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PubMedDate not supplied

PCR-based detection and typing of human adenoviruses in clinical samples.

Since human adenoviruses (Ad) are associated with a variety of diseases, there is need for a fast and sensitive diagnostic procedure. The polymerase chain reaction (PCR) has been previously applied for the detection and typing of adenoviruses directly in clinical samples. So far, only Ad8, Ad31, Ad40 and Ad41 could be typed by PCR. To extend the technique of type-specific PCR to other adenovirus serotypes, type-specific primers for Ad1, Ad2, Ad4, Ad5, Ad19 and Ad37 were evaluated. In the present study, 50 stool and 68 eye swab specimens were first tested for the presence of adenoviruses using genus-specific primers. Adenoviruses could be detected in 42 stool and 47 eye swab samples. While the adenovirus-positive stool samples were subsequently typed with primers for Ad2, Ad5, Ad31, Ad40 and Ad41, the positive eye swab specimens were typed with primers for Ad4, Ad8, Ad19 and Ad37.

Adenovirus Infections, Human↗
PubMed2003

Phenotypic description and antimicrobial susceptibilities of Aerococcus sanguinicola isolates from human clinical samples.

This report describes the clinical sources and phenotypic characterization of 16 isolates of Aerococcus sanguinicola. Sixteen conventional tests were used to describe and differentiate the 16 isolates of A. sanguinicola from 30 strains of Aerococcus viridans, 27 strains of Aerococcus urinae, and a single strain each of Aerococcus christensenii and Aerococcus urinaehominis. The phenotypic characterizations of the type strains for each species and 14 A. sanguinicola isolates were also compared in the two reference laboratories. A. sanguinicola are catalase-negative, vancomycin-susceptible, gram-positive cocci arranged in clusters and tetrads, as are all Aerococcus species except A. christensenii (which is arranged in short chains). All 16 isolates of A. sanguinicola were leucine aminopeptidase and pyrrolidonylarylamidase positive, which is unique to this species among the aerococci. All A. sanguinicola isolates grew in broth containing 6.5% NaCl, hydrolyzed hippurate, and were variable in the bile-esculin test. None of the isolates deaminated arginine or were Voges-Proskauer positive. The type strain of A. sanguinicola was isolated from a blood culture of a patient living in Denmark. Seven additional isolates were from patients living in Canada, all with urinary tract infections (six were female). Eight isolates were from patients living in five different states in the United States; five were from patients with urinary tract infections, and three were from blood cultures of one patient each with pneumonia, suspected endocarditis, and unknown clinical conditions. The antimicrobial susceptibility patterns were unremarkable; all isolates tested were susceptible to penicillin, amoxicillin, cefotaxime, cefuroxime, erythromycin, chloramphenicol, vancomycin, quinupristin-dalfopristin (Synercid), rifampin, linezolid, and tetracycline. Six of the 15 cultures were resistant to ciprofloxacin and levofloxacin, but all 15 strains were susceptible to sparfloxacin. High-level resistance was detected for meropenem (2 strains) and trimethoprim-sulfamethonazole (1 strain). Intermediate resistance was detected for trimethoprim-sulfamethoxazole (10 strains) and clindamycin (3 strains).

Anti-Bacterial Agents↗
PubMed2000

Estimation of Wechsler Adult Intelligence Scale-III index scores with the 7-subtest short form in a clinical sample.

A 7-subtest short form of the Wechsler Adult Intelligence Scale-III (WAIS-III) previously demonstrated good comparability in estimating Full Scale and Verbal IQ summary scores, with adequate comparability in estimating Performance IQ. In a mixed clinical sample of 295 patients, the current study assessed the equivalence of the index scores generated from the full and prorated WAIS-III. The results revealed correlations corrected for redundancy of .90, .86, .87, and .75 for the Verbal Comprehension (VCI), Perceptual Organization (POI), Working Memory (WMI), and Processing Speed (PSI) indexes, respectively. Although the 7-subtest short form of the WAIS-III was not designed to estimate index scores, adequate estimates are viable for VCI, POI, and WMI when the goal is to obtain group, rather than individual, data points.

Adult↗
PubMedDate not supplied

Detection of hepatitis A antibodies by ELISA using saliva as clinical samples.

The possibility of detecting acute infection and immunity using body fluids that are easier to collect than blood, mainly in children, would facilitate the investigation and follow-up of outbreaks of hepatitis A (HAV). Our study was carried out to evaluate the detection of anti-HAV IgM, IgA and total antibodies in saliva using serum samples as reference. Forty three paired serum and saliva samples were analyzed. From this total, 24 samples were obtained from children and 1 from one adult during the course of acute hepatitis A; an additional 18 samples were obtained from health professionals from Adolfo Lutz Institute. The sensitivity to detect anti-HAV IgM was 100% (95%CI: 79.1 to 100.0%), employing saliva as clinical samples. In detecting anti-HAV IgA, the sensitivity was 80. 8% (95%CI: 60.0 to 92.7%) and for the total antibodies was 82.1% (95%CI: 62.4 to 93.2%). The specificity was 100% for each. The rate of agreement was high comparing the results of serum and saliva samples for detecting HAV antibodies. We conclude that saliva is an acceptable alternative specimen for diagnosing acute hepatitis A infection, and for screening individuals to receive hepatitis A vaccine or immunoglobulin.

Adult↗
PubMed2006

Symptoms of traumatic stress and substance use in a non-clinical sample of young adults.

The link between symptoms of traumatic stress and substance abuse is well established in the literature, but the specific features of trauma that contribute to substance abuse remain elusive and have not been studied in non-clinical samples. The current study assessed the relationship between traumatic symptoms and alcohol use in a sample of Midwestern college students. Using multiple regression analysis, traumatic stress symptoms, overall, accounted for 55% of the variance in use of alcohol. Most importantly, dissociative behavior, intrusive thoughts, as well as low levels of tension reduction behaviors uniquely contributed to the self-reported use of alcohol. We discuss the importance of educating college-aged students about this relationship as a preventative measure to alcohol and other drug use/abuse.

Adaptation, Psychological↗
PubMed1993

Nested polymerase chain reaction for detection of Mycobacterium tuberculosis in clinical samples.

The nested polymerase chain reaction technique was compared with the conventional smear and culture methods for detection of Mycobacterium tuberculosis. The nested polymerase chain reaction used in this study showed excellent specificity, sensitivity, and agreement with the conventional methods for 417 clinical samples, indicating a contribution to the rapid diagnosis of mycobacterial infectious diseases.

Base Sequence↗
PubMed2003

Quality control trial for human immunodeficiency virus type 1 drug resistance testing using clinical samples reveals problems with detecting minority species and interpretation of test results.

Between January and March 2000, a quality control panel for human immunodeficiency virus (HIV) drug resistance testing was analyzed by 20 laboratories in five countries. The panel consisted of three clinical samples with different drug resistance genotypes and phenotypes and one HIV-negative plasma. Participants were asked to report the methods used for amplification and sequencing, a list of drug resistance-associated mutations that were detected in the protease and reverse transcriptase of each sample, and an interpretation concerning the susceptibility or resistance to 14 antiretroviral drugs. A total of 22 genotypic data sets were generated, which showed an overall good technical quality except for three participants, who failed to report key mutations for drug resistance. Problems were encountered in three respects: (i). resistant minorities of L90M in the protease, which were determined to about 12% by real-time amplification, were only detected by one-fourth of the participants; (ii). newly described resistance mutations were frequently not reported; and (iii). interpretations of drug resistance-associated mutations varied widely, in particular for protease inhibitors. In some cases, different interpretations were caused by differences in the detection of resistant minorities, but even for the same genotypic profile, interpretations varied considerably. Similar discrepancies were revealed if current Web-based interpretation systems were used to predict drug resistance for samples of the proficiency panel. This indicates that a consensus for the interpretation of drug resistance-associated mutations is urgently needed.

Antiviral Agents↗
PubMed2003

The Depression Anxiety Stress Scales (DASS): normative data and latent structure in a large non-clinical sample.

OBJECTIVES: To provide UK normative data for the Depression Anxiety and Stress Scale (DASS) and test its convergent, discriminant and construct validity. DESIGN: Cross-sectional, correlational and confirmatory factor analysis (CFA). METHODS: The DASS was administered to a non-clinical sample, broadly representative of the general adult UK population (N = 1,771) in terms of demographic variables. Competing models of the latent structure of the DASS were derived from theoretical and empirical sources and evaluated using confirmatory factor analysis. Correlational analysis was used to determine the influence of demographic variables on DASS scores. The convergent and discriminant validity of the measure was examined through correlating the measure with two other measures of depression and anxiety (the HADS and the sAD), and a measure of positive and negative affectivity (the PANAS). RESULTS: The best fitting model (CFI =.93) of the latent structure of the DASS consisted of three correlated factors corresponding to the depression, anxiety and stress scales with correlated error permitted between items comprising the DASS subscales. Demographic variables had only very modest influences on DASS scores. The reliability of the DASS was excellent, and the measure possessed adequate convergent and discriminant validity Conclusions: The DASS is a reliable and valid measure of the constructs it was intended to assess. The utility of this measure for UK clinicians is enhanced by the provision of large sample normative data.

Adolescent↗
PubMed2003

Molecular diagnosis of leishmaniosis in dogs. Comparative application of traditional diagnostic methods and the proposed assay on clinical samples.

Leishmaniosis is a zoonotic, parasitic disease caused by members of the genus Leishmania. The disadvantages of the traditional methods have currently rendered the polymerase chain reaction (PCR), the most reliable alternative for the laboratory diagnosis of this disease. Several relevant protocols have been described in the past but their application is in most cases limited to research use. The latter combined with the diagnostic problems that can be caused by the genetic variability of the different Leishmania strains or the presence of PCR inhibitors, indicate that an alternative approach should be followed for the development of a standard diagnostic tool for leishmaniosis. In the present study, we have evaluated several PCR-based protocols, in order to identify a primer combination that would allow the reliable detection of Leishmania DNA from clinical material and the verification of its results, in a manner that could be applicable even for routine use. The evaluation consisted of a BLAST verification of the specificity of the previously described primers, PCR testing, and optimisation of the reaction conditions. Our assessment was completed with the comparative evaluation of the results produced by the proposed PCR assay, light microscopy, and indirect fluorescent antibody technique (IFAT), on clinical samples collected from dogs suspected of leishmaniosis. The proposed assay which consists of a combination of two pairs of primers, targeted to different areas of the kinetoplast DNA of Leishmania spp., specific for Leishmania infantum, Leishmania donovani and Leishmania chagasi, showed optimum performance on our test samples, and detected 41.9% Leishmania-positive dogs from our 160 clinical cases. From the same number of cases, 46.25% were positive by IFAT (titre > or =200), and 19% by microscopic examination of lymph node aspirates.

Animals↗
PubMed1998

Effect of genotypes on the quantification of hepatitis C virus (HCV) RNA in clinical samples using the Amplicor HCV Monitor Test and the Quantiplex HCV RNA 2.0 assay (bDNA).

The Amplicor HCV Monitor test and the Quantiplex HCV RNA 2.0 (bDNA) assay are two commercially available assays for the quantification of hepatitis C virus (HCV) RNA in clinical samples. A direct comparison of the two assays was carried out using sera frozen previously from patients known to be chronically infected with HCV. Overall, 61 samples from 51 patients were tested simultaneously by the two methods: 67% (28/42) of the patients were infected by HCV genotype/serotype 1, 10% (4/42) with type 2, and 24% (10/42) with type 3. When the absolute value from each assay was examined, the Quantiplex assay gave a consistently higher reading and the mean logarithmic difference between the two assays was 1.4 (1.0 in type 1, 2.0 in type 2, and 2.2 in type 3). When analyzed according to genotype, strong correlation was observed between the two assays for type 1 (r = 0.83, 95% CI 0.63-0.93, P < 0.01), but not for nontype 1 samples. Despite the difference in absolute level reported by the two assays, there was a consistent trend of change in HCV RNA concentration by both assays in patients whose consecutive samples were analyzed and the differences between the two assays in consecutive samples were within 0.4 log of each other. The results suggested that with samples containing genotype 1, the Amplicor assay was more sensitive than the Quantiplex assay by about one log. However, the sensitivities of the two assays with nontype 1 samples were much closer probably due to the failure of the Amplicor assay to quantify nontype 1 genotypes effectively.

Adolescent↗
PubMed1989

The Social Phobia and Anxiety Inventory: concurrent validity with a clinic sample.

The Social Phobia and Anxiety Inventory (SPAI) is an empirically derived self-report inventory developed as a specific measure of social phobia. The current investigation included two studies. The first examined the correlation of the SPAI with daily social behavior of a clinic sample of social phobics. The results indicated that the SPAI provides a reasonable indication of the distress experienced during daily social encounters in three dimensions: behavior, cognitions, and overall distress. The second study examined the validity of the SPAI with reference to the somatic response and avoidance behavior of social phobics. The results indicated that the somatic items of the SPAI are related to the somatic response of social phobics and that performance on the SPAI is associated with avoidance behavior in an anxiety-producing task.

Adult↗
PubMed2005

Evaluation of a single-tube fluorogenic RT-PCR assay for detection of bovine respiratory syncytial virus in clinical samples.

Bovine respiratory syncytial virus (BRSV) causes severe disease in naive cattle of all ages and is a common pathogen in the respiratory disease complex of calves. Simplified methods for rapid BRSV diagnosis would encourage sampling during outbreaks and would consequently lead to an extended understanding of the virus. In this study, a BRSV fluorogenic reverse transcription PCR (fRT-PCR) assay, based on TaqMan principle, was developed and evaluated on a large number of clinical samples, representing various cases of natural and experimental BRSV infections. By using a single-step closed-tube format, the turn-around time was shortened drastically and results were obtained with minimal risk for cross-contamination. According to comparative analyses, the detection limit of the fRT-PCR was on the same level as that of a nested PCR and the sensitivity relatively higher than that of a conventional PCR, antigen ELISA (Ag-ELISA) and virus isolation (VI). Interspersed negative control samples, samples from healthy animals and eight symptomatically or genetically related viruses were all negative, confirming a high specificity of the assay. Taken together, the data indicated that the fRT-PCR assay can be applied to routine virus detection in clinical specimens and provides a rapid and valuable tool in BRSV research.

Animals↗
PubMed2000

Multilocus enzyme electrophoresis analysis of Aspergillus fumigatus strains isolated from the first clinical sample from patients with invasive aspergillosis. EBGA Network. European Research Group on Biotype and Genotype of Aspergillus.

The genotypes of 50 isolates of Aspergillus fumigatus from 11 patients with invasive aspergillosis, obtained from three hospitals in different geographical areas, were determined by multilocus enzyme electrophoresis (MLEE). The study analysed the genetic polymorphism of multiple isolates from the first sample. Seven of the 14 enzymic loci studied were polymorphic, giving rise to eight different electrophoretic types. For nine of 11 patients studied, no polymorphism was observed in isolates within the first clinical sample. Analysis of genetic distance between electrophoretic types demonstrated a genetic heterogeneity within each geographical site. Moreover, some genotypes were preferentially found in a given area and this revealed a population structure within these geographical sites. Therefore, the epidemiology of A. fumigatus should be considered separately for each of these areas. The multiple discriminatory markers of MLEE seem to provide a powerful tool for increasing the understanding of the biology of this fungus.

Alleles↗
PubMed1999

[Questionnaire of self-efficacy, optimism and pessimism: reconstruction, selection of items and validation of an instrument by means of examinations of clinical samples].

Self-efficacy and optimism are considered highly useful parameters in the context of therapeutic indication prognosis, and outcome evaluation. In our study, a combined questionnaire measuring "self-efficacy" (Schwarzer u. Jerusalem 1986) and "optimism" (Scheier u. Carver 1985) was restructured and reduced in length. The resulting instrument was compared with its original version in five different clinical samples comprising n = 726 inpatients. Factor analyses achieved the bidimensionality in the construct "optimism" as postulated by Scheier and Carver, this yielding the two factors "optimism" and "pessimism". The statistical validation resulted in good to optimal values. Factor analyses considering the different clinical groups showed that certain items failed to be invariably approprieted to one factor. Therefore, we decided in favor of an item reduction, resulting in half the original number: 5 items representing "self-efficacy" 2 for "optimism" and 2 for "pessimism". The scale values of the reduced version showed no relevant deviations compared to the original, and hence we conclude that the reduction does not result in a loss of information.

Humans↗
PubMed1987

[Susceptibility of some bacteria isolated from clinical samples to third-generation cephalosporins].

In this study, we evaluated the susceptibility of some Staph. aureus, Staph. epidermidis, E. coli, Klebsiella, Proteus, Pseudomonas strains to third-generation cephalosporins in-vitro by disk diffusion technique which are in use in Turkey. These bacteria isolated from various clinical samples at the bacteriology laboratory in Ibni Sina hospital between January 1986-January 1987. The antibiotics used in this study are ceftizoxime, cefotaxime, ceftriaxone, cefoperazone. According to the results of this study, it can be said that cefotaxime is the most effective third-generation cephalosporin against all these bacteria by testing in-vitro.

Bacteria↗
PubMed1987

Measurement of terminal deoxynucleotidyl transferase mRNA in clinical samples: a new parameter in analysis of leukemia cells.

A 1750 base pair cDNA to human terminal deoxynucleotidyl transferase (TdT) has been cloned. This cDNA detects a dominant 2200 base pair messenger RNA species in normal and leukemic cells synthesizing the enzyme. A quantitative dot blot assay was utilized to survey a number of clinical samples from patients with TdT positive and negative leukemias as well as cells from normal volunteers. A linear relationship was detected between the amount of TdT mRNA and the amount of enzyme activity in bone marrow cells. The assay is sensitive enough to detect normal TdT levels in bone marrow, and distinguish these levels from the lack of such mRNA in peripheral blood and bone marrow of patients with myeloid leukemia. Elevated levels of mRNA were found in two cases of patients in clinical remission. The prognostic significance of these observations must await further study.

Base Composition↗
PubMed2006

American tegumentary leishmaniasis: direct species identification of Leishmania in non-invasive clinical samples.

Species identification is highly relevant for improved prognosis and adequate treatment of American tegumentary leishmaniasis (ATL). PCR-based methods are available for this purpose but should be simplified to improve accessibility. As a first step in this process, this paper describes a simplified protocol for collection of clinical samples. Using samples from 44 Bolivian patients with confirmed ATL, we demonstrated that hsp70 PCR-RFLP on skin scrapings collected with a tooth pick allowed identification of the parasite species with a sensitivity of 95% and specificity of 100%. Our method should greatly facilitate individual patient management and epidemiological surveillance of ATL.

Animals↗
PubMed1991

Methicillin-resistant staphylococcal strains isolated from clinical samples.

Infections caused by methicillin-resistant Staphylococcus strains (MRSS) have become an increasing problem both as community-acquired and nosocomial infections. In order to eradicate colonization as well as to cure infections, optimal antibiotic treatment is required. In this study we examined the incidence of MRSS in clinical samples and compared the antibiotic susceptibility pattern of methicillin-resistant Staphylococcus aureus with that of methicillin-resistant Staphylococcus non-aureus strains. All the MRSS were resistant to penicillin. Among them there was a variation in the percentage of strains resistant to various antimicrobial agents. Within MRSS the most frequent resistances were those to erythromycin and norfloxacin. Ciprofloxacin and teicoplanin were the most effective antibiotics tested against MRSS, followed in activity by vancomycin and imipenem. The incidence of antibiotic resistance among MRSS was significantly higher than that among methicillin-susceptibility staphylococcal species.

Cross Infection↗