously could not be answered using traditional methods of data acquisition. Recent advances in computer design and software availability have eliminated many of the limitations previously associated with computer use in the laboratory setting. Although computerisation facilitates the acquisition of laboratory data, identification of errors introduced into the measurement is often more difficult. The laboratory computer is only one component of an automated data acquisition system. A system designed to collect physiological data comprises several primary components, including: (a) sensing element; (b) signal amplification/conditioning circuits; (c) analogue display; (d) computer interface; (e) laboratory computer; and (f) acquisition software. The acquisition and conditioning of analogue signals is accomplished using traditional laboratory procedures familiar to the researcher. Converting this analogue signal into a digital format used by the computer, however, represents new techniques. Fundamentals of computerised data acquisition are discussed. An understanding of how an automated data acquisition system is configured and the techniques used to transform the values will assist the user in identifying sources of error introduced into the measurement. Additionally, familiarization with the methods of computerisation will provide insight into future applications of computer technology in the human performance laboratory.

Analog-Digital Conversion↗
PubMed2004

Noncompetitive enzyme immunoassay for carcinoembryonic antigen by flow injection chemiluminescence.

BACKGROUND: Recently, many automated immunoassay analyzers have been developed for carcinoembryonic antigen (CEA) to overcome the shortcomings in traditional immunoassay methods that are time-consuming and labor-intensive. Flow injection immunoassay (FIIA) has been increasingly applied to laboratory medicine due to its ease in automation, rapid speed and reproducible results. It is important to develop a FIIA method for CEA determination. METHODS: Based on a noncompetitive immunoassay format, a CEA-immobilized immunoaffinity column inserted in the flow system was used to trap the unbound horseradish peroxidase (HRP)-labeled antibody after an off-line incubation of CEA and HRP-labeled anti-CEA. The trapped enzyme conjugate was detected by injecting substrates to produce an enhanced chemiluminescence (CL). RESULTS: The linear range for CEA was 1.0-25 ng/ml with a correlation coefficient of 0.997 and a detection limit of 0.5 ng/ml. The sampling and chemiluminescence detection time for one sample was 5 min after a preincubation procedure of 25 min. Twenty five human serum samples detected by this method were in good agreement with the results obtained by immunoradiometric assay (IRMA). CONCLUSIONS: This method could be used for rapid analysis of CEA and potentially other antigens.

Carcinoembryonic Antigen↗
PubMed2026

AI'm Here to Help: Enhancing Laboratory Genetic Counseling with Artificial Intelligence.

BACKGROUND: Artificial intelligence (AI) is transforming the fields of genetics and genetic counseling, enhancing both clinical and laboratory practices. The rise of AI technologies has drawn attention to their potential impact on genetic counseling, particularly in patient diagnosis and the counseling processes. CONTENT: In the laboratory, AI plays a critical role in improving communication between laboratory genetic counselors and healthcare providers by automating routine tasks and optimizing workflows. These advancements allow genetic counselors to dedicate more time to addressing complex inquiries, improving genetic test selection, and helping providers interpret genetic test results. As AI continues to integrate into laboratory genetic counseling practice, it presents both opportunities and challenges. At the time of submission, there is a large knowledge gap regarding AI and its application to laboratory genetic counseling, given the lack of published information on this topic. SUMMARY: This article summarizes existing literature, the history and current applications of AI in laboratory genetic counseling, examines its benefits and limitations, and explores future directions for its implementation in the field.

Humans↗
PubMed1999

Quality control in urinalysis.

Quality control (QC) has been introduced in laboratories, and QC surveys in urinalysis have been performed by College of American Pathologist, by Japanese Association of Medical Technologists, by Osaka Medical Association and by manufacturers. QC survey in urinalysis for synthetic urine by the reagent strip and instrument made in same manufacturer, and by an automated urine cell analyser provided satisfactory results among laboratories. QC survey in urinalysis for synthetic urine by the reagent strips and instruments made by various manufacturers indicated differences in the determination values among manufacturers, and between manual and automated methods because the reagent strips and instruments have different characteristics, respectively. QC photo survey in urinalysis on the microscopic photos of urine sediment constituents indicated differences in the identification of cells among laboratories. From the results, it is necessary to standardize a reagent strip method, manual and automated methods, and synthetic urine.

Automation↗
PubMed1993

[The pitfalls of lymphocyte immunophenotyping during HIV infection].

Preferential infection of CD4+ lymphocytes by HIV, related to both direct and indirect mechanisms, leads to progressive decrease in this population. A CD4 lymphocyte count has become the preferential test for monitoring the infection. Thus, a flow cytometry laboratory handles a great number of phenotyping prescriptions that require accuracy and precision. The technical and biological parameters involved are listed, and some rules for interpretation of results given. Because no standardization exists and because cytometers are not automated, there is often great variability between laboratory results. We show that variability can be decreased by the use of correct procedures for analysis on the cytometer (especially for fluorescence interpretation). On the other hand, many biological factors have a significant effect on the results. A list of these factors is given: circadian rhythm, other infectious diseases and drugs. Lastly, the distribution of values in normal subjects is reported. The wide range shows that results from a patient must be compared with previous values to determine the course of the infection. These data would help in defining and interpreting results.

Adult↗
PubMed1983

[Chances, possibilities and effects of new installations, mainly automated protocols in the field of cytology as well as cytologic early cancer diagnosis].

In the recent past symptoms of a reorganization of laboratory medicine became apparent. They were brought about by methodological progress in analytical biochemistry, chemistry and also by the rapidly increasing efficiency of automation in the clinical laboratory as well as in medical practice. A somewhat parallel change also comes into existence in cytology and also, within certain limits, the histology. Since the early seventies the German Government, in accord with these developments, has supported some projects purporting to design better, more reproducible and more simple procedures in clinical diagnosis. With regard to cytological detection of malignancy this meant the development of methods to render cytological diagnosis a more potent tool for the recognition of cancer at an earlier stage on pre-selected material and with a high degree of sensitivity. Requirements on the quality of these methods with regard to reliability and safety will be discussed together with those marginal conditions which are expected to have a bearing on decisions aiming at the introduction of those methods into cytological routine. Consideration of those problems will be based on qualification requirements on prescreening procedures in general and manual cytological prescreening in particular. Discussion will also be extended on procedural sources of error.

Autoanalysis↗