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PubMed1991

Expression of leucocyte adhesion molecules on 66 clinical neuroblastoma specimens.

LFA-3, ICAM-1, HLA.ABC and HLA.DR expression was analyzed on 66 neuroblastoma specimens. HLA.ABC was expressed on 26 specimens, HLA.DR on 2, LFA-3 on 20 and ICAM-1 on 10. HLA.ABC and LFA-3 were positive on ganglioneuroblastoma or ganglioneuroma, but they were negative on neuroblastoma, independently of the clinical staging; HLA.ABC and LFA-3 were induced in vivo by chemotherapy in parallel with tumoral cell differentiation, in both the primary and the metastases. The expression of ICAM-1 was restricted to 5 of the 10 low-grade stage-1 or stage-2 specimens, 1 stage-3 specimen, and the primary tumors of 2 patients with stage-4 disease, analyzed hence at diagnosis and after chemotherapy (4 specimens); metastatic cells obtained in 1 of these patients were negative. HLA.ABC and LFA-3 expressed on both mycN-negative and -positive specimens, whereas ICAM-1 was restricted to MYCN-negative specimens. LFA-3 diffusely stained partially differentiated neuroblasts, Schwann cells and ganglion cells. The expression of HLA.ABC on differentiated neuroblasts varied from one sample to another and within the same tumor; Schwann cells were strongly positive, but ganglion cells were negative. In positive samples, ICAM-1 was expressed on differentiated neuroblasts and Schwann cells, but negative on ganglion cells; however, most of the differentiated tumors were ICAM-1-negative, suggesting ICAM-1 induction by unknown local signal. The 4 markers were negative on undifferentiated neuroblasts. The distribution of these 4 markers on clinical specimens was in agreement with their reactivity on fetal tissues, as well as with results obtained on neuroblastoma cell lines before and after in vitro treatment with IFN-gamma.

Alkaline Phosphatase↗
PubMedDate not supplied

The influence of temperature and specimen size on the flexural properties of PMMA bone cement.

A three-stage investigation was undertaken to examine the influence of specimen size and polymerization environment on the flexural properties of PMMA. In the first stage, specimens were cut from large pieces of commercially cast acrylic (lucite) and self-curing dental acrylic. The material properties of the cast acrylic as calculated from the results of flexural testing were not influenced by specimen size in the range of 1 X 10 X 30 to 3 X 10 X 60 mm, whereas 1-mm thick dental cement specimens had 12% (p less than 0.001) higher stress and 24% (p less than 0.001) higher strain to failure than 3-mm specimens cut from the same sample. In the second stage, self-curing dental cement and Simplex P bone cement were molded in different thicknesses. These experiments demonstrated that molding thin 1.6-mm specimens resulted in increases of 14% (p less than 0.001) in stress and 30% (p less than 0.001) in strain to failure as compared with 3.2-mm specimens, which were greater differences than those due to the specimen thickness effects alone. In the third stage, temperatures of cement were monitored during polymerization, and cements were molded at different temperatures. These results demonstrated that thicker cement samples reached higher temperatures and that cement samples polymerized at lower temperatures (21 versus 37 degrees C) had 12% (p less than 0.001) greater stress and 105% (p less than 0.001) greater strain to failure.

Bone Cements↗
PubMed1981

Detection of influenza A virus by radioimmunoassay and enzyme-immunoassay from nasopharyngeal specimens.

Four-layer (indirect) radioimmunoassay (RIA) and enzyme-immunoassay (EIA) techniques were developed for the detection of influenza A and B virus in the sonicated nasopharyngeal specimens from patients hospitalized for acute respiratory infection. Polystyrene beads (RIA) or polystyrene microtiter plates (EIA) were used as the solid-phase, guinea pig antivirus immunoglobulins as the catching antibodies, rabbit antivirus immunoglobulins as the secondary antibodies, and 125I-labeled sheep antirabbit (RIA) or horseradish peroxidase conjugated swine antirabbit (EIA) immunoglobulins as the detector antibodies. A comparison of the developed RIAs and EIAs with the immunofluorescence (IF) method was made with 41 influenza A IF-positive and 150 influenza A IF-negative specimens. Each of the 41 influenza A IF-positive specimens was positive by the influenza A RIA and negative by the influenza B RIA. Out of 150 influenza A IF-negative specimens 3 specimens were found with weakly positive results in influenza A and B RIAs, but in each of these 3 specimens the binding proved nonspecific by the corresponding confirmatory tests. Using the EIa technique and the same immunoreagents as in RIA, identical results were obtained in each selected specimen tested. The developed RIAs and EIAs proved to be as specific and sensitive as the IF technique, and they should be practical in the diagnosis of respiratory infections directly from nasopharyngeal specimens.

Adolescent↗
PubMed1990

Uterine cervical punch biopsy specimens can be analyzed by 1H MRS.

Biopsy specimens of the uterine cervix, including colposcopically directed punch biopsy specimens of females with atypical Papanicolaou smear tests, are suitable for analysis by magnetic resonance (MR) spectroscopy. A narrow lined lipid MR spectrum, characteristic of malignant tissue, is obtained from a 6-mm3 biopsy specimen of histologically confirmed squamous carcinoma of the cervix. In contrast, specimens containing inflammatory cells generate a broad component only centered at 1.3 ppm with a T2 relaxation value of less than 350 ms. Most biopsy specimens which contain dysplastic cells or evidence of human papilloma virus (HPV) infection have a discernible lipid spectrum similar to that of the malignant tissue specimen. Long T2 relaxation values found in malignant tissue specimens at 1.3 and 1.2 ppm are observed in some but not all of the biopsies which show evidence of HPV infection. The suitability of small tissue samples, such as punch biopsy specimens, for study by MR illustrates the sensitivity of this technique and its potential as an aid to histopathological discrimination between the various precursor states of cervical cancer.

Biopsy↗
PubMed1999

Direct detection of vanA and vanB genes in clinical specimens for rapid identification of vancomycin resistant enterococci (VRE) using multiplex PCR.

Surveillance for vancomycin resistant enterococci (VRE) by culture can be labour intensive and time consuming. We have developed a multiplex polymerase chain reaction (MPCR) which can be performed directly on the clinical specimen. The assay allows sensitive detection of enterococci with vanA - and vanB -mediated resistance to vancomycin. DNA was purified from stool and rectal specimens using the XTRAX(TM)DNA Extraction Kit (Gull Labs). Multiplex PCR amplified vanA and vanB targets were detected using a microtiter plate EIA. Two-hundred specimens were tested by routine culture and MPCR. Culture identified 44 VRE isolates and MPCR detected 38 of the 44 culture positives. Multiplex PCR detected three additional positive VRE specimens missed by culture for a sensitivity and specificity of 86.4 and 98.1%, respectively. When the presence of PCR inhibitors was addressed in the six culture positive/MPCR negative specimens, four additional VRE positive specimens were detected. Performing MPCR on the original specimens and on a 1:10 dilution of all specimens to minimize the effect of inhibitors gave a sensitivity and specificity of 95.5 and 98.1%, respectively. Multiplex PCR with confirmation by microtiter plate hybridization could be completed in 8 h compared with 24-48 h required for culture.

Bacterial Proteins↗
PubMed1990

Spatial and temporal continuity of kangaroo rat populations shown by sequencing mitochondrial DNA from museum specimens.

The advent of direct sequencing via the polymerase chain reaction (PCR) has opened up the possibility of molecular studies on museum specimens. Here we analyze genetic variation in populations over time by applying PCR to DNA extracted from museum specimens sampled from populations of one species over the last 78 years. Included in this study were 43 museum specimens of the Panamint kangaroo rat Dipodomys panamintinus from localities representing each of three geographically distinct subspecies. These specimens were originally collected and prepared as dried skins in 1911, 1917, or 1937. For each specimen, a 225-bp segment of the mitochondrial genome was sequenced. These mitochondrial DNA sequences were compared to those of 63 specimens collected at the same localities in 1988. The three subspecies were nearly completely distinct. Only 2 of the 106 individuals shared mitochondrial types between subspecies. For all three localities, the diversity levels were maintained between the two temporal samples. The concordance observed between the two temporally separate phylogenies supports the use of museum specimens for phylogenetic inference. This study demonstrates the accuracy and routine nature of the use of museum specimens in the analysis of mitochondrial sequence variation in natural populations and, importantly, that a temporal aspect can now be added to such studies.

Animals↗
PubMed2004

Ultrasound during dissection of cadaveric specimens: a new method for obtaining ultrasound-anatomic correlations in musculoskeletal radiology.

We report a new technique for ultrasound-anatomic correlations consisting of dissection of embalmed specimens during ultrasound examination. Our method consists of performing ultrasound during the different stages of dissection. The technique was developed by making observations of selected structures in two embalmed and two non-embalmed cadaver hands. The image quality was subjectively graded by consensus of two investigators, before and after denudation of the selected structures of the hand. As an example, the technique is demonstrated for the flexors at the metacarpophalangeal joint level, the extensor complex at the level of the proximal phalanx, and the dorsal hood of the second to fourth fingers. Before dissection the image quality in fresh specimens was graded moderate, and in embalmed specimens good. After dissection the image quality was good in fresh specimens and excellent in embalmed specimens. Our method is simple and does not require sophisticated material. Our results indicate that embalmed specimens could be better than non-embalmed specimens, because of the presence of artefacts in the non-embalmed specimens (gas deposits). The described methodology can yield excellent results regarding precise identification of different interfaces and structures, as observed at ultrasound.

Cadaver↗
PubMed1999

Extracellular matrix-induced cell migration from glioblastoma biopsy specimens in vitro.

The present knowledge about the interaction between the extracellular matrix (ECM) and gliomas is mostly based on studies of permanent cell lines. Since such cultures have undergone an extensive clonal selection in vitro, the experimental results obtained may be quite different from those obtained from studies on true biopsy specimens. The present work demonstrates how different ECM components affect tumor cell migration from human glioblastoma specimens grown as biopsy sample spheroids. Biopsy specimens from 12 glioblastomas and 1 gemistocytic astrocytoma were included in this study. Spheroids were directly initiated from the biopsy specimens, and after 3-4 weeks in culture, they were used in a migration assay. A custom-made filtered medium, where the high molecular weight (>100 kDa) proteins were removed, was supplemented with the following ECM components: laminin, fibronectin, collagen type IV and vitronectin. The cell migration was negligible when spheroids were propagated in the filtered medium. The ECM components as well as complete DMEM evoked strong stimulatory effects on different biopsy specimens. Opposed to that observed earlier for permanent glioma cell lines, highly variable responses were observed between the different biopsy samples on the various ECM components. In general, correlation analyses revealed that specimens that were strongly stimulated by laminin were also stimulated strongly by fibronectin, collagen type IV and vitronectin. This suggests that the capacity to migrate as a response to ECM was confined more to each biopsy specimen than to any specific ECM component. Since biopsy sample spheroids, as original tumors, consist of different cell types, an immunohistochemical characterization of the migrating cells was also performed. Anti-glial fibrillary acidic protein (GFAP) staining revealed both GFAP-positive and -negative migrating cells. Immunostaining for von Willebrand factor and CD11b indicated that the migrating cells were neither endothelial nor microglial cells. This study, therefore, indicates that migratory responses of glioma biopsy specimens to different ECM components is much more heterogeneous than that observed earlier for cell lines. Furthermore, the presented findings support the notion that gliomas may utilize different cell surface receptors for their migration, depending on the cell substrates available.

Adult↗
PubMed2003

Feasibility of histological grading and staging of chronic viral hepatitis using specimens obtained by thin-needle biopsy.

We performed a retrospective study to investigate the feasibility of grading and staging of chronic viral hepatitis on specimens obtained by means of thin-needle biopsy (TNB; 20G) using the modified Ishak-system (J Hepatol 1995; 22:696-699). Specimens obtained using large-needle biopsy (LNB; 17G) served as a control. A total of 100 biopsy specimens from 88 patients were included in the study. Of the patients, 30 suffered from chronic hepatitis B, 54 from chronic hepatitis C, and 4 from both; 59 specimens were obtained by TNB and 41 by LNB. All four categories of the Ishak-system, i.e., interface hepatitis, confluent necrosis, lobular inflammation and portal inflammation, could be applied to TNB specimens and provided similar total scores to those observed in LNB specimens. Specimens obtained by TNB facilitated the diagnosis of liver cirrhosis. However, they bore the risk of underestimating the presence of cirrhosis in favor of advanced bridging fibrosis, whereas no differences were found in the overall recognition of liver fibrosis. Intra- and interobserver variabilities were not affected by the needle size. For the interobserver agreement, the kappa values for the category of inflammation ranged from 0.003 to 0.419 (TNB) and 0.096 to 0.470 (LNB) and for staging we noted kappa values of 0.351 (TNB) and 0.456 (LNB). Reproducibility increased when a tolerance of +/-1 was accepted for grading (total score) and staging; in this case, observer variability was less than 20%. This study showed that grading and staging of chronic viral hepatitis is feasible in TNB specimens and that intra- and interobserver variability poses a greater problem than needle size.

Adult↗
PubMed2005

Renal transplant biopsy specimen adequacy in a paediatric population.

Updated guidelines on the diagnosis of acute allograft rejection including criteria for biopsy specimen adequacy were published in 1999. We sought to determine the adequacy of specimens in paediatric transplant patients and identify factors influencing adequacy. All renal transplant biopsies performed between 1998 and 2003 were classified as adequate (n =25), minimal (n =19) or inadequate (n =27) in accordance with the Banff 97 criteria, and the histological diagnoses were documented. The effect on specimen adequacy of grade of operator, method of sedation, age of child, needle gauge, number of cores and total core length was then investigated. Overall, a minimal or adequate specimen was obtained in 62% of cases. No histological diagnosis could be made in 30% of all specimens, just over half of which were inadequate. Higher rates of rejection were found in adequate (52%) than inadequate (33%) samples. The grade of operator (p =0.498), the age of the child at the time of biopsy (p =0.815) and type of sedation (p =0.188) did not affect adequacy. More than one core was obtained in 38 (54%) cases, and this was significantly associated with specimen adequacy (p <0.0005) as was longer total core length (p =0.002). Clinical features in isolation are not sufficient for the diagnosis of acute allograft rejection. Renal biopsy remains the gold standard and relies on adequate specimen collection. Our data shows that specimen adequacy according to the Banff 97 guidelines is achievable in children and that more than one core at the time of sampling significantly improves this achievement. Adequate sampling reduces the risk of an inconclusive histological diagnosis.

Adolescent↗
PubMed1998

Comparison of the ligase chain reaction with solid and liquid culture media for routine detection of Mycobacterium tuberculosis in nonrespiratory specimens.

The aim of this study was to compare the results of a commercial assay based on the ligase chain reaction [(LCR) LCx Probe System MTB; Abbott, USA] with those of culture in liquid medium (Septi-Chek AFB; Becton-Dickinson, USA) and culture on the egg-based Löwenstein-Jensen solid medium for the direct detection of Mycobacterium tuberculosis complex in nonrespiratory specimens. The results were analyzed according to the standard definition of a true-positive result. Two hundred thirty-five nonrespiratory samples routinely submitted to rule out tuberculosis were analyzed. All samples were smear-negative. Mycobacterial growth in either culture medium was detected in 18 (7.6%) specimens: Mycobacterium tuberculosis was recovered from seven (38.9%) specimens cultured on Löwenstein-Jensen medium and from 18 (100%) specimens cultured in Septi-Chek AFB. The LCR protocol was positive in 22 specimens. None of the LCR-negative controls showed positive results. All samples positive by culture on Löwenstein-Jensen medium were positive by culture in liquid medium and by the LCR assay. However, Mycobacterium tuberculosis was detected by culture in liquid medium in two specimens that were negative by the LCR assay, whereas six specimens negative by culture in liquid medium were positive by the LCR protocol; three of these were identified as true-positive results of the LCR assay. The sensitivity, specificity, and positive and negative predictive values were 33.3%, 100%, 100%, and 93.8%, respectively, for Löwenstein-Jensen medium; 85.7%, 100%, 100%, and 98.6% for the liquid medium; and 90.4%, 98.5%, 86.3%, and 99% for the LCR assay. These findings indicate that the LCR assay may be a valid method of high diagnostic yield for direct detection of Mycobacterium tuberculosis complex in nonrespiratory specimens.

Body Fluids↗
PubMed2002

Predicting 5-FU sensitivity using human colorectal cancer specimens: comparison of tumor dihydropyrimidine dehydrogenase and orotate phosphoribosyl transferase activities with in vitro chemosensitivity to 5-FU.

BACKGROUND: In tumor cells, the enzyme orotate phosphoribosyl transferase (OPRT) contributes to 5-fluorouracil (5-FU) phosphorylation and another enzyme, dihydropyrimidine dehydrogenase (DPD), is associated with 5-FU catabolic action. We measured OPRT and DPD activities and, to determine whether their levels might serve as indicators of 5-FU sensitivity, simultaneously assayed in vitro chemosensitivity to 5-FU. METHODS: Tissue specimens were obtained from colorectal cancer patients and in vitro chemosensitivity was tested using fluorescein diacetate assay (FDA) or histoculture drug response assay (HDRA). DPD and OPRT activities were measured by radioassay. RESULTS: The chemosensitivity assay was performed on 62 colorectal cancer specimens. Results were evaluable in 29 of 30 cases (96.7%) for FDA and 30 of 32 cases (98.3%) for HDRA. The positive sensitivity rate was 37.9% by FDA assay and 30% by HDRA assay. In positive specimens, the mean DPD activity was 44.9 +/- 32.6 pmol/min per mg protein, and in negative specimens, it was 53.8 +/- 33.7 pmol/min per mg protein ( P= 0.875). In contrast, the mean OPRT value was significantly higher in positive specimens (0.418 +/- 0.180 nmol/min per mg protein) than in negative specimens (0.325 +/- 0.153 nmol/min per mg protein; P < 0.05). The chemosensitivity test proved positive in 60% of the specimens with ORPT activity of 0.413 or above and 50% of those with DPD activity of 30 or below. Of the patient specimens showing OPRT activity of 0.413 or above and DPD activity of 30 or below, 88.9% were positive for 5-FU sensitivity, suggesting the possibility that the combination of these two levels may be predictive of 5-FU positive sensitivity. CONCLUSION: DPD and OPRT activities within cancer cells may predict positive sensitivity to 5-FU.

Adult↗
PubMed2000

Role of specimen radiography in patients treated with skin-sparing mastectomy for ductal carcinoma in situ of the breast.

BACKGROUND: Specimen radiography is an important part of breast conservation surgery for ductal carcinoma in situ (DCIS). The objective of this study was to determine whether mastectomy specimen radiography could help in obtaining negative resection margins in patients with DCIS undergoing skin-sparing mastectomy (SSM) with immediate breast reconstruction (IBR). METHODS: Of 95 patients treated at our institution with SSM and IBR for DCIS, 35 had specimen radiography. The mastectomy specimen was first examined grossly and then inked, serially sectioned, and sent for radiographic assessment. Tissue slices containing calcifications were identified for pathologic evaluation. Additional tissue was excised if tumor was found near the inked margins or if calcifications were found near the radiographic margins. RESULTS: Of the 35 patients who had specimen radiography, the radiographic margins were free of calcifications in 30 patients (86%); of these patients, the margins on the final histologic examination were free of tumor in 27 and within 1 mm in 3. The other five patients (14%) had calcifications close to the radiographic margin; four underwent an intraoperative re-excision, but the margin for one of those four patients was still positive on final histologic examination. Margins were found to be negative by both mastectomy specimen radiography and histology in 77% of the patients. Of the 95 patients with DCIS, three patients (3%), none of whom had specimen radiography, developed local recurrences. One of these was successfully re-treated, one died as a result of synchronous distant metastases, and one was lost to follow-up. At a median follow-up time of 3.7 years, 93 patients (98%) were alive and free of disease. CONCLUSIONS: Intraoperative radiography of mastectomy specimens may be useful for assessing margin status and for identifying the location of microcalcifications within tissue slices.

Adult↗
PubMed1990

Evaluation of an enzyme immunoassay for the detection of herpes simplex virus (HSV) antigen from clinical specimens in viral transport media.

A total of 301 clinical specimens (229 culture positive and 72 culture negative) were assayed retrospectively by an enzyme immunoassay. Specimens were transported to the virus lab in viral transport media (VTM), and were inoculated into HF and A549 cell culture tubes for viral isolation with the remainder of the sample being saved at -70 degrees C. Specimens were thawed, vortexed and resuspended in 10 x Herptran concentrate and each sample was then added in duplicate to designated wells of a microtiter plate for the EIA assay. The EIA detected 147/150 (98%) culture positive specimens from symptomatic patients, 63/79 (79.7%) culture positive specimens from patients considered asymptomatic and 210/229 (91.7%) culture positive specimens overall. The EIA was negative for 70/72 (97.2%) culture negative specimens. These data suggest that the EIA test can be used with clinical specimens submitted in conventional VTM. However, VTM samples which are EIA negative, particularly with EIA values close to the EIA positive cutoff value, need to be cultured.

Antigens, Viral↗
PubMed1992

Verification of a formula for determination of preexcision surgical margins from fixed-tissue melanoma specimens.

BACKGROUND: Recently our group reported on the shrinkage of 199 malignant melanoma surgical-excision specimens. In that report, a multivariate analysis revealed that the age of the patient was the only factor that significantly affected the percentage shrinkage of a surgical specimen. In addition, a formula was presented that extrapolates the actual surgical margins (in vivo) from the (contracted) fixed-tissue pathology report measurement and the reported in vivo lesion diameter. OBJECTIVE: The goals of this study are to verify that shrinkage of surgical specimens is approximately 20% and that the margin formula can be successfully applied to a different group of patients. METHODS: Four hundred seven patients with malignant melanoma were prospectively enrolled to measure preexcision (outlined with ink) surgical margins, fixed-tissue (contracted) surgical margins, and overall specimen shrinkage. RESULTS: It is verified that overall shrinkage of cutaneous surgical specimens is approximately 20%. Surgical specimens from patients younger than 50 years of age have approximately 25% shrinkage. Those specimens from patients 50 to 59 years of age have approximately 20% shrinkage and those from patients 60 years of age or older have about 15% shrinkage. The surgical margins predicted by the margin formula were within +/- 3.5 mm of the actual measured surgical margin 86.5% of the time. CONCLUSION: The actual surgical margins (in vivo) of a malignant melanoma can be reasonably estimated from the fixed-tissue pathology measurement via the margin formula. The shrinkage of a surgical specimen is 15% to 25% depending on the patient's age.

Adolescent↗
PubMed1988

Comparison of cell culture and three enzyme-linked immunosorbent assays for the rapid diagnosis of respiratory syncytial virus from nasopharyngeal aspirate and tracheal secretion specimens.

A total of 211 specimens, including 144 nasopharyngeal aspirates and 67 tracheal secretions, were evaluated for the rapid detection of Respiratory Syncytial Virus (RSV) antigen by three commercial enzyme-linked immunosorbent assays (ELISA; Kallestad Diagnostic, Austin, TX; Ortho Diagnostics, Raritan, NJ, and Abbott Laboratories, North Chicago, IL) and by isolation of RSV in cell culture. Of the 61 RSV culture positive specimens, Kallestad ELISA, Ortho ELISA, and Abbott ELISA detected RSV antigen in 80%, 95%, and 92% of the specimens, respectively. An additional 28 specimens were found to be positive only by one or more RSV ELISA tests. A blocking assay confirmed the specificity of ELISA in 71% (20/28) of the RSV ELISA positive and the culture-negative specimens, and 29% were found to be false positive. Through the use of cell culture, with the resolution of ELISA positive and culture negative specimens by blocking assay, 81 specimens (61 + 20) were considered to be true positive. The sensitivity, specificity, and diagnostic accuracy were, for cell culture, 75%, 100%, and 91%; for Kallestad ELISA, 79%, 98%, and 91%; for Ortho ELISA, 95%, 99%, and 98%; and for Abbott ELISA, 93%, 96%, and 95% respectively. In our study, commercial ELISA tests have been shown to be rapid, reliable tests for the detection of RSV. Ortho ELISA and Abbott ELISA showed better sensitivity than the Kallestad ELISA for RSV detection directly in the clinical specimens.

Adolescent↗
PubMed1993

Impact of intervention on the frequency with which leaky urine specimens are received in the laboratory.

Prior to intervention, 4.5% of urine specimens received for culturing in the University of Massachusetts Medical Center clinical microbiology laboratory were noted to have leaked during transport. After an extensive educational program was conducted aimed at alerting physician, nursing, and clerical personnel about the problem, a policy was adopted in which physicians were notified by telephone when the laboratory received leaky specimens. The specimens, however, were processed. Leakage rates dropped to 1.7%. A second round of intervention in which leaky urine specimens were not processed was then implemented. No additional decrease in the rate of receipt of leaky urine specimen was noted (that is, 1.9%). One year later, having adopted a policy of not processing leaky urine specimens, the rate of leaky specimens remained at 1.9%. The clinical microbiology laboratory can effectively intervene regarding problems of specimens that have leaked during transport to the laboratory.

Humans↗
PubMedDate not supplied

Improved detection of mycobacterial antigens in clinical specimens by combined enzyme-linked immunosorbent assay.

Three types of antibodies against cellular and secretory-excretory protein antigens were simultaneously used for the direct detection of mycobacterial antigens in sputum and cerebrospinal fluid (CSF) specimens, using enzyme-linked immunosorbent assay (ELISA). The antibodies consisted of in-house raised and prepared anti-whole-cell, heat-killed, and sonicated Mycobacterium tuberculosis, anti-secretory-excretory protein extract of bacilli Calmette-Guerin (BCG) strain, and commercially available anti-BCG. Sputum specimens comprised 24 smear positive, culture positive, and 47 smear-negative, culture positive (SNCP), from patients with pulmonary tuberculosis, as well as 45 smear-negative, culture-negative (SNCN) control samples. The CSF specimens included 18 SNCPs from patients with tuberculous meningitis and 18 SNCN controls. The sensitivity of the individual tests for sputum and CSF specimens ranged from 70% to 79% and 72% to 89%, respectively, whereas in the combined tests it reached 86%-96% for sputum specimens and 100% for CSF specimens. The specificity of ELISAs for sputum specimens was lower in the combined (73%-87%) than in the individual (87%-98%) tests, whereas for CSF specimens it was 100% in all tests. Thus, the combined ELISA approach for mycobacterial antigen detection provides a rapid and reliable laboratory adjunct in the diagnosis of patients with tuberculosis.

Antibodies, Bacterial↗