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PubMed2005

The cerebellar deficit hypothesis and dyslexic tendencies in a non-clinical sample.

In order to assess the relationship between cerebellar deficits and dyslexic tendencies in a non-clinical sample, 27 primary school children aged 8-9 completed a cerebellar soft signs battery and were additionally assessed for reading age, sequential memory, picture arrangement and knowledge of common sequences. An average measure of the soft signs data established a single construct which we treated as our primary index of cerebellar function. Overall cerebellar function was significantly correlated with reading age, picture arrangement and knowledge of common sequences even when IQ was partialled out. Graphical representation of our data indicated a continuous rather than discrete distribution for each measure. Results are discussed in terms of the CDH and continuum approaches to dyslexia.

Cerebellar Diseases↗
PubMed2003

Biotypic characterisation of bovine viral diarrhoea virus (BVDV) using reverse transcription-polymerase chain reaction (RT-PCR) in clinical samples.

In this study, bovine viral diarrhoea virus (BVDV) was detected and bio-typically characterised in clinical samples using reverse transcriptase-polymerase chain reaction (RT-PCR). The RT-PCR technique produced two different amplicons (402 and approx. 680 bp in size) in case of the presence of both biotypes (cp and ncp) in the sample. The mixture of the biotypes as detected by RT-PCR was verified by the immunoplaque assay (IPA). Purification of biotypes was carried out by native plaque isolation and subsequent RT-PCR revealed single products (402 or approx. 680 bp in size) in each clone. The results showed that RT-PCR can be used for accurate molecular differentiation between the BVDV biotypes.

Animals↗
PubMed2001

Quantification of lupus anticoagulants in clinical samples using anti-beta2GP1 and anti-prothrombin monoclonal antibodies.

Quantification of lupus anticoagulant (LA) in clinical samples is hampered by the lack of a suitable standard of activity. We evaluated the use of mAbs displaying LA activity for this purpose. As most patient samples contain both beta2Glycoprotein I (beta2GP1) and prothrombin dependent LA, a combination of two mAbs, one of each specificity, was added to normal plasma in a concentration from 0 to 60 microg/ml. Eight assay systems using different reagents and instruments were used. The calibration curves were linear for all but one, with marked differences between the responsiveness to each mAb. A panel of plasmas from 69 patients with persistent LA diagnosed using the SSC-ISTH criteria was tested. An antiphospholipid syndrome (APS) was present in 40, whereas 29 were asymptomatic. LA activities of individual plasmas varied between assays (p < 10(-4)), but homogeneous subgroups were identified. In a majority of samples, LA activity displayed a prothrombin-dependent profile, with a variable contribution of beta2GP1-dependent activity. The latter was associated to beta2GP1 antibodies detected by solid-phase immunoassay. By using 3 dilute Russell viper venom time assays, higher LA titers were found in APS, compared to asymptomatic patients (p <0.05).

Adult↗
PubMed2005

[Methods of extraction of Mycobacterium tuberculosis DNA from clinical samples for PCR: comparison and evaluation].

Several commonly used methods of extraction of Mycobacterium tuberculosis DNA from clinical samples for further use in PCR were compared. We used the commercial sets "DNA-express" (Litekh, Russia) and "DNA-sorb-B" (Amplisens, Russia) as well as phenol extraction, fast alkaline lysis and DNA sorption on silicone gel for DNA extraction. All methods were compared by the quantity of isolated DNA and stability of DNA preparation as well as by efficiency and cost of extraction. "DNA-express" proved to be most effective in the extraction of Mycobacterium tuberculosis DNA with its cost being the smallest.

DNA, Bacterial↗
PubMed1994

Lifetime DSM-IV diagnosis of alcohol, cannabis, cocaine and opiate dependence: six-month reliability in a multi-site clinical sample.

Psychiatric research increasingly emphasizes the diagnosis of symptoms and syndromes on a longitudinal basis. This study tests the reliability of lifetime DSM-IV diagnoses of alcohol, cannabis, cocaine and opiate dependence. The CIDI-SAM was administered at intervals not less than six months apart to a multi-site sample of 201 clinical respondents. The reliability of lifetime diagnosis of the syndromes, of the criteria which constitute the syndromes, and of the ages of onset reported for the criteria and for the dependence syndromes as a whole, were studied and the effects of patient characteristics suspected to degrade reliability were examined. There was generally good agreement, statistically, at both the syndrome and criterion level between the two interviews. Lifetime diagnoses for three of the drugs--alcohol, cannabis and opiates--were made at or near levels of agreement generally considered excellent under less strict testing conditions, and cocaine dependence was only marginally below this level. Most criteria showed good reliability and all delivered about equal results when averaged across the four substances, although a relationship between reliability and centrality of the symptom to the individual drug abuse pattern was found. Age of onset was almost uniformly highly reliable. Most patient characteristics bore no detectable relationship to reliability, although patients with multiple drug use patterns may warrant more careful probing by interviewers. Overall, these data indicate that lifetime symptoms and diagnoses can be queried reliably, although they must be reported with less confidence than current state diagnoses.

Age of Onset↗
PubMed1990

[Numerical taxonomy of Staphylococci isolated from clinical samples].

A taxonomic study of 82 strains of Staphylococci and 6 reference samples isolated from clinical cases was carried out. Results of 47 phenotype tests were submitted to a numerical analysis using the similarity coefficient of Sokal and Michener and a grouping technique (UPGMA). Based on a 75 similarity level, 4 phenons were identified. 37 strains identified as S aureus were grouped in phenon A; 32 strains of S hominis in phenon B; 5 strains of S epidermidis in phenon C and 8 strains of S ssp in phenon D.

Bacteriological Techniques↗
PubMed2002

Obsessive-compulsive disorder with and without tics in a clinical sample of children and adolescents.

The purpose of this study was to discriminate subtypes of obsessive-compulsive disorder (OCD) in a clinical sample of children and adolescents. Sixty OCD patients were assessed in two outpatient psychiatric clinics; 15 patients had a lifetime history of tics and 45 patients had no tic history. Interviews were conducted with the patients and their parents by a child psychiatrist using the Children's Yale-Brown Obsessive Compulsive Scale (CY-BOCS). The symptom checklist of the CY-BOCS was used to categorize obsessions and compulsions. Discriminant function analysis was used to compare the two groups in their symptomatology. There was no difference between the two groups in seven obsession categories. However, there was a significant difference between the two groups in seven compulsion categories. Ordering, hoarding, and washing compulsions were more common in those with no tic history. The results indicate that tic-related OCD may be differentiated from non-tic-related OCD early in life by the presence or absence of certain compulsive symptoms.

Adolescent↗
PubMed2005

[Anxiety disorders in an attention-deficit/hyperactivity disorder clinical sample].

OBJECTIVE: To evaluate the prevalence of anxiety disorders in a clinical referred sample of children and adolescents with attention deficit/hyperactivity disorder (ADHD). METHOD: 78 children and adolescents with ADHD according to DSM-IV criteria were investigated with a semi-structured interview (P-CHIPS), complemented by clinical interviews with the children or adolescents and their parents. Their IQ was calculated with neuropsychological testing. RESULTS: A high prevalence of anxiety disorders (23.05%) was found in the sample. Generalized anxiety disorder was the most prevalent disorder (12,8%), followed by social phobia (3,84%) and separation anxiety disorder (3,8%). Two children showed more than one anxiety disorder. CONCLUSION: Children and adolescents with ADHD seem to be more prone to have comorbid anxiety disorders, at least in clinical samples referred to specialized units.

Adolescent↗
PubMed1991

Polymerase chain reaction for detection of Mycoplasma genitalium in clinical samples.

We have used the polymerase chain reaction to detect Mycoplasma genitalium in artificially seeded human throat swab samples as well as in clinical material. On the basis of the published nucleotide sequence of the M. genitalium 140-kDa adhesin gene, primers were chosen to produce an amplified fragment of 281 bp. Five different previously isolated strains, including the type strain of M. genitalium, could all be detected by the polymerase chain reaction, and DNAs from other mycoplasmal and bacterial species yielded negative results. The detection limits were estimated to be approximately 50 organisms by inspection of ethidium bromide-stained agarose gels and 4 organisms when a biotinylated oligonucleotide probe was used in filter hybridization. The amplified DNA fragments were subjected to restriction enzyme analysis. DNAs from the five different isolates all possessed EcoRI, SspI, AluI, Sau3AI, and DdeI restriction sites, as predicted from the published sequence. A total of 150 urogenital swabs collected from 100 patients for culturing of Chlamydia trachomatis were tested for the presence of M. genitalium DNA. Ten samples from eight patients were found positive. The amplified DNA fragments from all of our clinical samples possessed the AluI, Sau3AI, and DdeI restriction sites, but three samples from two patients did not contain the SspI site and none of the samples contained the EcoRI site.

Base Sequence↗
PubMed2006

A novel fluorescence polarization based assay for 14 human papillomavirus genotypes in clinical samples.

A specific and practical method was developed for high throughput 14 human papillomavirus (HPV) genotypes assay in clinical samples by a single PCR. GP5+/6+ polymerase chain reaction (PCR) system was used to amplify HPV DNA in 1127 samples. The PCR product was assayed by AcycloPrime reaction with fluorescence polarization (FP). Fourteen HPV genotypes specific sequence primers designed within GP5+/GP6+ amplification polymorphism regions of L1 genes for corresponding HPV genotypes were annealed with the type specific PCR products and special fluorescent terminator was added to the end of the primer under direction of the PCR products. AcycloPrime-FP analysis showed specific anneal and incorporation without any cross-reaction. The types detected with FP showed an excellent overall agreement with sequence when the individual monotype results were taken into account. The proposed method could detect more than one type of HPV infection, but the sequence method was limited. AcycloPrime-FP could reach the detection level: 100 ag for representative phylogenetically distant HPV genotypes: HPV6, 18, 31, 39, 42, 51 and 58. The results of AcycloPrime-FP showed excellent reproducibility. The proposed method allowed an economical detection of HPV genotypes without any use of labeled probe. It is expected to be an extremely useful tool for HPV genotypes screening.

Capsid Proteins↗
PubMed1994

Stability of infectious HIV in clinical samples and isolation from small volumes of whole blood.

AIMS: To evaluate the stability of infectious HIV in clinical samples and the efficiency of isolating it from small volumes of whole blood. METHODS: Titres of infectious HIV were measured in peripheral blood mononuclear cells and plasma 2, 24, and 48 hours after peripheral blood had been collected from 10 HIV positive adult volunteers. Volumes of whole blood (1 ml to 5 microliters), collected from a further five volunteers, were used to determine the minimum volume from which HIV could be isolated. Infectious HIV was isolated by co-culture with phytohaemagglutinin stimulated umbilical cord mononuclear cells. RESULTS: Geometric mean titres of infectious HIV seemed to be more stable in peripheral blood mononuclear cells than in plasma. HIV was recovered from all 10 peripheral blood mononuclear cell samples during the 48 hours after sample collection, but from only four plasma samples. HIV could occasionally be isolated from 5 microliters of whole blood and reliably from 200 microliters. CONCLUSIONS: HIV can be isolated from peripheral blood mononuclear cells and plasma for up to 48 hours after sample collection. Isolation of HIV from small volumes of whole blood has applications for the diagnosis and management, of HIV positive children.

Adult↗
PubMed2005

[Comorbid eating disorders in a Brazilian attention-deficit/hyperactivity disorder adult clinical sample].

UNLABELLED: Although comorbidity between attention-deficit/hyperactivity disorder (ADHD) and eating disorders (ED) is relevant for clinical treatment, it is seldom investigated. METHODS: 86 DSM-IV attention deficity hyperactivity disorder patients out of 107 self-referred adults in a specialized center for attention deficity hyperactivity disorder were interviewed using SCID-R to evaluate the lifetime prevalence of ED and other comorbid conditions. RESULTS: Nine attention deficity hyperactivity disorder patients had comorbid eating disorders; binge eating disorder (BED) was the most common diagnosis. The group with eating disorders presented a higher prevalence of other comorbid disorders (p=0.02). No significant differences were found on gender, age at assessment, schooling level and type of attention deficity hyperactivity disorder between groups. CONCLUSION: Attention deficity hyperactivity disorder clinical samples may have a high prevalence of BED comorbidity. Patients with attention deficity hyperactivity disorder and eating disorders may have a different comorbid profile.

Adolescent↗
PubMed1999

Development of a high-throughput quantitative assay for detecting herpes simplex virus DNA in clinical samples.

We have developed a high-throughput, semiautomated, quantitative fluorescence-based PCR assay to detect and type herpes simplex virus (HSV) DNA in clinical samples. The detection assay, which uses primers to the type-common region of HSV glycoprotein B (gB), was linear from <10 to 10(8) copies of HSV DNA/20 microl of sample. Among duplicate samples in reproducibility runs, the assay showed less than 5% variability. We compared the fluorescence-based PCR assay with culture and gel-based liquid hybridization system with 335 genital tract specimens from HSV type 2 (HSV-2)-seropositive persons attending a research clinic and 380 consecutive cerebrospinal fluid (CSF) samples submitted to a diagnostic virology laboratory. Among the 162 culture-positive genital tract specimens, TaqMan PCR was positive for 157 (97%) specimens, whereas the quantitative-competitive PCR was positive for 144 (89%) specimens. Comparisons of the mean titer of HSV DNA detected by the two assays revealed that the mean titer detected by the gel-based system was slightly higher (median, 1 log). These differences in titers were in part related to the fivefold difference in the amount of HSV DNA used in the amplicon standards with the two assays. Among the 380 CSF samples, 42 were positive by both assays, 13 were positive only by the assay with the agarose gel, and 3 were positive only by the assay with the fluorescent probe. To define the subtype of HSV DNA detected in the screening assay, we also designed one set of primers which amplifies the gG regions of both types of HSV and probes which are specific to either HSV-1 (gG1) or HSV-2 (gG2). These probes were labeled with different fluorescent dyes (6-carboxyfluorescein for gG2 and 6-hexachlorofluorescein for gG1) to enable detection in a single PCR. In mixing experiments the probes discriminated the correct subtype in mixtures with up to a 7-log-higher concentration of the opposite subtype. The PCR typing results showed 100% concordance with the results obtained by assays with monoclonal antibodies against HSV-1 or HSV-2. Thus, while the real-time PCR is slightly less sensitive than the gel-based liquid hybridization system, the high throughput, the lack of contamination during processing, the better reproducibility, and the better ability to type the isolates rapidly make the real-time PCR a valuable tool for clinical investigation and diagnosis of HSV infection.

DNA, Viral↗
PubMed2005

Maternal mental representations of the child in an inner-city clinical sample: violence-related posttraumatic stress and reflective functioning.

Parental mental representations of the child have been described in the clinical literature as potentially useful risk-indicators for the intergenerational transmission of violent trauma. This study explored factors associated with the quality and content of maternal mental representations of her child and relationship with her child within an inner-city sample of referred, traumatized mothers. Specifically, it examined factors that have been hypothesized to support versus interfere with maternal self- and mutual-regulation of affect: posttraumatic stress disorder (PTSD) and maternal reflective functioning (RF). More severe PTSD, irrespective of level of RF, was significantly associated with the distorted classification of non-balanced mental representations on the Working Model of the Child Interview (WMCI) within this traumatized sample. Higher Levels of RF, irrespective of PTSD severity, were significantly associated with the balanced classification of maternal mental representations on the WMCI. Level of maternal reflective functioning and severity of PTSD were not significantly correlated in this sample. Clinical implications are discussed.

Adolescent↗
PubMed2004

Microarray-based identification of bacteria in clinical samples by solid-phase PCR amplification of 23S ribosomal DNA sequences.

The rapid identification of the bacteria in clinical samples is important for patient management and antimicrobial therapy. We describe a DNA microarray-based PCR approach for the quick detection and identification of bacteria from cervical swab specimens from mares. This on-chip PCR method combines the amplification of a variable region of bacterial 23S ribosomal DNA and the simultaneous sequence-specific detection on a solid phase. The solid phase contains bacterial species-specific primers covalently bound to a glass support. During the solid-phase amplification reaction the polymerase elongates perfectly matched primers and incorporates biotin-labeled nucleotides. The reaction products are visualized by streptavidin-cyanine 5 staining, followed by fluorescence scanning. This procedure successfully identified from pure cultures 22 bacteria that are common causes of abortion and sterility in mares. Using the on-chip PCR method, we also tested 21 cervical swab specimens from mares for the presence of pathogenic bacteria and compared the results with those of conventional bacteriological culture methods. Our method correctly identified the bacteria in 12 cervical swab samples, 8 of which contained more than one bacterial species. Due to the higher sensitivity of the on-chip PCR, this method identified bacteria in five cervical swab samples which were not detected by the conventional identification procedure. Our results show that this method will have great potential to be incorporated into the routine microbiology laboratory.

Abortion, Veterinary↗
PubMed1985

Comparison of WAIS and WAIS-R scaled scores for an outpatient clinic sample retested over extended intervals.

High correlations were obtained between WAIS and WAIS-R scores for an outpatient clinic sample tested and retested over extended periods. While the WAIS-R scores were generally lower than the WAIS values, the decrements were slight, suggesting that a previously administered WAIS can serve as a good estimate of what might be obtained from a second administration of the WAIS-R for this type of sample.

Adult↗
PubMed1996

A rapid, highly sensitive method for the detection of Francisella tularensis in clinical samples using the polymerase chain reaction.

We have developed a highly sensitive method for detection of Francisella tularensis in clinical samples based on a nested polymerase chain reaction (PCR) for the FopA gene. Mice infected with F. tularensis were killed at 24-hr intervals, and the DNA from blood and spleens was extracted by a variety of methods and analyzed by PCR. The best method, based on the ability of DNA to bind to silica in the presence of guanidine thiocyanate, yielded amplifiable DNA without dilution of the murine tissue samples. Francisella tularensis in infected murine spleens and culture-positive blood samples was reliably detected by nested PCR following this extraction procedure. We believe this technique has significant advantages over traditional methods for diagnosing F. tularensis infection in terms of speed, ease of use, reproducibility, and safety.

Animals↗
PubMed1986

Self-contained enzymic membrane immunoassay for detection of rotavirus antigen in clinical samples.

A self-contained enzymic membrane immunoassay (SCEMIA) system has been developed for the detection of viral antigens in clinical samples. The assay system makes use of antiviral antibodies bound to a nylon membrane, a flow-through washing procedure, and a clearly visible endpoint of the enzymic reaction. A SCEMIA system with antibodies against rotavirus detected rotavirus antigen, within 15 min, in all faecal samples from children with gastroenteritis that were positive for antigen in a standard microplate enzyme immunoassay, which took 4 h to complete. In addition, the SCEMIA could detect rotavirus in faecal samples collected from infected individuals both before and after antigen could be detected by a standard immunoassay system. Rotavirus antigen was not detectable in control children who did not have evidence of rotavirus infection. SCEMIA systems are an accurate, rapid, and inexpensive means for the practical diagnosis of viral infections in human beings.

Antigens, Viral↗