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PubMed2005

Effects of delayed microbial analysis of dental unit water line specimens.

PURPOSE: Monitoring microbial concentrations in water emitted from dental unit water lines (DUWL) is an important safety procedure. Improper handling of test water specimens could give incorrect results. Thus, the objective of this study was to measure the effects delayed culturing might have on DUWL specimens. METHODS: First, 100 mL water specimens were obtained from 10 different handpiece service lines within the School. All units had independent water systems, used DI (deionized water) water and were routinely cleaned using an alkaline peroxide based product. Two specimens of 10 mL were removed from the bottles and placed into individual sterile conical tubes. One set of tubes was processed immediately. 0.05 mL of sterile 1.0% (w/v) sodium thiosulfate solution was added to undiluted and diluted (1:10 and 1:100 with sterile DI water) specimens. After mixing, specimens were spiral plated onto duplicate R2A plates and incubated at 21degrees C for 7 days. Colonies were then counted and the cfu/mL of each original specimen determined. Another set of tubes was placed into a shipping envelope and mailed out to the School. Upon receipt, the tubes were processed as described above. The remaining 80 mL of water in the collection bottles were divided equally into new sterile tubes. One tube was left at 21 degrees C, while the other was placed into a 37 degrees C incubator. Aliquots were processed immediately and then after 1, 3 and 7 days. Next, 30 mL water specimens were obtained from 15 handpiece service lines in three outside clinics. All units had independent water systems, used DI water and were routinely cleaned with an alkaline peroxide-based product. Specimens were then divided equally into three sterile conical tubes. One of the tubes was transported (at 4 degrees C) to the laboratory and immediately processed as described. At the collection site, the second tube was placed into a padded envelope and mailed back to the School. The third tube was returned by overnight delivery using a Cool Pack type container. Upon receipt, all the tubes were processed as described previously. RESULTS: The 10 handpiece waterline specimens processed immediately ranged from 0 to 1000 cfu/mL. Holding specimens at 21 degrees C produced radically higher bacterial counts (1540-866,000 cfu/mL) in water from 90% of the handpieces. Holding at 37 degrees C produced unacceptably high bacterial counts in only 50% of the handpiece specimens. Mailed specimens were cultured 5 days after collection and water of unacceptable quality water was noted in 70% of the specimens. In another experimental set, mailed specimens arrived after 72 hours and were an average of 20 degrees C. Express sent specimens came the next morning at an average temperature of 4.5 degrees C. Only one waterline specimen processed immediately contained more than 500 cfu/mL. In contrast, 80% of specimens returned by post at ambient temperature had unacceptably high bacterial counts (780-376,000 cfu/mL). Express sent specimens produced the same results as those processed immediately.

Bacteria, Aerobic↗

PubMed2006

Evaluation of the Dako IDEIA norovirus EIA assay for detection of norovirus using faecal specimens from Australian gastroenteritis outbreaks.

AIMS: New techniques for detection of norovirus, a major cause of gastroenteritis, require ongoing evaluation. The aim of this study was to use material from gastroenteritis outbreaks in Victoria, Australia, to evaluate the sensitivity and specificity of the Dako IDEIA norovirus EIA assay, using both photometric and visual analysis. METHODS: A total of 130 faecal specimens from 41 gastroenteritis outbreaks were tested for norovirus by electron microscopy (EM), a two-round multiplex reverse transcription-polymerase chain reaction (RT-PCR) method and the IDEIA norovirus assay. All specimens with sufficient amplified product were sequenced to determine their norovirus genotype. In addition, six well-established RT-PCR protocols were used to test four EIA-positive, multiplex RT-PCR/EM-negative specimens. Also, a range of RT-PCR protocols was used to test a specimen positive for GII only by the multiplex RT-PCR but positive for GI and GII by the EIA. The effect of multiple freezing-thawing cycles on EIA positivity was tested on seven additional specimens. A further seven specimens, known to contain the gastroenteritis viruses sapovirus, adenovirus, astrovirus and rotavirus were also tested by the IDEIA norovirus assay. RESULTS: The IDEIA norovirus assay gave a single-specimen sensitivity and specificity of 66% and 85%, respectively (visual analysis compared with the multiplex RT-PCR), 63% and 88% (photometric analysis compared with the multiplex RT-PCR), 65% and 87% (visual analysis compared with the multiplex RT-PCR and/or EM) and 62% and 90% (photometric analysis compared with the multiplex RT-PCR and/or EM). None of the four EIA-positive specimens negative by the multiplex RT-PCR and/or EM was positive by any of the six alternative RT-PCR protocols. The specimen positive for GI and GII by EIA but for GII only by the multiplex RT-PCR was not positive for GI by any of the alternative RT-PCR protocols. A minimum of three specimens per outbreak had to be tested by the EIA to ensure that norovirus-positive outbreaks (multiplex RT-PCR and/or EM) were classified as positive for norovirus by the IDEIA norovirus assay (visual or photometric analysis). However, one specimen from a norovirus-negative outbreak (multiplex RT-PCR and/or EM) for which four specimens were provided was positive for norovirus by the IDEIA norovirus assay. Seven norovirus genotypes were identified by open reading frame 1 sequencing analysis and specimens from all seven norovirus genotypes (as well as an EM-positive/multiplex RT-PCR-negative specimen) were detected by the IDEIA norovirus assay by both visual and photometric analysis. Repeated freezing-thawing cycles (up to six) for faecal specimens did not reduce the sensitivity of the EIA assay but could render an EIA-negative specimen EIA-positive. The specimens positive for sapovirus, adenovirus, astrovirus and rotavirus were EIA-negative. CONCLUSIONS: The IDEIA norovirus assay lacks the sensitivity and specificity to ascribe a particular result to a particular specimen, but could be useful for detecting norovirus in a gastroenteritis outbreak where specimens are plentiful, although it is difficult to avoid a risk of false positives. Since visual analysis can be used for result assessment almost as reliably as photometric analysis, the test kit would be useful for laboratories lacking specialist equipment such as a photometric microplate reader.

Caliciviridae Infections↗

PubMed1998

Transbronchial lung biopsy: can specimen quality be predicted at the time of biopsy?

STUDY OBJECTIVES: To determine the bronchoscopist's ability to predict specimen quality at the time of transbronchial biopsy and to determine the influence of biopsy specimen size and alveolar content on diagnostic value. DESIGN: Prospective, blinded, observational analysis. SETTING: Tertiary care academic hospital-based pulmonary practice. PATIENTS: Forty-three adult patients who underwent transbronchial lung biopsy. INTERVENTIONS: Each of 170 biopsy specimens was rated as to likelihood of containing diagnostic tissue, size and ability to float, tissue types present, number of alveoli, and pathologic diagnosis. RESULTS: Fifteen percent of biopsy specimens were small and 40% were large. Seventy-six percent of specimens floated; 61.8% of the 170 biopsy specimens contained abnormal lung tissue; and 14.7% of individual specimens were diagnostic. Fifty-two percent of specimens contained >20 alveoli. Larger biopsy specimens were more likely to contain diagnostic tissue (r=0.29, p=0.001). Cup forceps retrieved smaller pieces of tissue (p=0.007) and were less likely to obtain diagnostic tissue (p=0.06). Physician ratings of specimen quality (mean+/-SD) did not differ between specimens containing normal and abnormal tissue (5.98+/-2.3 vs 5.46+/-5.5; p=0.24) or between specimens containing diagnostic vs nondiagnostic tissue (5.56+/-2.5 vs 6.25+/-2.1; p=0.14). Specimens that floated were no more likely to be diagnostic or abnormal than specimens that sank (p<0.05). Diagnosis when established was made by the first biopsy specimen in 53.3% and the second in 33.3% CONCLUSIONS: Physician estimate of biopsy specimen quality and the float sign are not helpful in predicting that the biopsy specimen contains abnormal or diagnostic tissue. Diagnostic biopsy specimen will likely be obtained if the size of the specimen fills the forceps, 2 to 4 biopsies are performed, and toothed forceps are used.

Adult↗

PubMed1997

Validity of clinic biopsy specimens in classifying histopathologic characteristics of recurrent nasopharyngeal carcinoma.

OBJECTIVE: To evaluate nasopharyngeal carcinoma resection specimens for heterogeneity of histologic patterns to determine if preoperative histologic characteristics of the clinic biopsy specimen are representative of the entire lesion. The null hypothesis is that clinic biopsy specimens are not necessarily representative. DESIGN: Preoperative clinic biopsy specimens were measured to calculate their average size. Resection specimens were then sectioned and evaluated in increments corresponding to this size. Each of these increments was then histologically classified according to the World Health Organization (WHO) criteria. This classification of the preoperative biopsy specimens was compared with that of the resection specimen as a whole. SETTING: University referral center. PATIENTS: Twenty-six consecutive patients with recurrent nasopharyngeal carcinoma who underwent surgical resection. Radiation therapy failed in all patients. MAIN OUTCOME MEASURE: The presence or absence of WHO histologic heterogeneity in the nasopharyngectomy specimen was recorded. Disparity between preoperative clinic biopsy and resection specimens was recorded. RESULTS: The mean clinic biopsy specimen size was 13.9 mm2 or less than 1% of the available surface area of the nasopharynx. Of 26 resection specimens classified in 5 increments of this size, 15 (57.7%) were a single WHO type, and 11 (42.3%) were found to be mixtures of WHO types I, II, and III. Of 16 cases with preoperative biopsy specimens available, 4 (25%) were a different WHO classification than their corresponding resection specimen. CONCLUSIONS: Most clinic biopsy specimens were representative of their corresponding tumor resection specimens in their entirety; however, tumor heterogeneity is such that some biopsy specimens will not be representative. This finding may interfere with WHO classification data determined on the basis of clinic biopsy specimens and hence confound any meaningful data on treatment outcomes. It is recommended then that multiple nasopharyngeal biopsy specimens be obtained from disparate areas of the lesion and each subjected to independent histopathologic review.

Adult↗

PubMed1999

Telomerase activity: comparison between fine-needle aspiration and biopsy specimens for the detection of tumor cells.

BACKGROUND: The purpose of the current study was to determine telomerase activity as a sensitive biomarker for the detection of malignant cells in fine-needle aspiration (FNA) specimens. METHODS: FNA specimens with parallel samples of fresh tumor tissue were obtained from surgical specimens after surgical excision. Using a polymerase chain reaction-based telomeric repeat amplification protocol (TRAP) assay, telomerase activity was determined systematically in FNA specimens (n = 21) and corresponding available tissue biopsy specimens (n = 16) containing malignant cells. In addition to a case of myelolipoma, normal counterparts for 3 of 16 cancer cases, including both biopsy and FNA specimens, also were available for the determination of telomerase activity. RESULTS: Telomerase activity was observed in 14 of 16 of the FNA specimens (88%) and 15 of 16 of the corresponding biopsy specimens (94%). Telomerase activity was detected in both the biopsy specimen and the corresponding FNA specimen, with one exception (a case of mucinous adenocarcinoma of the cecum). In contrast, specimens from three normal tissue biopsies and FNA specimens of normal tissue adjacent to the malignant lesions, as well as the myelolipoma, exhibited no telomerase activity. It is interesting to note that both tissue biopsy specimens and FNA specimens from a patient with high grade sarcoma were negative for telomerase activity. The examination of hematoxylin and eosin-stained adjacent tissue biopsy sections or FNA smears revealed similar low populations of lymphocytes, including those cases that were negative for telomerase activity. There was agreement in the detection of telomerase activity between tissue biopsies and their corresponding FNA specimens in 15 of the 16 patients, indicating a 94% concordance rate (95% confidence interval, 70%, 98%). CONCLUSIONS: The results of the current study clearly suggest that the telomerase activity in FNA specimens was comparable to that of their corresponding biopsy specimens, and that this activity was associated with the presence of malignant cells. The TRAP assay has potential for use in the detection of malignant cells in FNA specimens, particularly cases in which the cytology is not characteristically malignant and/or is present in insufficient numbers. Cancer (Cancer Cytopathol)

Adolescent↗

PubMed2001

Effect of endocervical specimen adequacy on ligase chain reaction detection of Chlamydia trachomatis.

Several studies have demonstrated that the sensitivity of a commercially available PCR test for the detection of Chlamydia trachomatis (Roche Diagnostics) is affected by the cellular quality of the endocervical swab specimens. The cellular adequacies of 1,633 female endocervical swab specimens were assessed and compared with the results of C. trachomatis detection obtained by ligase chain reaction (LCR; Abbott Laboratories). Specimen adequacy studies and LCR were performed with samples from the same swab, after demonstration of the stability of human epithelial cells in LCR transport medium. Prior to heat treatment of the swab specimen, an aliquot was removed and cytocentrifuged onto a slide. Cell spots were stained and examined at x 400 magnification for endocervical (columnar epithelial or metaplastic) cells and erythrocytes. The overall rate of positivity of the LCR was 6.5% (106 of 1,633 specimens) with pooled specimens (pools of 4 specimens each; reduced cutoff). Of the 1,633 specimens examined, 655 (40.1%) were found to contain one or more endocervical cells. The rate of positivity for C. trachomatis was 10.8% (71 of 655 specimens) among specimens containing endocervical cells, whereas it was 3.6% (35 of 978 specimens) among specimens lacking endocervical cells (P < 0.0001). There was no linear trend between the rate of positivity for C. trachomatis and the number of endocervical cells (P = 0.24). The rate of positivity for C. trachomatis was 5.4% (8 of 147 specimens) among specimens containing large numbers of erythrocytes (> or =100 per high-power field), whereas it was 6.6% (98 of 1,486 specimens) among specimens containing less than 100 erythrocytes per high-power field (P = 0.59). These results show that the sensitivity of the Abbott C. trachomatis LCR test is affected by the presence of endocervical cells. Additionally, they indicate that the presence of a single endocervical cell is as good an indicator of specimen adequacy as the presence of many endocervical cells. The presence of a large number of erythrocytes was not associated with an increased rate of sensitivity of the LCR.

Cervix Uteri↗

PubMed2003

Detection of Chlamydia trachomatis and Neisseria gonorrhoeae by strand displacement amplification and relevance of the amplification control for use with vaginal swab specimens.

Vaginal swab specimens may be preferable to cervical swab or urine specimens for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae because of the ease of specimen collection and transport. The purpose of this study was to evaluate whether vaginal swab specimens are equivalent to cervical swab specimens for the detection of N. gonorrhoeae and C. trachomatis by the Becton Dickinson strand displacement amplification assay (SDA) with the BDProbeTec ET instrument and then to evaluate the use of the amplification control in a clinical research setting. In the first phase, vaginal and cervical swab specimens were obtained from 455 symptomatic women aged 18 to 40 attending primary health care and sexually transmitted disease clinics. Thirty-nine specimens (8.6%) had true-positive results for N. gonorrhoeae and 37 specimens (8.1%) had true-positive results for C. trachomatis. The sensitivity of SDA was superior to that of culture for the detection of N. gonorrhoeae with vaginal swab specimens and equivalent to that of the Roche PCR for the detection of C. trachomatis with cervical swab specimens. In the second phase of the study, 1,411 consecutively collected vaginal swab specimens were evaluated, with 357 (25.3%) specimens giving indeterminate readings on the basis of the result for the amplification control. The prevalences of sexually transmitted pathogens in vaginal swab specimens with and without use of the amplification control were 6.0 and 5.8%, respectively, for C. trachomatis and 3.1 and 3.0%, respectively, for N. gonorrhoeae. Although, vaginal swab specimens were equivalent to cervical swab specimens for the detection of N. gonorrhoeae and C. trachomatis by SDA with respect to sensitivity, one in four vaginal swab specimens yielded an indeterminate result when the amplification control was used. The amplification control has limited value for use with vaginal swab specimens.

Cervix Uteri↗

PubMed2000

Effects of specimen length on the monosegmental motion behavior of the lumbar spine.

STUDY DESIGN: An in vitro biomechanical analysis of the segmental motion behavior of the same segments in polysegmental (five segments), bisegmental, and monosegmental specimens using sheep lumbosacral spines. OBJECTIVES: To investigate the effect of specimen length on monosegmental motion behavior. These data may be helpful in planning in vitro tests and in comparing results of studies using specimens of different lengths. SUMMARY OF BACKGROUND DATA: The length of spinal specimens used for in vitro stability tests varies greatly, depending on the purpose of the study. Some investigators prefer testing specimens with one adjacent segment on either end of the region of interest. Others favor specimens as short as possible. METHODS: In a first step, seven sheep spine specimens, L3-S1 (note that sheep spines normally have seven lumbar vertebrae), each were tested without preload in a spine-loading apparatus. Alternating sequences of pure lateral bending, flexion/extension, and axial rotation moments (+/-3.75 Nm) were applied continuously. The motion in each single segment was measured simultaneously. Then, these polysegmental specimens were cut into two bisegmental specimens, L3-L5 and L6-S1, and tested in the same way. Finally, another vertebra was removed to obtain two monosegmental specimens, L3-L4 and L7-S1, and to test them as described. RESULTS: In general, the range of motion at L3-L4 and L7-S1 was smaller when tested in polysegmental than in monosegmental specimens. In polysegmental specimens (five segments), the range of motion at L3-L4 and L7-S1 was approximately 80% (range, 70.6-92.5%) and in bisegmental specimens approximately 95% (range, 66.7-100%) of their range of motion measured in monosegmental specimens. Neutral zone and coupled motions showed the inverse behavior. Significant differences were found. However, they were not consistent with either the loading direction or with the specimen length. CONCLUSIONS: For comparison of results, the specimen length should be kept constant within one experiment. Segmental motion behavior of specimens with different lengths should be compared only qualitatively.

Animals↗

PubMed1991

Performance of cryptococcus antigen latex agglutination kits on serum and cerebrospinal fluid specimens of AIDS patients before and after pronase treatment.

Cryptococcal antigen titers in 97 serum and 42 cerebrospinal fluid (CSF) specimens from 37 AIDS patients with culture-proven cryptococcal infection were determined with the Meridian kit (Meridian Diagnostics Inc., Cincinnati, Ohio) before and after treatment with pronase. The geometric mean titers before and after pronase treatment were 1:45 and 1:588 in serum and 1:97 and 1:79 in CSF, respectively. Only on serum (but not CSF) specimens after pronase treatment were (i) titers increased by 2 to 13 dilutions on 57% of the specimens, all of which had titers of less than or equal to 1:128 before pronase treatment, (ii) false-negative reactions on 27% of specimens before pronase treatment eliminated, all of which had titers from 1:4 to 1:4,096, (iii) prozone-like reactions (titer, less than or equal to 1:256) on 9% of the specimens before pronase treatment eliminated, and (iv) agglutination reactions on all specimens stronger and easier to interpret. Antifungal agents added to serum as well as freeze-thaw cycles did not change antigen titers in serum. After two separate tests, the same titers were obtained on 94% of 35 serum specimens that were treated with pronase and on 96% of 53 CSF specimens that were not treated with pronase. A total of 26 serum specimens and 28 CSF specimens from patients with no cryptococcal disease were negative before and after pronase treatment. The IBL kit (International Biological Labs Inc., Cranbury, N.J.) was compared with the Meridian kit on 41 serum specimens and 14 CSF specimens. Results from the two kits agreed on 54 and 68% of serum specimens and 86 and 93% of CSF specimens before and after pronase treatment, respectively. The IBL kit generally produced higher titers on specimens in disagreement and produced no prozone-like reactions. Routine pronase treatment of serum is recommended with the Meridian kit in order to eliminate false-negative and unclear agglutination reactions by producing a consistent interpretation of agglutination reactions. CSF specimens do not require pronase treatment. Titer results produced by the kits from the two different manufacturers varied considerably: the kits should not be used interchangeably for determining antigen titers in serum specimens.

Acquired Immunodeficiency Syndrome↗

PubMed2003

Specimen processing and concentration of Chlamydia trachomatis added can influence false-negative rates in the LCx assay but not in the APTIMA Combo 2 assay when testing for inhibitors.

Inhibitors in clinical specimens can be detected by adding the target of nucleic acid amplification to the sample. Introduction of a Chlamydia trachomatis L2 434 preparation containing 12 elementary bodies (EBs) into first-void urine (FVU) from 225 nonpregnant women and 190 pregnant women before specimen processing by the assays produced false-negative rates of 0.48% (2 of 415 specimens) and 13% (44 of 338 specimens) by the APTIMA Combo 2 and the Chlamydia LCx tests, respectively. Reducing the amount of C. trachomatis added to one EB, a concentration closer to the APTIMA Combo 2 test cutoff, for a subset of 244 FVU specimens increased the number of specimens with false-negative results by the APTIMA Combo 2 assay to 7 (2.9%), suggesting that the strength of the input C. trachomatis per specimen has an influence on the number of specimens with false-negative results. Repeat testing after overnight storage and dilution decreased the APTIMA Combo 2 test false-negative rates to 0% (0 of 415 specimens) with the stronger inoculum and 0.8% (2 of 244 specimens) with the weaker inoculum; the false-negative rate of the LCx assay was reduced to 5.4% (18 of 334 specimens). When an additional 70 FVU specimens from women to which 12 EBs were added before specimen processing were tested by the LCx assay, 34 specimens had false-negative results, whereas 21 specimens had false-negative results when the C. trachomatis EBs were introduced after processing. Nine of the 21 specimens to which EBs were added after processing and all of the 34 urine specimens to which the target was added before processing remained falsely negative on repeat testing at a 1:2 dilution, suggesting that input C. trachomatis DNA was lost during processing by the LCx assay. In contrast, the APTIMA Combo 2 assay appears to have a higher sensitivity and either lost little nucleic acid during processing or demonstrated few problems with inhibitors of transcription-mediated amplification.

Adult↗

PubMed1998

Predictive value of specimen radiography for core needle biopsy of noncalcified breast masses.

OBJECTIVE: Our objective was to determine the predictive value of specimen radiography for large core (14-gauge) needle biopsy of noncalcified breast masses. SUBJECTS AND METHODS: Eighty-four biopsies of 83 breast masses yielded 403 specimens. Specimens showing dense material on specimen radiography were predicted to be diagnostic; specimens showing intermediate- or low-density material were predicted to be nondiagnostic. Specimen radiographic and histopathologic findings were correlated for each specimen using vital dyes to mark individual specimens. RESULTS: Of the 403 specimens, 307 (76%) contained diagnostic material representative of the lesion, with a specific diagnosis achieved for 82 (99%) of 83 lesions (62 benign, 20 malignant). Of the 293 passes containing dense material, 268 (91%) proved to be diagnostic; 11 (18%) of 62 specimens containing only low-density material proved to be diagnostic. Of the 25 (9%) of 293 specimens containing radiographically dense but nondiagnostic material, 18 (72%) showed focal fibrosis and had missed the lesion; 15 (83%) of 18 such specimens were obtained in dense parenchyma. The positive predictive value of specimen radiography was 13 (100%) of 13 in fatty breasts; 77 (96%) of 80 in breasts with minimal scattered fibroglandular elements; 91 (94%) of 97 in heterogeneously dense breasts; and 35 (70%) of 50 in breasts with extremely dense parenchyma. Of the 16 lesions sampled stereotactically, specimen radiography helped assess the inadequacy of initial sampling in three (19%). In six (9%) of 68 sonographically guided biopsies, only one or two specimens could be obtained; specimen radiography helped us predict whether material was adequate for diagnosis. CONCLUSION: Radiography of core specimens obtained from noncalcified breast masses accurately reveals the adequacy of sampling unless the breast parenchyma is extremely dense. Such immediate assessment can help ensure adequate material from lesions that are difficult to biopsy and can thereby improve the diagnostic yield of large core needle breast biopsy.

Adolescent↗

PubMedDate not supplied

Confirmatory testing demonstrates that false-positive rates in the chlamydiazyme assay are influenced by gender and genital specimen type.

BACKGROUND AND OBJECTIVES: Chlamydia trachomatis antigen testing of clinical specimens is replacing culture as the test of choice. Because of a potential for false positive results in low prevalence populations, there is an apparent need for confirming specimens positive by enzyme immunoassay (EIA). GOAL OF THIS STUDY: To examine specimens falsely positive in the Chlamydiazyme EIA assay according to gender and specimen type. STUDY DESIGN: Testing of genitourinary specimens from men and women consecutively enrolled from five health care delivery sources in an urban Canadian population. All specimens were initially tested in the Chlamydiazyme test and all positives repeated in a confirmatory blocking assay provided by the manufacturer. Additional confirmatory testing was performed using immunofluorescence (IF) staining for C. trachomatis elementary bodies (EB's) and polymerase chain reaction (PCR). RESULTS: From Jan. 1, 1990 to June 1, 1991, multiple specimens from 656 men and 5,628 women of varying population prevalences were screened. EIA-positive specimens from women had a repeat negative rate of 22% to 27% from cervical swabs and 29% from urethral swabs. Male urethral swabs had a high repeat negative rate of 22% when EIA was the only positive test, but 2.4% when the specimen was positive by EIA and culture. EIA-positive first void urine (FVU) specimens from men had a repeat negative rate of 8.7% as opposed to 17% to 32% from women. Only 1.7% (2/115) of male FVU did not block compared to rates of 47% (22/47) to 80% (4/5) in FVU from women. Analysis of EIA optical densities (OD's) and EB counts showed an association between the absorbance range 0.1 to 1.4 OD and 0-85 EB's. The greatest number of EB's and highest OD's were seen with cervical specimens, followed by urine and urethral specimens in women infected at all three specimens. All 55 specimens that did not confirm in the blocking test had no EB's and a convenience sample of seven were negative by PCR. All of a subset of 50 blocked specimens contained EB's or were positive by PCR. CONCLUSIONS: Although a variable proportion of specimens may not repeat positive in the EIA, use of the blocking reagent to confirm the repeat positives is highly recommended and the rate of blocking may be heavily influenced by gender and specimen type.

Antigens, Bacterial↗

PubMed1999

Comparison of direct and concentrated acid-fast smears to identify specimens culture positive for Mycobacterium spp.

Microscopic examination of respiratory specimens for acid-fast bacilli (AFB) plays a key role in the initial diagnosis of tuberculosis, monitoring of treatment, and determination of eligibility for release from isolation. The objective of this study was to compare the sensitivity obtained with smears for detection of AFB (AFB smears) made directly from respiratory specimens (direct AFB smears) to that obtained with parallel smears made from concentrates of the specimens (concentrated AFB smears). A total of 2,693 specimens were evaluated; 1,806 were from the University of California Irvine Medical Center Medical Microbiology Laboratory (UCIMC), which serves a tertiary-care hospital with outpatient clinics, and 887 were from the Microbial Disease Laboratory at the California Department of Public Health (MDL), which receives specimens from outpatient facilities and clinics on Pacific islands. Of the 353 AFB culture-positive specimens at UCIMC, there was a statistically significant difference in the sensitivity of the direct AFB smear (34%) and that of the smear made from the concentrated specimen (58%) (P < 0.05). This was also true for the 208 specimens positive for Mycobacterium tuberculosis, for which the sensitivity of the direct smear was 42% (87 of 208) and that for the smear made from the concentrated specimen was 74% (154 of 208). At MDL, where all but 1 of the 45 culture-positive specimens grew M. tuberculosis, the sensitivity of the smear made from the concentrated specimen was 93% (42 of 45) and was not significantly higher than the sensitivity of the direct smear, which was 82% (37 of 45). By combining the results from both laboratories, 42 patients from whom at least three specimens were received were culture positive for M. tuberculosis. The cumulative results for the initial three specimens from these patients showed that the direct smear detected M. tuberculosis in 81% of these patients, whereas the smear made from the concentrate detected M. tuberculosis in 91% of these patients. In summary, when all culture-positive specimens are considered, the sensitivity of the direct smear compared to that of a smear made from the concentrated specimen was significantly different overall in the two different laboratory settings. However, this difference was reduced only if the cumulative results for the initial three specimens received from patients who were culture positive for M. tuberculosis were evaluated.

Bacteriological Techniques↗

PubMed2003

Diagnostic quality of percutaneous kidney biopsy specimens obtained with laparoscopy versus ultrasound guidance in dogs.

OBJECTIVE: To compare diagnostic quality of percutaneous kidney biopsy specimens obtained with laparoscopy versus ultrasound guidance in dogs and compare diagnostic quality of specimens obtained with 14- versus 18-gauge biopsy needles. DESIGN: Prospective study. ANIMALS: 10 healthy dogs. PROCEDURE: In each dog, 2 biopsy specimens were obtained from each kidney, 1 with a 14-gauge biopsy needle and 1 with an 18-gauge biopsy needle. Biopsy specimens were obtained from 1 kidney by means of ultrasound guidance and from the contralateral kidney by means of direct viewing during laparoscopy. Number of glomeruli, quality of the biopsy specimen, proportion of specimens that contained muscle tissue, and proportion of specimens with fragmentation or crushing were determined. RESULTS: Mean +/- SD number of glomeruli (32.6 +/- 11.0) in laparoscopic, 14-gauge biopsy specimens was significantly higher than mean number of glomeruli in ultrasound-guided, 14-gauge specimens; mean number of glomeruli in ultrasound-guided, 18-gauge specimens; and mean number of glomeruli in laparoscopic, 18-gauge specimens. All 10 laparoscopic, 14-gauge biopsy specimens were classified as excellent. The proportion of 18-gauge biopsy specimens with crushing or fragmentation was significantly higher than the proportion of 14-gauge specimens. One of the kidneys biopsied with ultrasound guidance had a large amount of hemorrhage. Hemorrhage was modest and transient following laparoscopic biopsy. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that excellent-quality renal biopsy specimens with large numbers of glomeruli can be obtained with 14-gauge, double-spring-activated biopsy needles during laparoscopy. Renal biopsy specimens obtained with 18-gauge biopsy needles frequently had few glomeruli and often were crushed or fragmented, increasing the difficulty in making an accurate diagnosis.

Animals↗

PubMed1996

Effect of specimen fixation method on pullout tests of pedicle screws.

STUDY DESIGN: Experimental axial pullout tests of a new type of pedicle screw were done on cadaveric lumbar vertebrae. The manner in which specimens were secured in the testing apparatus was varied to determined influence of specimen fixation method on the maximum pedicle screw pullout force. OBJECTIVES: To determine the appropriateness of embedding (i.e., potting) spinal specimens in polymer resin (e.g., bone cement or Plastic Padding [Plastic Padding Ltd., High Wycombe, Buckinghamshire, England]) for axial pullout tests of pedicle screws. Several different specimen fixation methods were examined to make recommendations for the standardization of future experimental testing protocols. SUMMARY OF BACKGROUND DATA: Axial pullout of transpedicular screws, although not a likely clinical mode of failure, is a popular experimental testing mode for evaluating screw-bone biomechanics. A wide variety of techniques for securing a vertebral specimen to counter the axial pullout force has been reported (including the use of polymer resin) with a correspondingly wide range in the resulting axial pullout strengths. The possible influence of the specimen fixation method on pedicle screw axial pullout strength has not been addressed previously. METHODS: Axial pullout tests of pedicle screws (DDS, Plus Endoprothetik, Rotkreuz, Switzerland) from the pedicles of 21 isolated lumber vertebral bodies were done using a Model 810 MTS Universal Testing Machine (MTS Systems, Inc., Minneapolis, Minnesota). The specimens were secured in a custom-made vise fixture either as is or after the vertebral bodies were potted in Plastic Padding up to the pedicle origin. Some of the potted specimens were wrapped first in latex to prevent polymer resin intrusion, and the others were unprotected. Pullout tests were attempted on both the left and right pedicles of each specimen, and the maximum pedicle screw pullout force was recorded. Measurement of bone mineral density by means of dual energy x-ray absorptiometry, in addition to macroscopic and scanning electron microscopy histologic analyses, microradiography, and energy dispersive X-ray spectroscopy, was done post-test to assist in the interpretation of the data. RESULTS: The maximum pedicle screw pullout force was found to be dependent on both the bone mineral density and the mode of fixation of the vertebrae. Embedding in polymer resin without protection of the specimen (i.e., latex wrapping) led to several instances of well-documented polymer resin intrusion; in these specimens, mean maximum pedicle screw pullout force was significantly greater than that of specimens secured without polymer resin and that of embedded specimens for which intrusion did not occur. CONCLUSIONS: Polymer resin intrusion can have a significant effect on the biomechanical characteristics of the bone-pedicle screw interface. When polymer resins are used to secure vertebral specimens for in vitro biomechanical tests of the bone-pedicle screw interface, it is important to either prevent intrusion (e.g., with a latex wrapping) or document post-test (e.g., through the methods described in this article) that intrusion did not occur for the specimens included in the analysis.

Aged↗

PubMed1984

Results of analyses and bacterial cultures of urine specimens obtained from clinically normal cats by three methods.

Six urine specimens were obtained from each of 6 male and 6 female cats in a 3-week period. The first and last specimens from each cat were obtained by cystocentesis, 2 were obtained by urethral catheterization, and 2 were caught during voiding stimulated by manual bladder compression. Quantitative urine cultures did not reveal bacteriuria in specimens obtained by cystocentesis, and urinary tract infection did not develop during the study. Bacteria were found in 3 (25%) of 12 urine specimens obtained by catheterization of males and in 1 (8%) of 12 specimens obtained by catheterization of females. Magnitude of bacteriuria in specimens obtained by catheterization was 10 to 1,000 organisms/ml. Bacteria also were found in specimens obtained during voiding. Each of 11 (100%) cleanly caught specimens obtained from males and 7 (58%) of 12 voided specimens from females contained bacteria. Magnitude of bacteriuria in voided specimens was usually 100 to 10,000 organisms/ml, but 3 voided specimens contained greater than 10,000 organisms/ml. Many bacteriuric specimens contained more than 1 type of organism; however, bacteria that might be suspected of causing urinary tract infections in cats were found frequently. Urinalyses were performed on 66 specimens. Completeness of urinalyses depended on the volume of specimens available, but results were normal except for evidence of hematuria in a few specimens obtained by cystocentesis or catheterization. Hematuria was usually mild and was attributed to hemorrhage caused by minor urinary tract trauma during urine collection.

Animals↗

PubMed2005

Clear-cell adenocarcinoma of the female genital tract: presence of hyaline stroma and tigroid background in various types of cytologic specimens.

Hyaline basement membrane-like stromal material and tigroid background are distinctive cytologic features observed in Diff-Quik (DQ)- or Giemsa-stained smears of clear-cell adenocarcinoma (CCA) of the female genital tract. However, it is uncertain how often these features are present in different types of cytologic specimens, and which type of preparation is optimal for this diagnosis. We therefore reviewed the cytologic features of CCA in three types of specimens, including 15 scrape cytology specimens, 7 fine-needle aspiration (FNA) specimens, and 15 peritoneal cytology specimens, with emphasis on the features observed in DQ-stained smears. The cell morphology in scrape cytology specimens and FNA specimens was comparable, whereas in peritoneal cytology specimens, the cytoplasm was better preserved. Most tumor cells had fragile cytoplasm containing variable amounts of fine vacuoles, and round nuclei with distinct or prominent nucleoli. Hyaline stroma was present in 93% of scrape cytology specimens, 71% of FNA specimens, and 80% of peritoneal cytology specimens. Tigroid background was observed in 47% of scrape cytology specimens, 43% of FNA specimens, but in none of the peritoneal cytology specimens. Formation of a tigroid background may be prevented by the abundant fluid content in peritoneal cytology specimens. Hyaline stroma and tigroid background were uncommonly seen in scrape smears from other types of primary ovarian tumors, mainly juvenile granulosa cell tumor and yolk sac tumor. However, the additional presence of papillary structures allows CCA to be readily distinguished from these other tumors. We propose that scrape cytology offers the best approach for the intraoperative cytologic diagnosis of CCA.

Adenocarcinoma, Clear Cell↗

PubMed1999

Stability study on specimens mailed to a state laboratory and tested with the Gen-Probe PACE 2 assay for chlamydia.

BACKGROUND: Although specimen collection is acknowledged to be a critical factor in the testing of chlamydia, rarely do studies examine the effects of specimen transport on laboratory results. GOAL: To compare the results on specimens shipped in a controlled environment with duplicate specimens exposed to environmental conditions such as heat or extended time in transit. STUDY DESIGN: Duplicate specimens were collected from 1,017 women tested at South Carolina public health clinics. One specimen from each woman was shipped by courier and the other by U.S. mail. The results, swab collected first, method of transport, and temperature during shipment were compared for each set of specimens. Specimens were tested with the Gen-Probe PACE 2 test. RESULTS AND CONCLUSIONS: Mailed specimens were exposed to longer transport times and elevated temperatures; despite this, 99% of the results using courier specimens agreed with the mailed specimen results. Eighty-eight women tested positive and 891 women tested negative for chlamydia on both specimens. When the 11 specimens with discrepant results were retested by polymerase chain reaction, 10 were positive for chlamydia, with 9 concurring with the mailed specimen results. Results of specimens in this study were not adversely affected by heat and extended transit times when transported by U.S. Mail.

Adult↗