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Handling of clinical tissue specimens for molecular profiling studies.

The relationship between gene expression profiles and cellular phenotypes is an important aspect of functional genomics. Clinical tissue specimens will play a vital role in this effort. The usefulness of tissue for molecular profiling is significantly influenced by the manner of specimen handling. Crucial components of this process include the optimization of the methods of tissue fixation and embedding, not only to obtain excellent histological detail, but also to promote the elucidation of the gene and protein expression profiles. In this article, we describe handling of clinical specimens using whole-mount prostate as an example, the use of new high-throughput techniques that allow molecular profiling analysis and the use of a web-based 3-dimensional model to combine these data to make it available to clinicians and the research community. Complete protocols and additional discussion are available on the website, http://cgap-mf.nih.gov.

Cryopreservation↗

Urinary retinol binding protein: stability and pre-analytical handling of specimens for its measurement.

The stability of retinol binding protein (RBP) in urine has been reassessed and was found to be pH-dependent, diminishing as urinary pH decreased. When urines were incubated at 4 degrees C for 1 week, statistically significant losses of RBP were observed at pH values less than seven, varying in degree in different urines. At 37 degrees C some urine specimens showed minor losses when incubated for periods up to 4 h. This was more marked at pH 5 than pH 7 with maximum losses of 26% and 12%, respectively. When adjusted to pH 7, urines showed no significant loss of RBP when stored at 4 degrees C for up to 2 weeks; at -20 degrees C no significant change in RBP concentration was found for at least 14 weeks. To minimize losses of RBP it is recommended that urine pH is adjusted to above seven immediately after voiding.

Enzyme-Linked Immunosorbent Assay↗

[Safety measures for handling laboratory specimen and patients with Creutzfeldt-Jakob disease].

Creutzfeldt-Jakob disease (CJD) is a transmissible subacute spongiform encephalopathy that invariably leads to death. The presumed causative agent, the prion protein, is highly resistant to inactivation and has a long incubation period. Physical contact with CJD patients (as in clinical care) entails no risk of transmission. During procedures such as lumbar puncture where contact with infected material is possible, precautions are necessary. Precautions are: the use of gloves, maximal protection of people who come in contact with contaminated tissue (e.g. pathologist and histological laboratory worker) and transportation of samples in a closed and labelled container. For laboratory research the tissue must be submerged in 92-98% formic acid for 1 hour. All used materials and instruments must be decontaminated properly, using for instance NaOH, NaClO, guanidine thiocyanate or steam autoclaving. If adequate precautions are taken contact with contaminated materials can be safe.

Creutzfeldt-Jakob Syndrome↗

A container for handling small specimens during preparation and examination in the scanning electron microscope (SEM).

The difficulties of handling small specimens of less than 1 mm3 are considerably reduced when the specimens are placed in a small container designed to be compatible with the dimensions of the preparatory instrumentation and of the microscope. The container described is unaffected by solvents used in the preparation of specimens for scanning electron microscopy.

Chlamydomonas↗

Handling and reporting of biopsy and surgical specimens of testicular cancer.

This paper is the result of a meeting of the European Association of Pathologists, Uropathology Division in Florence 2003. The aims of this meeting were to establish: guidelines for specimen handling by urologists and minimum requirements for data accompanying testicular specimens submitted to pathologists; a consensus on techniques for processing specimens by pathologists; the essential information required from pathology reports; areas where our standard practice is traditional rather than evidence based and where further studies are required. The general aims of histopathology are to give or confirm a diagnosis; assess established prognostic markers; identify changes associated with treatment; provide information for audit (i.e. imaging, urology and pathology) and maintain a permanent record (slides/blocks).

Biopsy↗

Handling and pathology reporting of renal tumor specimens.

The gold standard in the treatment of renal tumors is radical or partial nephrectomy. The surgical specimen handling is important since the pathologic features result in clinico-pathologic knowledge that determines prognosis, additional treatment and scientific studies. The correct handling of the specimen by urologists and pathologists becomes very basic in order to enable retrieval of a maximum of information. This protocol aims at standardization of the minimal criteria for handling, cellular subtyping, grading, staging and margin evaluation that must allow the comparison among the different scientific groups.

Humans↗

Qualitative and quantitative determinations of acid phosphatase activity in vaginal washings.

In the absence of spermatozoa, acid phosphatase activity (ACP) is an indicator of seminal fluid in cases of sexual assault. Both qualitative and quantitative ACP methods are employed by medical examiners. A large series of cases in which a qualitative test was employed is reported. A method for obtaining and handling specimens by vaginal washing was developed. This method provides suitable specimens for the examination for spermatozoa and the determination of ACP. Specimens obtained from 41 women provided examples of endogenous vaginal ACP and intravaginal seminal ACP. These specimens were used to evaluate qualitative and quantitative ACP methods for the detection of seminal ACP. Tartrate inhibition was not useful for separating endogenous vaginal and seminal ACP activities. The advantages of the method developed for specimen acquisition and of quantitative assays for ACP are discussed.

Acid Phosphatase↗

Suspected cutaneous anthrax in a laboratory worker--Texas, 2002.

On March 6, 2002, CDC's National Institute for Occupational Safety and Health (NIOSH) received a request for a health hazard evaluation from the director of Laboratory A to assist in the evaluation of a worker who had been diagnosed with cutaneous anthrax. Laboratory A, a provisionally approved Laboratory Response Network level B laboratory, had been processing environmental samples for Bacillus anthracis in support of CDC investigations of the bioterrorist attacks in the United States during fall 2001. Since March 7, CDC has interviewed the ill laboratory worker and other workers at the laboratory and conducted environmental assessments of the workplace. This report summarizes the epidemiologic and environmental investigation of this case, which indicates that the likely source of exposure was the surface of vials containing B. anthracis isolates that the worker placed in a freezer on March 1. Laboratory workers handling specimens of B. anthracis should follow recommended procedures to minimize the risk of B. anthracis transmission and anthrax.

Anthrax↗

The effect of saliva specimen collection, handling and storage protocols on hepatitis C virus (HCV) RNA detection by PCR.

OBJECTIVES: Commercial assays can now be adapted to detect salivary anti-hepatitis C virus (HCV) antibodies, increasing the potential of saliva as a non-invasive diagnostic specimen suited to surveillance and epidemiological studies. However, current diagnostic algorithms involve confirmation of HCV infection by RT-PCR. Manipulation and storage conditions of serum can influence the stability of viral RNA. This study examined whether varying specimen collection, handling and storage protocols also affected subsequent HCV RNA detection by RT-PCR applied to saliva specimens. METHODS: Whole unstimulated saliva, together with saliva samples collected in two commercially available devices (Salivette and Omnisal) were obtained from 50 HCV seropositive intravenous drug users. The specimens were subjected to a number of handling and storage conditions, including heat treatment and prolonged storage, then examined for HCV RNA by RT-PCR using primers derived from the 5' non-coding region (5'NCR). RESULTS: HCV RNA was detected in saliva samples from 25 (50%) of the patients. No single collection device or handling procedure identified all the subjects with HCV RNA positive saliva though whole saliva yielded the greatest number of positive results. CONCLUSIONS: Collection and processing of saliva specimens for RT-PCR analysis is complex. At present, detection of salivary HCV RNA by PCR is not sufficiently sensitive for use as a diagnostic tool in epidemiological studies.

Adult↗