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Determination of catecholamines in plasma by HPLC and amperometric detection. Comparison with a radioenzymatic method.

The determination of norepinephrine and epinephrine in plasma by HPLC with amperometric detection was modified, giving detection limits of 25 ng/l for norepinephrine and epinephrine, respectively, using 1 ml plasma. In order to achieve this sensitivity, it was necessary to minimize the background noise by modification of instrumentation and specimen handling. Particularly important was the extra purification of the reagents, the application of micro-bore HPLC, the enzymatic cleavage of uric acid and temperature control of the amperometric cell and the amplifier. Comparison of the present method with the radioenzymatic determination of catecholamines resulted in coefficients of correlation of r = 0.924 and 0.919 for norepinephrine and epinephrine, resp. (n = 38). The concentrations of the 38 different samples used for the comparison were in the physiological range.

Catecholamines↗

The suspensory ligament of the endolymphatic duct in the rat. An ultrastructural study.

Following optimized fixation and specimen handling, the endolymphatic duct was investigated by transmission and scanning electron microscopy. Tubular microfibrils, 10-12 nm thick and of indefinite length, occupied the subepithelial compartment abundantly and seemed to insert into the basal lamina of the epithelial cells of the duct and to merge with the bony wall of vestibular aqueduct thus forming a suspensory ligament of the endolymphatic duct. This ligament is suggested to protect the endolymphtaic duct from collapsing and to play a role in the mechanical protection of the vestibular apparatus from sudden increases in intracranial pressure. The capillaries and the aqueductal vein were located in intimate relationship to the epithelial cells. Both types of vessels were provided with fenestrations of the endothelial cells, supporting the proposal that the endolymphatic duct may be involved in the resorption of endolymph.

Animals↗

Problems with the laboratory identification and evaluation of coagulase-negative staphylococci.

During a 105 day period in mid 1984, 796 isolates of coagulase-negative staphylococci were recovered from routine specimens handled in the Microbiology Laboratory, Dunedin Hospital. Of these isolates 504 (63.3%) were from wounds, 170 (21.4%) from urines, 58 (7.3%) from intravascular catheter tips, 44 (5.5%) from blood cultures and 20 (2.5%) from sputa. Presumptive identification of 315 consecutive isolates revealed 175 (55.6% of total) strains of Staphylococcus epidermidis, 44 (14.0%) S. capitis, 36 (11.4%) S. haemolyticus/hominis, 29 (9.2%) S. warneri, 19 (6.0%) S. simulans and 12 (3.8%) members of the S. saprophyticus group. Using laboratory generated criteria, 44.8%, 25.9% and 4.5% of coagulase-negative isolates from catheter tips, urines and blood cultures respectively, were deemed pathologically potentially significant. Although more common, S. epidermidis did not appear to be significantly more virulent than other members of the epidermidis species group or S. simulans; 67% of the S. saprophyticus group isolates from urine were considered pathologically significant. Antibiograms on the 796 coagulase-negative isolates revealed 63.2% resistant to penicillin, 22.6% to methicillin, 34.8% to cephradine, 27.5% to gentamicin, and 14.4% to erythromycin; multiple resistance was common. Methicillin resistance was a feature of S. saprophyticus group strains. With isolates from catheter tips and blood cultures, a significantly higher percentage of those regarded as significant were gentamicin resistant. Apart from penicillin, antibiotic resistance was not so marked in strains of coagulase-positive staphylococci recovered over the same period.

Coagulase↗

Successive Salmonella give and Salmonella typhi infections, laboratory-acquired.

A case of laboratory-acquired typhoid fever is described. The case was complicated by a self-limiting Salmonella give gastroenteritis which may also have been laboratory-acquired and which occurred during the incubation period of the Salmonella typhi infection. The symptoms of typhoid were not sufficiently severe for the patient to seek medical attention and she was recovering from the infection when the typhoid bacillus was isolated from her stools. The mode of transmission of the S. typhi was presumed to be a laboratory infection from an unknown source. Although there was no obvious breakdown in safe laboratory techniques, the infecting dose of S. typhi is known to be small and the dangers of handling specimens which may contain this bacterium are emphasized.

Adult↗

Gross handling of impalpable breast lesion specimens.

The growing use of mammography has resulted in increasing numbers of impalpable breast lesion specimens requiring histopathological assessment. The examination of such specimens may be very time-consuming as multiple levels of many tissue blocks may need to be studied. Two grid localization methods are compared with a serial slicing method in an attempt to determine an efficient manner of identifying the impalpable lesion, and thus minimizing unnecessary processing. The serial slicing technique gives excellent radiological-pathological correlation, but is probably too complex for routine use. A simple cardboard and metal staple device gives very good localization, and is quick, easy, and cheap to use.

Breast Diseases↗

Use and usefulness of laboratory handbooks.

The aim of this study was to investigate how reference handbooks distributed by hospital laboratories are used by medical doctors, and to what extent this kind of information can influence or change doctors' work habits. We also wanted to see if books with various contents of information are valued differently by the users, and we asked for preferences for an ideal book. A questionnaire was sent to 2075 medical doctors served by five Norwegian hospital laboratories. The overall response rate was 66%, of whom 76% had received a handbook. Seventy-eight percent of respondents who stated that they had received a handbook kept it in their consulting room and 45% used it once or more weekly. The majority (89%) found the books beneficial in their everyday work. Many doctors (36%) claimed that they had changed their routines as a result of the information in the book. The way of interpreting test results was influenced most often, followed by indications for ordering laboratory tests, sample collection and specimen handling, and patient preparation. Nearly all respondents (97%) felt that handbooks of this kind are beneficial to their technical and nursing staff. The results show that handbooks distributed by medical laboratories are well received, frequently used and highly appreciated by medical doctors. Comprehensive books are rated higher than smaller books.

Education, Medical, Continuing↗

Sampling conditions for biological fluids for trace elements monitoring in hospital patients: a critical approach.

Lack of coherence in the results of trace element (TE) analyses produced by various laboratories has been such as to necessitate much needed technical improvements and more detailed control over sample preparations. For too long, except for a few praiseworthy researchers, the importance of sampling conditions has been underestimated as a source of erroneous TE results. Many of the precautionary procedures for previously studied TE collection materials have proven to be so cumbersome that they cannot be applied in a hospital setting. This report deals with sampling material and conditions from a workday standpoint; a rigorous description of the contaminating roles of ambient air and routine handling of specimens places emphasis on the practical. While some metals, including cobalt, copper, iron, and selenium, present no significant risk of contamination, others, such as aluminum, cadmium, chromium, manganese, and nickel, pose quite a significant risk. They warrant attention with respect not only to the sampling itself but also the selection of the material to be used. Drawing on our hospital experience, we propose a critical approach toward the collection of samples of biological fluids (blood, urine, dialysis fluids). Needles, catheters, tubes (open and evacuated systems), and anticoagulants on the market are likewise assessed in terms of their contaminative roles. For each one elementary rules as well as realistic choices of materials and methods for a hospital environment are proposed.

Body Fluids↗

Laboratory-acquired malaria, leishmaniasis, trypanosomiasis, and toxoplasmosis.

Because of renewed interest in parasitic diseases, increasing numbers of persons in clinical and research laboratories have the potential for exposure to parasites and therefore are at risk for acquiring parasitic infections. In this review of laboratory-acquired parasitic infections, we concentrate on protozoan diseases that frequently have been reported to be laboratory acquired: malaria, leishmaniasis, trypanosomiasis (American and African), and toxoplasmosis. These diseases can be severe, even fatal, and may be difficult to diagnose. Many laboratorians who have acquired these diseases did not recall having had an accident. Of those with recognized accidents, needlestick injuries were the most common. Laboratories should have established protocols for handling specimens that may contain viable organisms and for responding to laboratory accidents.

Animals↗

The oncologic pathology report. Quality by design.

OBJECTIVE: To review tenets of basic quality assurance and identify opportunities for quality improvement in the laboratory assessment of prognostic factors. DESIGN: College of American Pathologists Q-Probes data obtained from hundreds of American laboratories throughout the 1990s are used to illustrate laboratory performance and practice opportunities in the preanalytic, analytic, and postanalytic phases of the total testing process. RESULTS: A wide range of performance deficiencies are documented, most in the preanalytic and postanalytic phases, and contributed by individuals outside and not under the control of the laboratory. In the analytic phase, the effectiveness of checklists on the content and completeness of reported diagnostic and prognostic data for breast and colon cancers is proven. CONCLUSIONS: In the preanalytic phase, marked enhancements in quality would result from improvements in (1) patient and specimen identification, (2) adherence to patient-sampling and specimen-handling requirements, and (3) communication to the laboratory of the pertinent clinical context of the individual test. Once basic analytic quality control and laboratory quality assurance issues are resolved, a focus on standardization and enhancement of preanalytic and postanalytic communications and satisfaction of clinical expectations becomes the source of improved laboratory performance.

Humans↗

Improving quality and resource utilization through the clinical technologist.

This paper reports on the significant benefits realized during the implementation of a Clinical Technologist position in two wards of a major teaching hospital. In addition to efficient and effective handling of specimens and test results, the program results in significant savings and in enhanced patient care.

Cost Savings↗

Point of care testing: regulation and accreditation.

OBJECTIVE: To provide an overview of point of care testing (POCT) regulatory and accrediting agencies and their requirements. DATA SOURCES: Current literature and the author's experience. DATA SYNTHESIS: To develop and successfully run a POCT program that meets the federally mandated standards evaluated by accrediting agencies with Health Care Financing Administration (HCFA) status, requires that participants are aware of CLIA '88 requirements. Joint Commission on Accreditation of Healthcare Organizations (JCAHO), College of American Pathologists (CAP) and Commission on Office Laboratory Accreditation (CQLA) are the 3 principal organizations currently inspecting and accrediting POCT programs. They have included all CLIA '88 requirements on their inspection checklists. This paper covers requirements of JCAHO and CAP. In order to successfully meet the accreditation requirements of HFCA and their deeming agencies, JCAHO and CAP, these issues must be addressed: direction and supervision of the program, procedures performed, proficiency testing, quality assurance, quality control, procedure manuals, specimen handling, results reporting, reagents, calibration and standards, instrument selection and maintenance, personnel training and competence, and safety. CONCLUSION: CLIA '88 states that all testing is site neutral, therefore the same regulations apply regardless of where the test is performed. All POCT programs must apply for a CLIA certificate and choose an appropriate accreditation agency for their needs. All accreditation agencies must follow the requirements listed in the federal register; some agencies are more stringent than others.

Accreditation↗

[Difficulty in using RP, PL, and EW factors in the Japanese classification of pancreatic carcinoma].

The classification of pancreatic carcinoma by the Japan Pancreas Society reflects the prognosis of each stage better than does the TMN classification. On the other hand, there are too many factors to examine in the Japanese system, some of which are difficult to use and have low accuracy in pre- and/or intraoperative diagnosis (RP and PL), and their analysis requires various specimen handling procedures (EW). We propose that: 1) a simple, easy decision flow chart be established for ew and 2) EW/ew be designated as (-) or (+) and for EW/ew (+) cases other factors (ly, v, pl, and direct tumor invasion) be added (for example, ew (+)-pl).

Humans↗

The role of the clinical laboratory in managing chemical or biological terrorism.

BACKGROUND: Domestic and international acts of terrorism using chemicals and pathogens as weapons have recently attracted much attention because of several hoaxes and real incidents. Clinical laboratories, especially those affiliated with major trauma centers, should be prepared to respond rapidly by providing diagnostic tests for the detection and identification of specific agents, so that specific therapy and victim management can be initiated in a timely manner. As first-line responders, clinical laboratory personnel should become familiar with the various chemical or biological agents and be active participants in their local defense programs. APPROACH: We review the selected agents previously considered or used in chemical and biological warfare, outline their poisonous and pathogenic effects, describe techniques used in their identification, address some of the logistical and technical difficulties in maintaining such tests in clinical laboratories, and comment on some of the analytical issues, such as specimen handling and personal protective equipment. CONTENT: The chemical agents discussed include nerve, blistering, and pulmonary agents and cyanides. Biological agents, including anthrax and smallpox, are also discussed as examples for organisms with potential use in bioterrorism. Available therapies for each agent are outlined to assist clinical laboratory personnel in making intelligent decisions regarding implementation of diagnostic tests as a part of a comprehensive defense program. SUMMARY: As the civilian medical community prepares for biological and chemical terrorist attacks, improvement in the capabilities of clinical laboratories is essential in supporting counterterrorism programs designed to respond to such attacks. Accurate assessment of resources in clinical laboratories is important because it will provide local authorities with an alternative resource for immediate diagnostic analysis. It is, therefore, recommended that clinical laboratories identify their current resources and the extent of support they can provide, and inform the authorities of their state of readiness.

Animals↗

Ki-67 antigen detection in unstained and destained cytologic samples.

OBJECTIVE: To evaluate the effect of different tissue preservation, fixation and staining procedures on the expression of proliferation-associated Ki-67 antigen in cytologic samples to establish an easy and uniform way to handle cytologic specimens with an automated staining technique. STUDY DESIGN: Multiple touch imprints were made from eight breast tumors. The specimens were treated according to different protocols, and Ki-67 nuclear expression was compared to that in the corresponding histologic sections. RESULTS: In the unstained specimens, air drying at room temperature for up to four months or ethanol spray fixation preserved the material and offered excellent results. Processing effectively removed previous stain without additional chemical destaining. Antigen retrieval was not achieved in the previously Giemsa stained imprints and was suboptimal in those stained according to Papanicolaou. CONCLUSION: Immunocytochemical detection of Ki-67 is recommended for previously unstained cytologic specimens.

Cytological Techniques↗

Acute haematogenous anaerobic osteomyelitis in sickle cell disease. A case report and review of the literature.

Patients with sickle cell disease are more susceptible to acute anaerobic osteomyelitis due to focal gut mucosal ischaemia, translocation of bacteria, and seeding in infarcted bone marrow. Modulation of the immune system is also present. The isolation of anaerobic organisms requires a high index of suspicion, correct specimen collection procedures and meticulous specimen handling. Bacteroides is the predominant organism isolated. Intra-osseous gas in the bone may be seen within four days and radiographs are therefore useful earlier than with aerobic osteomyelitis. Surgical débridement and intravenous antibiotics are the mainstay of treatment with the erythrocyte sedimentation rate being relied on heavily to guide conversion to oral antibiotics. Coexistence of septic arthritis is more common with anaerobic osteomyelitis.

Acute Disease↗

A novel filtration-based processing method of liquid cytology specimens for human papillomavirus DNA testing by hybrid capture II.

We evaluated a more efficient method of processing liquid-based cervical cytology specimens for human papillomavirus (HPV) DNA testing by Hybrid Capture II (HCII). Aliquots were made from 701 specimens in the following sequence: 4.0, 2.0, 1.0, 0.5, and 1.5 mL. The 4.0-mL aliquot was processed by the standard method (STP), and half of the processed material was tested by HCII. Other aliquots were processed with a new, filtration-based processing method (NPM). The 2.0-mL NPM aliquot had HCII test performance most similar to the STP, ie, similar HCII positivity (P = .4) and good test agreement (kappa = 0.85, 95% confidence interval [CI], 0.80-0.89). The 194 cytologic negatives had greater positivity by STP (P = .04) compared with the 2.0-mL aliquot processed by NPM; between-method agreement was modest (kappa = 0.54, 95% CI, 0.36-0.72). A lower positive cut point for the 2.0-mL NPM aliquot partially abrogated this minor difference. In 241 specimens diagnosed as low-grade and 31 as high-grade squamous intraepithelial lesions, there were no significant differences in HPVpositivity (>85% and 90%, respectively) between STP and NPM. NPM reduces specimen handling and decreases total testing time by approximately 33% without significant losses in HCII test performance.

DNA, Viral↗

Procedures in primary care dermatology.

BACKGROUND: General practitioners can provide a range of diagnostic and treatment procedures for patients with dermatological problems. OBJECTIVE: This article reviews the use of Wood's light, diascopy, skin scrapings for microscopy, skin biopsy, cauterisation, doppler measurement of ankle-brachial pressure index, and multi-layer compression bandages for venous ulcers. DISCUSSION: Adequate training and proper equipment is essential for commonly neglected areas including informed written consent, good communication with the histopathologist, a system for handling specimens and reports, action plans for laboratory results, and adequate training and protection for auxiliary staff.

Bandages↗

Adsorption with a soluble E. coli antigen fraction improves the specificity of ELISA tests for Lyme disease.

We reported that preadsorption of patient serum with heat killed E. coli increased the specificity of ELISA for antibodies to Borrelia burgdorferi. That procedure required extra specimen handling and a preincubation. We report the use of a soluble E. coli antigen fraction that is included in serum diluent, eliminating additional steps. Sera from 220 individuals were tested for antibodies to B. burgdorferi. Twenty sera were obtained from patients with Lyme disease and 200 sera were from a population that included healthy controls and patients with different inflammatory conditions (viral infections and various rheumatic disorders). Testing was performed using either a standard serum diluent or one containing soluble E. coli antigen fraction. Results demonstrate that inclusion of soluble E. coli antigen fraction in serum diluent increased assay specificity from 88% for the standard protocol to 98%, with no change in test sensitivity.

Antigens, Viral↗