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PubMed2002

Appropriate in vitro test conditions for genotoxicity testing of fibers.

With the exception of asbestos fibers, little information is available on genotoxicity testing of fibers (i.e., respirable-sized, fiber-shaped particulates, RFP). In contrast to standard genotoxicity testing of soluble substances, fibers bring about specific problems. Test results can be influenced by fiber dimensions, surface properties, and biopersistence. The mechanisms of fiber-induced genotoxicity are not yet clear, but direct interaction with the genetic material and indirect effects via production of reactive oxygen species (ROS) have been proposed. Asbestos did not significantly induce gene mutations in bacterial and mammalian systems but led to a clear induction of structural and numerical chromosome aberrations in cultured mammalian cells. It is the purpose of this article to critically review positive in vitro genotoxicity data obtained for asbestos in the comet assay, the chromosome aberration test, and the micronucleus test and to identify the test conditions that are specifically required for the detection of asbestos-induced genotoxicity. It is concluded that appropriate cell systems are available for testing fiber-induced genotoxicity. Fiber samples have to be well characterized and phagocytosis and cytotoxic effects have to be determined for the correct interpretation of genotoxicity test results. A combination of the micronucleus test and the comet assay using continuous treatment (without exogenous metabolic activation) seems to be well suited to detect genotoxic activity of asbestos fibers with high accuracy. Further investigations are needed to shown whether this approach can be recommended for genotoxicity testing of fibers in general.

Asbestos↗
PubMed1998

ASTM F739 method for testing the permeation resistance of protective clothing materials: critical analysis with proposed changes in procedure and test-cell design.

ASTM (American Society for Testing and Materials) Method F739-96 specifies a test-cell design and procedures for measuring the permeation resistance of chemical protective clothing. Among the specifications are open-loop collection stream flow rates of 0.050 to 0.150 L/min for a gaseous medium. At elevated temperatures the test must be maintained within 1 degree C of the set point. This article presents a critical analysis of the effect of the collection stream flow rate on the measured permeation rate and on the temperature uniformity within the test cell. Permeation tests were conducted on four polymeric glove materials with 44 solvents at 25 degrees C. Flow rates > 0.5 L/min were necessary to obtain accurate steady-state permeation rate (SSPR) values in 25 percent of the tests. At the lower flow rates the true SSPR typically was underestimated by a factor of two or less, but errors of up to 33-fold were observed. No clear relationship could be established between the need for a higher collection stream flow rate and either the vapor pressure or the permeation rate of the solvent, but test results suggest that poor mixing within the collection chamber was a contributing factor. Temperature gradients between the challenge and collection chambers and between the bottom and the top of the collection chamber increased with the water-bath temperature and the collection stream flow rate. Use of a test cell modified to permit deeper submersion reduced the gradients to < or = 0.5 degrees C. It is recommended that all SSPR measurements include verification of the adequacy of the collection stream flow rate. For testing at nonambient temperatures, the modified test cell described here could be used to ensure temperature uniformity throughout the cell.

Gloves, Protective↗
PubMed2000

A limitation of 2-stage serological testing for Lyme disease: enzyme immunoassay and immunoblot assay are not independent tests.

To improve the accuracy of testing for antibody to Borrelia burgdorferi, 2-stage conditional testing has been recommended, in which sera that yield positive or equivocal results in a first-stage test (e.g., an ELISA) are then tested by immunoblot assay. The increased specificity anticipated with sequential testing, however, depends on immunoblot assays and ELISAs being independent tests. To examine whether they are independent, control serum samples were tested with 2 different commercially available IgM ELISAs and with an IgM immunoblot assay kit. The frequency of false-positive IgM immunoblot assays was significantly higher with ELISA-reactive than with ELISA-negative serum samples (P</=.001). In addition, there was a highly significant direct correlation between the number of reactive bands on IgM blotting and the rate of false-positive results by IgM ELISA (P<.0001). These observations demonstrate that IgM ELISAs and IgM immunoblot assays for antibodies to B. burgdorferi are not independent tests. Therefore, when used in sequential testing for Lyme disease, the immunoblot assay should be considered a test that supplements rather than confirms an ELISA.

Antibodies, Bacterial↗
PubMed2005

Evaluation of PPI-0903M (T91825), a novel cephalosporin: bactericidal activity, effects of modifying in vitro testing parameters and optimization of disc diffusion tests.

OBJECTIVES: To evaluate bactericidal activity of PPI-0903M and in vitro testing parameters for this compound. METHODS: A total of 110 strains were tested to determine MIC and MBC values of PPI-0903M. MIC values were determined by reference broth microdilution whereas MBC was assessed by plating the broth from the MIC well and from those wells above the MIC for each organism onto appropriate drug-free growth media. Seventeen strains were tested by the kill-curve methodology to again evaluate the bactericidal activity of PPI-0903M. Broth microdilution results for 15 strains (in triplicate) were compared with the standard, reference method after modifying several testing parameters. Optimal disc content was assessed by testing seven reference strains with five PPI-0903M disc concentrations (5, 10, 30, 50 and 100 microg). RESULTS: PPI-0903MMBC/MIC ratios were < or = 4 in 90% of strains tested. Kill-curve studies showed >99.9% killing at concentrations < or = 4x MIC within 8 to 24 h against staphylococci and Enterobacteriaceae tested. PPI-0903M MIC results were rarely influenced by the test or medium changes evaluated [high inoculum, low calcium or reduced pH (5.0)]. Human serum had no effect on in vitro antimicrobial activity of PPI-0903M. The 10 microg disc content would be recommended for optimal correlation of MIC breakpoints of < or = 1-4 mg/L. CONCLUSIONS: PPI-0903M was generally bactericidal against tested species, and the MIC results were stable across numerous susceptibility test condition changes.

Anti-Bacterial Agents↗
PubMed2004

Dynamic testing in schizophrenia: does training change the construct validity of a test?

Dynamic testing typically involves specific interventions for a test to assess the extent to which test performance can be modified, beyond level of baseline (static) performance. This study used a dynamic version of the Wisconsin Card Sorting Test (WCST) that is based on cognitive remediation techniques within a test-training-test procedure. From results of previous studies with schizophrenia patients, we concluded that the dynamic and static versions of the WCST should have different construct validity. This hypothesis was tested by examining the patterns of correlations with measures of executive functioning, secondary verbal memory, and verbal intelligence. Results demonstrated a specific construct validity of WCST dynamic (i.e., posttest) scores as an index of problem solving (Tower of Hanoi) and secondary verbal memory and learning (Auditory Verbal Learning Test), whereas the impact of general verbal capacity and selective attention (Verbal IQ, Stroop Test) was reduced. It is concluded that the construct validity of the test changes with dynamic administration and that this difference helps to explain why the dynamic version of the WCST predicts functional outcome better than the static version.

Cognition Disorders↗
PubMed2001

Rapid D-dimer testing and pre-test clinical probability in the exclusion of deep venous thrombosis in symptomatic outpatients.

We assessed the performance of three rapid D-dimer tests (Auto Dimertest, VIDAS and Tinaquant) in combination with a pretest clinical probability model for deep venous thrombosis (DVT) in 106 consecutive outpatients with suspected DVT. Contrast venography or colour-coded duplex ultrasonography demonstrated the presence of DVT in 47 patients (14 distal DVT and 33 proximal DVT). First, we assessed the accuracy indices for different cut-off levels of the rapid D-dimer tests. Sensitivity was found to be 97.9-100%, negative predictive value (NPV) was 96.3-100%, and the exclusion rate was 24.5-31.1%. Next, the patients were grouped according to the pre-test clinical probability model in categories with low, moderate or high probability. In patients with a low pre-test probability, DVT would have been directly ruled out and the patients would not have undergone further investigations. In patients with a moderate probability, D-dimer testing and, in the case of a positive result, objective testing would have been performed and, in the case of a negative result, they would have been ruled out of having DVT. Patients with high probability would directly have undergone objective tests for DVT. The combination with the pre-test clinical probability model improved the exclusion rate (43.5-44.6%), whereas sensitivity (97.5-100%) and NPV (97.6-100%) remained roughly unchanged. The combination of rapid D-dimer tests with a pre-test clinical probability model may help to reduce unnecessary work-up in patients with suspected DVT.

Adolescent↗
PubMed1982

The effects of price information on physicians' test-ordering behavior. Ordering of diagnostic tests.

This research evaluated the effects of providing physicians with information about the prices of diagnostic tests on their subsequent test-ordering behavior. The study population consisted of 36 second- and third-year residents and 23 clinical faculty in three family practice centers affiliated with the Department of Family Medicine at Wayne State University School of Medicine, Detroit, Michigan. Study participants were asked to review four case studies, each describing ambiguous symptoms, and to indicate on an attached test order form the tests they would order for each patient. Subjects were randomly assigned either to a group that received test order forms on which to prices of diagnostic tests were printed (price-information group) or to a group that received test order forms with no prices indicated (control group). The study results show that for each of the four cases, the average number of diagnostic tests ordered was significantly lower in the price-information group than in the control group. Our findings also show an average reduction in the cumulative cost of tests ordered per patient of 31.1 per cent related to the provision of price information is discussed and reviewed in light of other approaches that have been developed to modify physician behavior in ordering diagnostic tests.

Behavior↗
PubMed1988

Pap testing intervals. Specialty differences in physicians' recommendations in relation to women's pap testing behavior.

Using data from a survey of internists, family and general practitioners, and obstetricians and gynecologists, and of women in Maryland, this article examines congruence between physicians' recommendations and women's Papanicolaou (Pap) testing behavior. Both the specialty of the physicians and the age of the women are considered. The majority of all three physician specialties recommended annual Pap tests for all their patients. However, the internists and family or general practitioners were less likely to recommend annual Pap smears for their elderly patients than for their younger patients. Obstetrician-gynecologists were consistently more likely than the other specialties to recommend annual Pap tests, to send postcards to their patients reminding them to come in for Pap tests, and to view themselves as successful in inducing their patients to come in for routine Pap testing. Both the physicians and the women reported that elderly women were less likely than younger women to receive care from obstetrician-gynecologists. Women's reported Pap testing behavior indicated that they received Pap tests with far less frequency than the physicians recommended. Older age was related to less frequent Pap testing, while having a visit with an obstetrician-gynecologist was related to increased frequency. Findings indicate the need for education of physicians about methods of improving women's attendance for Pap testing and of women about the importance of routine Pap testing.

Age Factors↗
PubMed2004

Predictive testing for Huntington's disease: relationship with partners after testing.

This study focuses on the partner relationship of tested persons, 5 years after their predictive test result for Huntington's disease (HD). We describe changes in marital status, quality of the relationship, and perceived changes in the relationship. Twenty-six carriers, 14 of their partners, 33 non-carriers, and 17 of their partners participated in the study. Qualitative and quantitative methods were used. For the majority of tested persons (about 70%), the marital status was unchanged 5 years post test. Overall, carriers rated the quality of the relationship higher than their partners did and they perceived more positive changes. Qualitative data show that a test result leading to changed roles may induce significant marital distress. Another consequence of the test may be the changes in dynamics in asymptomatic carrier couples. A pre-test discussion of the possible impact of the test result on the relationship should result in a better preparation for and more understanding of the reactions after testing. Counselling after testing should stimulate an open communication between partners with consideration of needs and anxieties of both partners.

Adult↗
PubMed1993

The detection of antisperm antibodies in serum: a comparison of the tray agglutination test, indirect immunobead test and indirect SpermCheck assay.

Testing for antisperm antibodies (ASAs) is an important part of the work-up of the sub-fertile couple, yet there is little consensus regarding the most appropriate methods. The SpermCheck assay (GSC; Bio-Rad Laboratories Inc., Diagnostics Division, Hercules, CA, U.S.A.) is supplied with wash buffer, controls and bead reagent which detects all three major classes of ASAs (IgA, IgG and IgM) in a single test. This study compared results on a bank of samples using the tray agglutination test (TAT), immunobead test (IBT), GSC and a modified SpermCheck assay to detect a single isotype in each test (SISC). The IBT and SISC showed excellent correlation, with 127/141 (90.1%) tests agreeing. There was an apparent lack of sensitivity to IgM with GSC as 8/15 (53.3%) samples testing positive with IBT and 7/15 (46.7%) testing positive with SISC were negative with GSC. Of the 24 IBT-negatives, seven (29.2%) were positive for TAT, indicating a high incidence of non-immunological agglutination, though this decreased as the TAT titre increased. The proportion of samples testing positive for IBT increased with TAT titre: 3/20 (15.0%) for TAT-negative samples, 6/10 (60.0%) for low titres and 21/24 (87.5%) for high titres. This was also observed when comparing the GSC with TAT. The TAT therefore appears useful as a first-line screen, whilst the inability of the GSC to adequately detect IgM limits its use as an indirect test. Both the IBT and SISC can be used to further investigate the type and class of ASA present.

Agglutination Tests↗
PubMed1999

Citric acid as the test meal for the 13C-urea breath test.

OBJECTIVE: Test meals are used in the urea breath test to slow gastric emptying and to increase the area of contact with the substrate. Recently, citric acid has been suggested as an improved liquid test meal. The mechanism is unknown and could act by delaying gastric emptying, decreasing the pH at the site of the bacteria, or both. Our aim was to evaluate the effects of citric acid test meals on urea hydrolysis in vivo, to identify the possible mechanism for enhanced urea hydrolysis, and to identify the minimum effective dose. METHODS: We compared the U.S. commercial 13C-urea breath test with four liquid test meals (200 ml of water) consisting of citric acid, ascorbic acid, sodium citrate, and glucose polymer and also after the subcutaneous administration of pentagastrin. We studied healthy volunteers with and without proven H. pylori infection (by serology and histology). 13C-urea was administered orally simultaneously with the liquid test meals or immediately after the pudding had been ingested. Breath samples were taken before and after oral administration of the 13C-urea. RESULTS: A dose response in urease activity was evident as the amount of citric acid was increased from 1 to 4 g. Citric acid at 1, 2, or 4 g produced significant increases in breath 13CO2 activity, compared with the commercial pudding (p < 0.05). Ascorbic acid (p = 0.053), subcutaneous pentagastrin (to lower pH) (p = 0.199), and glucose polymer (p = 0.03) (to delay gastric emptying) all approximately doubled breath 13CO2, compared with the commercial kit. Nevertheless, the increases were all significantly less than with the 4 g citric acid test meal. CONCLUSIONS: The data are consistent with the marked effect of citric acid on gastric emptying and, possibly, distribution of the urea within the stomach being largely responsible for the enhanced urease activity with citric acid test meals. It should be possible to use a low dose of citric acid (e.g., 1 g per 200 ml) to enhance the simplicity and palatability of the test.

Ascorbic Acid↗
PubMed1995

A new, peroral non-radioactive vitamin B12 absorption test compared with the Schilling test.

The results of a non-radioactive, peroral absorption test have been compared with the results of the traditional Schilling test in 31 cobalamin-deficient patients. The non-radioactive test is simple to perform, is less costly than the Schilling test and seems to give reliable results. The non-radioactive test should be performed after cobalamin treatment, but not until the plasma cobalamin value has declined to below 450 pmol/l. Normal Schilling test was noted in one-third of the patients, while normal non-radioactive test was noted in only one-fifth of the patients. The results reveal some discrepancies between the two tests regarding the response to intrinsic factor. In the non-radioactive test without intrinsic factor, the great variation in values may reflect varying secretion of intrinsic factor, possibly secondary to infestation with Helicobacter pylori. "False normal" Schilling test seems to be more common than previously believed.

Administration, Oral↗
PubMed1999

Escherichia coli ATCC 35218 as a quality control isolate for susceptibility testing of Haemophilus influenzae with haemophilus test medium.

Current National Committee for Clinical Laboratory Standards (NCCLS) susceptibility guidelines for quality control testing with Haemophilus influenzae do not include a beta-lactamase-producing strain that could detect the deterioration of the beta-lactamase inhibitor components of amoxicillin-clavulanic acid, ampicillin-sulbactam, and piperacillin-tazobactam. The objective of the study was to determine if comparable quality control results for Escherichia coli ATCC 35218, a beta-lactamase-producing strain, would be produced for the three beta-lactam-beta-lactamase inhibitor agents with Haemophilus test medium and Mueller-Hinton medium. The criteria used in this study to determine if Haemophilus test medium was acceptable for quality control testing of E. coli ATCC 35218 was that 100% of the results obtained with an antimicrobial agent-methodology combination needed to be within the acceptable NCCLS ranges established with Mueller-Hinton medium. The MIC testing results obtained by the broth microdilution and E-test methods with amoxicillin-clavulanic acid and piperacillin-tazobactam were all within the NCCLS ranges; however, the results obtained with ampicillin-sulbactam by both methods were not within the NCCLS ranges. Acceptable results were obtained by the disk diffusion methodology with ampicillin-sulbactam and piperacillin-tazobactam but not with amoxicillin-clavulanic acid. When performing susceptibility testing with H. influenzae with the beta-lactam-beta-lactamase inhibitors, in addition to quality control testing with H. influenzae ATCC 49247, testing of E. coli ATCC 35218 on Haemophilus test medium is an effective way to monitor the beta-lactamase inhibitors in some antimicrobial agent-methodology combinations.

Culture Media↗
PubMed1971

Comparison of the streptozyme test with the antistreptolysin O, antideoxyribonuclease B, and antihyaluronidase tests.

The streptozyme test was compared with the antistreptolysin O (ASO), antideoxyribonuclease B (ADN-B), and antihyaluronidase (AH) tests. One hundred and twenty-seven human serum specimens were tested by each of the four tests. The streptozyme test detected more specimens with elevated titers than any single test; however, it was not as effective as the combination of the ASO and ADN-B tests. The streptozyme test appears to be particularly useful for laboratories which rely solely on the ASO test for serological evidence of a streptococcal infection. The test can be used in these laboratories to screen all specimens with low ASO titers (<170) for the detection of ADN, AH, antinicotinamide adenine dinucleotidase, and antistreptokinase.

Animals↗
PubMed1991

Evaluation of the E test for susceptibility testing of anaerobic bacteria.

The susceptibilities of 105 clinical isolates of anaerobic bacteria were determined by a new method, the E test (AB Biodisk, Solna, Sweden), and were compared with the MICs for these organisms obtained by the reference agar dilution method by using supplemented brucella and Wilkins-Chalgren agars. The E test is a plastic strip with a predefined antibiotic gradient immobilized on one side and a MIC interpretive scale printed on the other side. Strips with cefoxitin, cefotaxime, imipenem, penicillin, metronidazole, and clindamycin were used in this study. A suspension of the test strain equal to the visual turbidity of a no. 0.5 McFarland standard was prepared and swabbed onto a 150-mm-diameter plate. The strips were applied in a radial fashion, and the plates were incubated under anaerobic conditions. After growth had occurred, an ellipse of inhibition was seen around each strip. At the point of intersection of the ellipse with the strip, the MIC was read from the interpretive scale. For most antibiotic-organism combinations, the ellipse was clear and the endpoint was sharp. The E-test MICs were interpreted after overnight and 48-h incubation for 58 of the strains. After overnight incubation, 87% of the E-test MICs were within 1 dilution of the agar dilution MICs, and 98% were within 2 dilutions. After 48 h of incubation, agreement was 86 and 97% respectively. E-test MICs obtained for the Bacteroides fragilis group after overnight incubation were more comparable than those obtained after 48 h of incubation to agar dilution MICs determined at 48 h for all drugs except clindamycin. On brucella agar, there was a 2% categorical discrepancy rate between the E-test MICs and agar dilution MICs, which occurred mostly with cefoxitin. The E test is easy to perform and read, is suitable for all anaerobes, can be used to test single patient isolates as needed, and offers the laboratory a reliable method for susceptibility testing of anaerobic bacteria.

Bacteria, Anaerobic↗
PubMed1991

Comparison of the E Test to agar dilution, broth microdilution, and agar diffusion susceptibility testing techniques by using a special challenge set of bacteria.

The E Test (AB Biodisk, Solna, Sweden) is a new method for performing antimicrobial susceptibility tests. It consists of an impervious carrier (5- by 50-mm strip) with a predefined antimicrobic gradient which is placed on an inoculated agar plate and processed like a disk diffusion test. Results are generated directly as MICs from a continuous concentration gradient covering 15 twofold dilutions, and MICs are read where the edge of the inhibition zone intersects the strip. We compared the E Test with disk diffusion, broth microdilution, and agar dilution tests by using a challenge set of 195 gram-positive and gram-negative bacteria for 14 antimicrobial agents. Also, disk diffusion, broth microdilution, and agar dilution tests were compared with each other. All test method comparisons gave greater than 94% agreement for the category of susceptibility. The E Test category agreement with disk diffusion and broth microdilution was 95.1%, and with agar dilution it was 95.2%. The E Test results were as reliable as the results obtained by the standard antimicrobial susceptibility testing methods.

Agar↗
PubMed1992

Evaluation of a monoclonal antibody-based latex agglutination test for diagnosis of cryptococcosis: comparison with two tests using polyclonal antibodies.

Cryptococcal antigen detection has become a routine biological test performed for patients with AIDS. The poor prognosis of cryptococcosis explains the need for reliable tests. We evaluated the performances of a newly commercialized agglutination test that uses a monoclonal antibody specific for cryptococcal capsular polysaccharide (Pastorex Cryptococcus; Sanofi-Diagnostics Pasteur, Marnes-la-Coquette, France) and compared them with those of tests that use polyclonal immune sera (Cryptococcal Antigen Latex Agglutination System, Meridian Diagnostics, Inc., Cincinnati, Ohio; and Crypto-LA, International Biological Labs Inc., Cranbury, N.J.). The sensitivities and specificities of the tests were compared by using purified polysaccharides and yeast suspensions. Clinical specimens (131 serum samples, 41 cerebrospinal fluid samples, 34 urine samples, and 19 bronchoalveolar lavage samples) from 87 human immunodeficiency virus-positive subjects with (40 patients) and without (47 patients) culture-proven cryptococcosis were retrospectively tested during a blinded study. The effect of pronase treatment of samples was assessed for Pastorex Cryptococcus and the Cryptococcal Antigen Latex Agglutination System, and the antigen titers were compared. Our results show that (i) during the screening, concordance among the three tests was 97%; (ii) the use of pronase enhanced both the sensitivities and specificities of the Pastorex Cryptococcus test; (iii) titers agreed for 67% of the cerebrospinal fluid samples and 60% of the serum samples; and (iv) cryptococcosis was detected equally well with Pastorex Cryptococcus and with the other tests, whatever the infecting serotype (A, B, or D). The meaning of in vitro sensitivity and the relationship between titers and sensitivity are discussed. The results show that Pastorex Cryptococcus is a rapid and reliable test for the detection of cryptococcal antigen in body fluids and suggest that kits cannot be used interchangeably to monitor antigen titers in patients.

AIDS-Related Opportunistic Infections↗
PubMed1993

E test as susceptibility test and epidemiologic tool for evaluation of Neisseria meningitidis isolates.

The E test (AB Biodisk, Solna, Sweden), a new approach developed to test antimicrobial susceptibility, was compared with the agar dilution method for seven-drug antibiogram analysis of Neisseria meningitidis isolates. The overall E-test quantitative accuracy (+/- 1 log2 dilution) was 93% compared with that of agar dilution testing. The E test was then used to perform the susceptibility tests on a 10-year sample of 102 N. meningitidis isolates, including 5 from a recent epidemic outbreak in the University of Iowa (Iowa City) community. The E test proved to be an efficient methodology for identifying common source clusters of meningococcal disease having resistance to rifampin or sulfonamides. Moreover, the data demonstrated a recent increase in penicillin MICs (MIC for 90% of strains, 0.094 microgram/ml) and an escalation of high-level resistance to trimethoprimsulfamethoxazole (33%) and rifampin (14%). The E test should be considered a simple and accurate susceptibility method for the emerging need to test meningococci and other pathogenic neisserias. Chocolate Mueller-Hinton agar was observed to provide the best support of growth and E-test MIC results that correlated well with results of the reference agar dilution method previously used for neisserias.

Agar↗