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Brucellosis in laboratory workers at a Saudi Arabian hospital.

BACKGROUND: Saudi Arabia is hyperendemic for brucellosis, with more than 8000 cases reported each year to public health authorities. During 1998, brucellosis ranked as the No. 1 reportable communicable disease (22.5%) in Saudi Arabian National Guard communities. King Fahad Hospital is the major referral center for National Guard personnel in the nation's central region. METHODS AND RESULTS: From 1991 to 2000, brucellosis developed in 7 expatriate hospital employees. Six employees were bacteriology technologists, and one was a pathologist. Each had a clinical syndrome compatible with brucellosis (headache, fever, rigors, sweats, and myalgias) plus elevated Brucella sp serum agglutinin titers > or = 1:1280; one patient also had positive blood cultures. All patients responded to anti-Brucella therapy. Two patients had relapses, and complications occurred in four patients (septic endophlebitis of the leg, infected prosthesis, epididymoorchitis, and lumbar spondylitis). In all these employees except the pathologist, the infection was associated with processing Brucella sp cultures. CONCLUSION: Despite the enforcement of stringent infection control measures including the use of a class II biosafety hood in the laboratory, the problem of nosocomial brucellosis persists because of the large number of infected specimens handled by the laboratory (17,500 specimens per year). Ultimately, risk reduction depends on efforts to reduce disease endemicity in the country. In the meantime, conversion of the laboratory to biosafety level 3 is under way.

Animals↗

Allergic fungal sinusitis (AFS)--earlier diagnosis and management.

AIM: To evaluate the criteria for diagnosing allergic fungal sinusitis and to maintain permanent drainage and ventilation, while preserving the integrity of mucosa. METHODS: This is a prospective study of 251 patients with chronic rhinosinusitis with or without polyposis, of whom 199 were treated surgically. Mucus sample collection, nasal secretion culture, surgical specimen handling and histological evaluation of surgical specimens are described. The management included wide local endoscopic sinus debridement, adequate sinus aeration, post-operative use of steroids and antifungal therapy. RESULTS: Fungal cultures of nasal secretions were positive in 201 (80.01 per cent) of 251 patients. Of the 199 surgical cases, fungal elements were found in 156 histological specimens (62.1 per cent). Allergic mucin was found in 182 patients (91.45 per cent). Nasal obstruction and proptosis were the commonest presentations. All pre-operative versus post-operative changes in AFS-associated complaints reached statistical significance of p < 0.001. The ethmoid sinus was commonly involved with adjacent lamina papyracea exhibiting demineralization in 26.6 per cent of cases. Intracranial extension was seen in 15 cases. Recurrence was noted in 11 cases. CONCLUSION: Comprehensive treatment with endoscopic sinus surgery, steroids and antifungal therapy is needed. AFS is readily recurrent. Long-term follow up is important.

Adolescent↗

Specimen collection and storage for diagnostic molecular pathology investigation.

The success of the newest discipline in the diagnostic clinical pathology laboratory, molecular pathology, is dependent on proper collection and storage of both the original and processed (nucleic acid) specimen. This issue will grow in importance as test volumes increase in the diagnostic molecular pathology laboratory. This review is a distillation of a literature review by the Patient Preparation and Specimen Handling Committee of the College of American Pathologists. It describes specific collection, storage, and anticoagulant or preservative requirements based on the diagnostic molecular technique and/or specimen type used for analysis. This review serves as a guide for clinical laboratories interested in appropriate collection and storage of specimens to be used in nucleic acid-based analysis.

DNA↗

Tumor cell dissemination during laparoscopy: prevention and therapeutic opportunities.

Port-site recurrences (PSRs) are abdominal wall recurrences that occur in the subcutaneous tissue within a trocar site after cancer laparoscopy and are not associated with peritoneal carcinomatosis. In order to develop PSRs, viable tumor cells must be liberated from the primary tumor, be transported to a wound, and find there a favorable environment for growth. The short clinical delay in the occurrence of PSRs and their size suggest massive cell seeding into the abdominal wall. Traumatic handling of the tumor, slipping of trocars, liquid projection, as well as poor extraction techniques can all cause implantation of malignant cells into the subcutaneous tissue. Such contact can also occur postoperatively if the trocar channels remain open. Some histologies (e.g. gallbladder adenocarcinoma), the presence of ascites and advanced tumor stage are risk factors for PSRs. Further conditions--including the use of gas--might also play a limited role. The first preventive measure is the correct indication for a laparoscopic approach. Several techniques have been demonstrated to prevent PSRs in the animal model: (a) fixation of trocars to the abdominal wall; (b) prevention of leakage; (c) careful specimen handling; (d) reducing trauma to the abdominal wall; (e) specimen isolation before extraction from the abdominal cavity; (f) trocar-site irrigation with a cytotoxic solution, and (g) closure of peritoneum. Further innovative therapies are currently under investigation. In the clinical setting, correct indication, surgical expertise and application of prophylactic measures seem to be the best way to prevent the occurrence of PSRs.

Abdominal Wall↗

Use of bar code labels on collection tubes for specimen management in the clinical laboratory.

A new generation in specimen handling has arrived with the introduction of bar code readers on medical laboratory equipment. The incorporation of this technology into laboratory information systems offers a streamlining of specimen workflow never before achievable in a laboratory environment. The use of evacuated collection tubes as the primary sampling container on a random-access chemistry analyzer interfaced to a laboratory information system creates a very simplified sampling/analysis system with tremendous advantages. There are significant labor savings, superior service to clinicians, and reduced chances for clerical error.

Computers↗

Guidelines for performing single-platform absolute CD4+ T-cell determinations with CD45 gating for persons infected with human immunodeficiency virus. Centers for Disease Control and Prevention.

These guidelines were developed by CDC for laboratorians who perform immunophenotyping for detection and enumeration of CD4+ T-cells and other lymphocyte subsets in persons infected with human immunodeficiency virus (HIV). The guidelines describe single-platform technology (SPT), a process in which absolute counts of lymphocyte subsets are measured from a single tube by a single instrument. SPT incorporates internal calibrator beads of known quantity in the analysis of specimens by three- or four-color flow cytometry. With CD45 gating, the relative numbers of beads and lymphocyte subsets are enumerated, and their absolute numbers and percentage values are calculated. This report supplements previous recommendations published in 1997 (CDC. 1997 revised guidelines for performing CD4+ T-cell determinations in persons infected with human immunodeficiency virus [HIV]. MMWR 1997;46[No. RR-2]) that describe dual-platform technology, a method in which absolute counts are derived from measurements obtained from two instruments--a flow cytometer and hematology analyzer. The new recommendations address concerns specific to the implementation of SPT as well as other general topics such as laboratory safety and specimen handling.

CD4 Lymphocyte Count↗

Pathologic analysis of routine tonsillectomy and adenoidectomy specimens.

OBJECTIVES: Recent literature has suggested that histopathologic analysis of routine tonsillectomy and adenoidectomy (T&A) specimens may be unnecessary. This study investigates T&A specimen handling practices in the United States between 1989 and 1999. METHODS: Surveys were sent to 4715 members of the American Academy of Otolaryngology. Surveys assessed practice type, pathologic processing practices (full, gross, no pathology), and reasons for change. The authors also performed a retrospective analysis of 1583 pediatric T&A specimens for evidence of occult malignancy. RESULTS: Practice types were 80% private, 12% academic, 6% salaried, and 2% military. Chi squared analysis revealed a significant increase (P < 0.001) in respondents ordering "gross only" and "no pathology." The retrospective analysis found no occult malignancies. CONCLUSIONS: There is a statistically significant increase in the number of otolaryngologists sending routine T&A specimens for "gross only" and "no pathology." There was no correlation between the type of practice and changes in pathologic analysis performed.

Adenoidectomy↗

Optimal fecal assessment.

Fecal testing is a common component of most gastrointestinal work-ups. A multitude of diagnostic techniques are available for identifying parasites and pathogens, or showing abnormalities of flora. Optimal fecal assessment involves careful formulation of a differential list based on signalment, history, and clinical signs. Tests should be selected and interpreted based on their relative sensitivity and specificity for specific conditions. It is essential to use effective testing methods for the etiologies of concern. This article reviews the plethora of diagnostic techniques available for fecal assessment. Indications, limitations, and issues of specimen handling for each technique are discussed. The optimal approach to the diagnosis of some common parasites, pathogens, abnormalities of flora, and metabolic conditions are covered.

Animals↗

DNA probes for the detection of mycoplasmas in genital specimens.

The utility of whole-genomic DNA probes for the detection of infections by genital mycoplasmas was investigated in 220 men attending a sexually transmitted diseases clinic. In 144 patients, probe results were compared with quantitative culture results. The prevalence of Mycoplasma hominis was 11% by culture, whereas the prevalence of ureaplasmas was 38%. The M. hominis DNA probe detected 9 of 16 M. hominis culture-positive specimens and 2 of 128 culture-negative specimens. The Ureaplasma urealyticum DNA probe detected 36 of 57 U. urealyticum culture-positive specimens and 18 of 87 culture-negative specimens. Most of the probe-negative culture-positive specimens had colony counts of less than 10(3) organisms/ml of specimen. The DNA probe does not require viable organisms, and the probe-positive, culture-negative specimens suggest that false-negative cultures occurred, perhaps due to specimen handling or insensitivity of culture methods for some strains of mycoplasmas.

Bacteriological Techniques↗

Vision assisted robotics and tape technology in the life-science laboratory: applications to genome analysis.

Recent proposals to analyze a number of major genomes have created a need for methods of rapidly manipulating and assaying very large numbers of specimens containing DNA fragments. Systems for sampling semi-solid biological material from mixed disordered arrays are required. These samples need to be sorted in an ordered format and stored in minimum space at known locations. It should be possible to recall them quickly from stock, individually or collectively, for duplication or re-ordering into new arrays for appropriate analysis. This article reviews a range of problems associated with high speed multiple specimen handling and assay in the molecular biology laboratory and outlines the solutions currently available to address these difficulties. Designs for automatic systems to manipulate biosamples and analyze DNA are presented.

Biotechnology↗

Pathology of transitional cell carcinoma of the bladder and its clinical implications.

Transitional cell carcinomas are divided into superficial and muscle-invasive tumors. Most of them are superficial tumors, and approximately 15-20% are muscle-invasive carcinomas. Pathologists play a significant role in diagnosing bladder tumors and in reporting features important for determining prognosis. We will review the cytologic and histopathologic features that help determine prognosis, including depth of invasion, tumor grade, multicentricity, tumor size, and the presence of vascular/lymphatic invasion, blood group antigen expression, proliferative indices, and molecular markers. Brief mention will be made of specimen handling, interpretation, reporting, and histologic variants of transitional cell carcinoma.

ABO Blood-Group System↗

The collection and handling of the blood alcohol specimen.

Proper collection, handling, and storage of the blood alcohol specimen are essential in medicolegal cases involving the question of sobriety. A standard operating procedure is necessary to ensure maximum reliability. Comments are offered on the advantages of using blood specimens in preference to urine or tissue specimens. The use of a conversion factor to obtain a calculated "presumed blood level" can be dangerous. Cautions and suggestions are offered regarding how and from where the blood should be obtained from a living person and during an autopsy. There are certain time limitations for storage of these blood-alcohol specimens. Each laboratory must establish its own limits for reliable storage, given the conditions in that laboratory. Unexpected and confusing results can lead to an erroneous interpretation if history, circumstances, type of injury, and survival time are not all carefully considered. Several possibilities for error in judgment are discussed.

Ethanol↗

Molecular diagnostics. The polymerase chain reaction and its use in the diagnosis of Chlamydia trachomatis and Neisseria gonorrhoeae.

The diagnosis of Chlamydia trachomatis and Neisseria gonorrhoeae was performed using classical microbiological techniques of chlamydial cell culture and agar isolation respectively in patients studied in Pittsburgh. The polymerase chain reaction (PCR) was compared to standard procedures used for diagnosing these sexually transmitted organisms. Statistically significant differences were observed in molecular diagnostics versus classical isolation techniques. Numerous specimen handling problems were identified in the handling of Neisseria. The enhanced ability to diagnose these sexually transmitted organisms is discussed in relation to the Human Immunodeficiency Virus (HIV).

Cervix Uteri↗

The dos and don'ts of an oral mucosal biopsy performed by the general dentist.

The recognition of a pathologic entity is only the first step in the chain of events which must be performed for accurate diagnoses and appropriate patient management. Other steps include selection of the type of biopsy to be performed; proper biopsy technique, instrumentation and specimen handling; appropriate fixation; detailed documentation; and postoperative patient instructions. If the clinician performs each step carefully and correctly, the patient is ensured of receiving an accurate diagnosis in the shortest possible period of time.

Biopsy↗

Urine markers as possible tools for prostate cancer screening: review of performance characteristics and practicality.

BACKGROUND: In recent years, an increasing number of urine-based tests have been proposed as potential screening tests for prostate cancer. The goal of this review was to summarize the current status of evidence regarding performance characteristics of the proposed tests and their practicality under screening conditions. METHOD: Relevant articles published up to and including May 2005 were identified in the PubMed database. At least 10 cases and 10 controls had to be analyzed for a study to be included in the review. Data concerning the study population, performance characteristics, and the collection and processing of urine samples were extracted from the reviewed articles. RESULTS: In all, 34 retrospective studies evaluating 21 different markers complied with the inclusion criteria. Most of the studies were rather small and included heterogeneous clinical study populations. Promising results were reported for a few markers in single studies, but they have often not been replicated in subsequent larger studies. Some of the more promising results were obtained with 24-h urines or with specimen-handling procedures that might be difficult to perform under screening conditions. CONCLUSIONS: Larger studies with a prospective design are required to confirm promising findings regarding performance characteristics of some novel markers recently reported in mostly small studies. Future studies should also pay particular attention to the practicality of the markers under screening conditions.

Biomarkers, Tumor↗

Stereotactic core-needle biopsy of the breast: a report of the Joint Task Force of the American College of Radiology, American College of Surgeons, and College of American Pathologists.

A national task force consisting of members from the American College of Radiology, the American College of Surgeons, and the College of American Pathologists examined the issues surrounding stereotactic core-needle biopsy for occult breast lesions. Their report includes indications and contraindications, informed consent, specimen handling, and management of indeterminate, atypical, or discordant lesions.

Biopsy, Needle↗

A practical strategy for diagnosis of urinary tract infections.

Because urine specimens are not all identical, clinical microbiology laboratories should tailor specimen handling and processing to the specific clinical setting. To quantify uropathogens requires specific ranges that depend on the clinical presentation of a patient and the collection method. A single quantitative breakpoint, for example, the classic "100,000 CFU/mL" cannot be applied to patients with AUS or with indwelling catheters. This article has attempted to review rational adjustments in the traditional urine culture protocol to fit both the clinical setting and the mode of collection.

Catheters, Indwelling↗