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PubMed2005

Layered peptide arrays: high-throughput antibody screening of clinical samples.

High-throughput methods to detect and quantify antibodies in sera and other patient specimens have use for many clinical and laboratory studies, including those associated with cancer detection, microbial exposures, and autoimmune diseases. We developed a new technique, termed layered peptide array (LPA), to serve as a screening tool to detect antibodies in a highly multiplexed format. We demonstrate here that a prototype LPA was capable of producing approximately 5000 measurements per experiment and appeared to be scalable to higher throughput levels. Sera and saliva from Sjögren's syndrome patients served as a test set to examine antibody titers in clinical samples. The LPA platform exhibited both a high sensitivity (100%) and high specificity (94%) for correctly identifying SSB antigen-positive samples. The multiplex capability of the platform was also confirmed when serum and saliva samples were analyzed for antibody reactivity to several peptides, including Sjögren's syndrome antigens A and B. The data indicate that LPA analysis will be a useful method for a number of screening applications.

Antibodies↗
PubMed2001

Cross-cultural comparison of clinical samples of Norwegian and Iowan women with substance-related disorders.

The purpose of this study was to compare two clinical samples of women substance abusers, one from Iowa and one from Norway, along several dimensions: "substance abuse" history, current "abuse" pattern, parental and spouse drinking history, symptoms of psychological distress, and "vulnerability" to relapse. A higher percentage of women substance abusers from Iowa were divorced and lived alone, were "vulnerable" to relapse, and reported more life problems secondary to their substance use than women from Norway. The Iowa women had used more substances in addition to alcohol than had the Norwegian women. The Norwegian women reported greater intensity of emotional distress than the women from Iowa.

Adolescent↗
PubMed2004

In vitro antibacterial activity of the peptide PsVP-10 against antimicrobial-resistant Enterococcus faecalis isolated from clinical samples.

OBJECTIVE: To analyse the antimicrobial activity of the peptide PsVP-10 against 67 resistant Enterococcus faecalis strains isolated from clinical samples. METHODS: The qualitative disc diffusion method and MIC determinations were used. RESULTS: The presence of several multidrug-resistant phenotypes of E. faecalis was demonstrated, in which there were high MICs to chloramphenicol, tetracycline, vancomycin, cefaloridine, ampicillin and gentamicin. In comparison, the peptide PsVP-10 showed lower MICs against all the multidrug-resistant and susceptible E. faecalis. CONCLUSIONS: There is an urgent need for the development of novel antimicrobial agents against the highly resistant E. faecalis. The present study shows that the peptide PsVP-10 might make a contribution to the solution of this serious problem.

Anti-Bacterial Agents↗
PubMed2004

Novel method using a laser scanning cytometer for detection of mycobacteria in clinical samples.

In order to evaluate the capacity of laser scanning cytometry (LSC) to detect acid-fast bacilli directly on clinical samples, a comparison between Kinyoun-stained smears analyzed under light microscopy and propidium iodide-auramine-stained smears analyzed by LSC was performed. The results were compared with those for culture on BACTEC MGIT 960. LSC is a new, reliable methodology to detect MYCOBACTERIA:

Cytophotometry↗
PubMed1996

The sub-typing of Listeria monocytogenes isolates from food, environments surrounding food manufacturing sites, and clinical samples in New Zealand using multilocus enzyme electrophoresis.

Listeria monocytogenes strains isolated from food (cheese, meat and fish), environments surrounding food manufacturing sites, and clinical samples from throughout New Zealand were analyzed by multilocus enzyme electrophoresis. One hundred and twenty four strains were divided into 56 electrophoretic types, containing primarily clinical isolates and environmental isolates.

Bacterial Typing Techniques↗
PubMed2001

Diagnosis of foot-and-mouth disease by RT-PCR: use of phylogenetic data to evaluate primers for the typing of viral RNA in clinical samples.

The results of type-specific RT-PCR diagnostic assays on foot-and-mouth disease (FMD) viruses in clinical samples were mapped onto serotype-specific dendrograms representing the degree of nucleotide sequence variation between the FMD virus isolates. This novel approach assisted the selection of suitable PCR primer sets for the diagnosis of FMD virus isolates belonging to different topotypes within each serotype. These interpretations were qualified by using a universal (FMD virus group) specific primer to confirm that FMD virus RNA had been extracted from the samples under investigation. The analyses showed that the design of primer sets for the detection of FMD virus serotypes O, A, Asia 1, SAT 1 and SAT 3 were generally satisfactory, as most virus isolates within the major virus sub-groupings were successfully detected. However, the FMD virus serotype C and SAT 2 specific primers were less efficient as certain virus sub-groups were not detected. This identified the need for additional or alternative primers to improve RT-PCR procedures for more comprehensive detection of divergent virus strains within these serotypes. There were some examples where not all virus isolates from the same outbreak reacted with particular type-specific primers which suggested that either further minor refinements may be necessary in the primer design or that there were shortcomings in the RT-PCR methodology.

Animals↗
PubMed2005

The relationship between eating self-efficacy and eating disorder symptoms in a non-clinical sample.

Most research on eating self-efficacy has focused on its relationship with eating behaviors and weight-loss in clinical populations. The purpose of this study was to investigate the relationship between eating self-efficacy and the behavioral and psychological characteristics associated with eating disorders in a non-clinical sample of adults. A total of 219 men and women aged 18 and older completed questionnaires measuring eating disorder symptoms and eating self-efficacy. The results indicated that low confidence in the ability to control eating while experiencing negative emotions was associated with greater weight preoccupation and bulimic thought and behaviors. In addition, low confidence in ability to control eating when an abundance of food is available was inversely related to feelings of ineffectiveness or general negative self-evaluation. Ultimately, the findings suggest that low eating self-efficacy may also be associated with eating problems within populations not seeking treatment for either eating disorders or weight-loss. The implications of the findings are discussed.

Adolescent↗
PubMed1995

Conduct disorder and substance use disorder: comorbidity in a clinical sample of preadolescents and adolescents.

OBJECTIVE: To examine the rate of comorbidity between conduct disorder and substance use disorder in a clinical sample using the Diagnostic Interview for Children and Adolescents--Revised. METHOD: Examined the pattern of conduct disorder symptoms, including type, number, and severity, in conduct-disordered youth diagnosed with, and without a comorbid substance use disorder. RESULTS: The examination revealed no significant differences in the incidence of comorbidity between younger (aged 10 to 13) and older (above age 13) youth. Among youth who met criteria for conduct disorder, 52% also met criteria for a substance use disorder. Odds ratios indicated that the probability of comorbidity of conduct and substance use disorders was higher in the younger group. CONCLUSION: Substance abuse and dependence tend to develop rapidly following first use, suggesting that a slim window of opportunity exists to prevent substance disorders once drug use has begun.

Adolescent↗
PubMed1991

Rapid monitoring of benzodiazepines in clinical samples by using on-line column switching HPLC.

We have determined routinely administered benzodiazepines and some of their metabolites in clinical samples by using on-line column switching for sample clean-up and reversed-phase high-pressure liquid chromatography (HPLC). The plasma sample is directly injected onto a cyanopropyl (CN) pre-column, washed with buffer, and eluted onto an analytical column (C18, octadecylsilane). Analytical recovery from drug-supplemented plasma samples was greater than 85% for all the benzodiazepines tested. The response of the analytical column varied linearly with drug concentrations over the range 0.01-1 mg/L in plasma, with CVs ranging from 0.3% to 1.8% within-day and 5.5% to 15.7% between days. The detection limit (signal-to-noise ratio greater than 3) was 0.01 mg/L (1 ng on column) for all of the benzodiazepines and their metabolites. The advantages of the technique include the extreme cleanliness of the traces, no requirement for off-line sample preparation, and ease of automation.

Autoanalysis↗
PubMed2001

Evaluation of a real-time PCR assay for detection of Bordetella pertussis and B. parapertussis in clinical samples.

A real-time PCR assay based on the TaqMan technology was developed for the detection of Bordetella pertussis and B. parapertussis in clinical samples. The assay was evaluated with 182 specimens from 153 patients with and without symptoms of pertussis. The analytical sensitivity ranged from 0.1 to 10 cfu for B. pertussis and B. parapertussis, respectively, and diagnostic sensitivity was 94.1% when culture was used as a reference. No sample from a patient without symptoms of pertussis was positive in PCR. Twenty-four of 28 patients who were negative by culture and positive by PCR assay met the CDC clinical case definition for pertussis; the remaining four patients had paroxysms of shorter duration. Intra- and inter-assay variation were <5% and results were available within 4 h.

Adolescent↗
PubMed2006

The OCI-R: validation of the subscales in a clinical sample.

The psychometric properties of the Obsessive-Compulsive Inventory-Revised (OCI-R) subscales have not been validated in a clinical sample of individuals diagnosed with obsessive-compulsive disorder (OCD). Data were collected on 186 patients diagnosed with OCD and 17 patients diagnosed with generalized anxiety disorder (GAD) using the OCI-R and the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS). Confirmatory factor analysis revealed an acceptable factor structure. Patients with a given primary symptom subtype were elevated on the corresponding subscale of the OCI-R compared to other OCD patients and patients with GAD. In addition, patients who acknowledged that symptom subtype as present but not primary on the Y-BOCS had elevated OCI-R scores on that scale compared to patients who did not endorse that symptom subtype and patients with GAD. Results indicate that the subscales of the OCI-R are valid measures of six symptom subtypes of OCD. The OCI-R is a psychometrically sound, brief instrument. The current data combined with previous efforts suggest that it is appropriate for clinical and non-clinical populations, and for clinical and research purposes. Further research should examine the sensitivity of the specific subscales to treatment effects, and the potential for adding more items to account for other symptom domains of OCD.

Adult↗
PubMed2003

Classification of Ralstonia pickettii-like isolates from the environment and clinical samples as Ralstonia insidiosa sp. nov.

Thirteen Ralstonia pickettii-like isolates from the environment (water, soil and activated sludge) and human clinical samples (including respiratory secretions of cystic fibrosis patients) were investigated in a polyphasic taxonomic study that employed 16S rDNA sequence analysis, DNA-DNA hybridization, determination of DNA base composition, whole-cell protein analysis, biochemical characterization and PCR-based assays. All isolates were classified as a novel Ralstonia species, for which the name Ralstonia insidiosa sp. nov. is proposed. The type strain, LMG 21421T (= CCUG 46789T), was isolated from the sputum of a patient with acute lymphoblastic leukaemia. R. insidiosa can be differentiated from other species of the genus Ralstonia and phenotypically similar species (including the Burkholderia cepacia complex and Achromobacter xylosoxidans) by a variety of biochemical tests, whole-cell protein analysis and several PCR-based assays. Some outstanding issues in the taxonomy of the genus Ralstonia are also discussed.

Base Composition↗
PubMed1992

Detection of Mycoplasma pneumoniae in clinical samples from pediatric patients by polymerase chain reaction.

The polymerase chain reaction (PCR) technique was used to detect Mycoplasma pneumoniae DNA in clinical samples (nasopharyngeal aspirations or bronchoalveolar lavages) obtained from 100 children, 1 month to 16 years old. PCR allowed the detection of M. pneumoniae DNA from 20 out of the 100 patients studied. In 16 cases, PCR positivity was associated with acute respiratory symptomatology. For five PCR-positive patients, a positive culture or a serological response evidenced acute M. pneumoniae infections. A lack of antibody response was observed particularly with immunocompromised children and infants less than 12 months old. The amount of M. pneumoniae DNA in the PCR was estimated in a semiquantitative way by comparison of its hybridization signal with those obtained for 100, 10, and 1 color-changing unit (CCU) of the M. pneumoniae FH strain. Small amounts (less than or equal to 10(2) CCU/ml) of M. pneumoniae were found in samples from asymptomatic patients, while larger amounts (greater than or equal to 10(2) to greater than or equal to 10(4) CCU/ml) were found for 8 out of 10 patients with acute pneumonia.

Adolescent↗
PubMed1997

Alexithymia in a clinical sample of obese women.

The purpose was to examine the rate of alexithymia as measured by the Toronto Alexithymia Scale in a clinical sample of obese women without Binge Eating Disorder. Subjects included 165 inpatients in an eating disorder unit and 135 normal-weight individuals (comparison group). The obese subjects also completed a series of tests including the Minnesota Multiphasic Personality Inventory, the Anxiety Scale Questionnaire, and the Eating Attitude Test. Analysis showed that the subgroups of obese persons differ in important ways and cannot necessarily be treated, studied, or understood through a single paradigm. Alexithymic behaviour, too, was not characteristic of obesity, but it was present only in the subgroup of subjects with psychopathological characteristics.

Adult↗
PubMed1996

Relationships of the WISC-R and K-BIT for an adolescent clinical sample.

Correlations between the Wechsler Intelligence Scale for Children-Revised IQs and the Kaufman Brief Intelligence Test were determined for a clinical sample of 44 adolescents (35 with learning disabilities, 9 with mental retardation) who underwent routine three-year reevaluations. Even though three years had elapsed between the WISC-R and K-BIT scores, the correlations among the total and subscale scores of these measures were high and were all statistically significant. A significant mean difference was found between the WISC-R Verbal IQ and its K-Bit counterpart, the Vocabulary subscales. Implications of findings for assessment specialists are discussed.

Adolescent↗
PubMed2005

A refined long RT-PCR technique to amplify complete viral RNA genome sequences from clinical samples: application to a novel hepatitis C virus variant of genotype 6.

The goal of this study was to adapt a long RT-PCR technique to amplify large PCR fragments from the genome of hepatitis C virus (HCV) isolates using clinical samples. This was done by using a reverse transcriptase devoid of RNase H activity and a mixture of two antibody-bound thermostable polymerases to combine the high processivity of Taq and the high fidelity of Pwo with its 3'-->5' exonuclease activity. Other modifications included gentle handling during RNA extraction, the absence of tRNA and random primers, a two-step reverse transcription procedure to optimize cDNA synthesis, and increasing the annealing temperature for primers. With this approach, the HCV-1 genome (nucleotides 35-9282) was amplified consistently as two overlapping fragments of 5344 and 4675 bp from a pooled chimpanzee plasma sample containing approximately 10(6) genome copies of HCV RNA/ml. Using the conditions that we identified, 96% of the complete genomic sequence of a distinct HCV genotype 6 variant (km45) was determined from less than 300 microl of serum. This method should prove useful for molecular, epidemiological and clinical studies of hepatitis C where samples are limited but complete virus sequence is required, for example, identifying mutational hot spots of HCV under specific clinical conditions.

Animals↗
PubMed2003

Quantitative real-time PCR for detection of monkey B virus (Cercopithecine herpesvirus 1) in clinical samples.

A TaqMan based real-time PCR assay was developed for rapid detection and quantitation of herpes B virus (Cercopithecine herpesvirus 1) in clinical samples. The assay utilizes B virus-specific primers and a probe to the non-conserved region of the gG gene to discriminate B virus from closely related alphaherpesviruses. Fifty copies of B virus DNA could be detected with 100% sensitivity with a wide range of quantitation spanning 6 logs. The assay was highly reproducible with intra- and inter-assay coefficients of variation of 0.6 and 2.4%, respectively. Clinical utility of the developed real-time PCR was evaluated by testing genomic DNA prepared from B virus clinical isolates (n=23) and human and monkey clinical specimens (n=62). This novel method was also compared with conventional cell culture with respect to sensitivity and specificity. TaqMan PCR assay was shown to be equally specific and more sensitive than culture method (culture vs. PCR sensitivity 50%) and was able to identify all B virus clinical isolates tested. Fast, reliable assessment of B virus DNA in infected cells and tissues makes real-time PCR assay a valuable tool for diagnosis and management of B virus infections.

Animals↗
PubMed2006

The appraisal of social concerns scale: psychometric validation with a clinical sample of patients with social anxiety disorder.

The Appraisal of Social Concerns (ASC) Scale was created by Telch et al. (2004) to improve upon existing self-report measures of social anxiety-related cognition. In a largely nonclinical sample, the ASC was found to possess three factors and was psychometrically sound. In a smaller clinical sample, the ASC demonstrated sensitivity to the effects of cognitive behavioral therapy. In the present study, the psychometric properties of the ASC were examined in a larger sample of patients with social anxiety disorder. In this sample, the ASC exhibited a 2-factor structure; the nature of the factors was similar to the primary factors originally reported by Telch et al. The ASC also demonstrated strong validity, internal consistency, and sensitivity to treatment effects. It is concluded that the ASC may be useful in the assessment of cognition and cognitive change in patients with social anxiety disorder.

Adult↗