External quality assessment for the molecular detection of Hepatitis C virus.

UNLABELLED: BACKGROUND, OBJECTIVES AND STUDY DESIGN: External quality assessment (EQA) panels were distributed internationally by UK NEQAS for Microbiology to 159 participants for the detection, quantification and genotyping of Hepatitis C virus (HCV) in freeze-dried plasma from 2000 to 2004. The results were analysed to determine the level of standardisation of qualitative detection, quantitative detection and genotyping. RESULTS: The accurate detection of HCV in the panels varied from 86.9% to 100%. Four genotypes were distributed with the panels and there was no significant difference in the detection of different genotypes of HCV by participants. Further analysis indicated most variation occurred in quantification of HCV at lower concentrations and from 0% to 14.8% reported quantitative values outside 0.5 log(10) of the median value. In addition, three negative specimens were distributed and false positives were found to be rare (0.9-2.2%) with all methods included in the study. CONCLUSION: The laboratory detection of HCV in plasma EQA specimens was varied, with decreasing parity of quantification at lower concentrations of HCV. False positives and negatives were rare, irrespective of the genotype under test.

Examination of specimens for mycobacteria in clinical laboratories in 21 countries: a 10-year review of the UK National Quality Assessment Scheme for Mycobacteria Culture.

Results from clinical diagnostic microbiology laboratories taking part in the UK National Quality Assessment Service (UK NEQAS) scheme for Mycobacteria Culture between 1993 and 2003 were evaluated and assessed to determine whether the perceived increase in the use of rapid methods is improving time-to-positive reporting of results. Four simulated sputum specimens containing mycobacteria in mixed cultures with normal commensal organisms were distributed three times a year. Participating laboratories were required to report on the presence of 'mycobacteria' and on the time required to obtain a positive result. The overall level of performance with the mycobacteria culture external quality assessment specimens remained consistently high, with an average success rate of 94% over 10 years. The mean time-to-positive decreased from 24 to 17 days during the previous 8 years. A survey questionnaire, circulated in 2002, addressed the use of continuous automated mycobacterial liquid culture (CAMLiC) and molecular methods. The increase in the use of rapid culture methods for the detection of Mycobacterium tuberculosis has resulted in an overall reduction in time-to-positive data reported by participants, and has provided an indication of participants' ability to meet the 21-day target recommended by the CDC for the detection and identification of M. tuberculosis.

External quality assessment for detection of Chlamydia trachomatis.

The use of molecular methods for detection of Chlamydia trachomatis is increasing in clinical laboratories. External quality assessment enables unbiased monitoring of the performance of laboratories in the detection of specific pathogens. This study details the results of molecular and enzyme immunosorbent assay (EIA) testing for C. trachomatis detection in simulated endocervical swab specimens recently distributed internationally by United Kingdom National External Quality Assessment Scheme for Microbiology (UK NEQAS for Microbiology) external quality assessment panels. The frequency of accurate detection of C. trachomatis in the panels ranged from 32 to 100%. Participants using molecular methods were significantly more likely to detect C. trachomatis in specimens than those using an EIA. Two strains were distributed with the panels: an L2 laboratory-adapted strain and an uncharacterized primary isolate. Further analysis indicated a difference in detection of C. trachomatis between specific methods only with the L2 strain at lower concentrations. In addition, eight negative specimens were distributed, and false positives were found to be rare by all methods included in the study.

Seroepidemiology of human group C rotavirus in South Africa.

Sera from three separate healthy population cohorts were used to determine the incidence of group C rotavirus infections in 1,356 South Africans. Using an enzyme-linked immunosorbent assay based on a recombinant group C rotavirus VP6 protein, the total percent positivity was found to be 34.4% (range, 33 to 38%), with almost half of the population infected after the age of 20 years.

Infection with group C rotavirus in a suburban community in Brazil.

Group C rotaviruses are associated with sporadic outbreaks of gastroenteritis worldwide. Age-specific seroprevalence of group C rotavirus antibodies was investigated in sera, randomly collected and representative of a suburban community in Brazil which had previously been screened for group A rotavirus antibodies. Antibody prevalence to group C rotavirus was low in children under 5 years and increased slowly with age to 36% seropositivity in adults, reflecting continuous exposure to primary infection in all age groups. This suggests a higher incidence of infection than disease might predict. Adult antibody prevalence was similar to that in other geographical settings. No obvious patterns of infection with group A and group C rotavirus were found within individuals, which suggests independent transmission. However, further epidemiological studies are required to understand group C rotavirus dynamics and possible interactions with group A rotavirus transmission and immunity.

Enzyme-linked immunosorbent assay based on recombinant human group C rotavirus inner capsid protein (VP6) To detect human group C rotaviruses in fecal samples.

A recent study showed that 43% of a population in the United Kingdom were seropositive for group C rotavirus. The higher than expected incidence may be due to limited diagnosis of acute human group C rotavirus infections because no routine test is available. Human group C rotavirus infections are routinely diagnosed by electron microscopy (EM) and a negative group A rotavirus enzyme-linked immunosorbent assay (ELISA) result. An antigen-detection ELISA was developed with hyperimmune antibodies raised to human group C rotavirus recombinant VP6 (Bristol strain) expressed in insect cells. The assay was used to screen fecal samples to determine the prevalence of group C rotavirus infection. Samples positive by ELISA were confirmed by EM, polyacrylamide gel electrophoresis of double-stranded RNA, or detection of the VP6 gene by reverse transcription-PCR. Retrospective analysis indicated a 1 to 2% detection rate of positivity among samples from patients with acute diarrhea.

Seroepidemiology of human group C rotavirus in the UK.

The gene coding for the major inner capsid protein VP6 of human group C rotavirus was cloned into baculovirus using the pBlueBac2 vector and expressed in insect cells. When cultured in High Five cells, VP6 was expressed at a high level and exported to the cell culture medium. Purified VP6 was used to immunise rabbits. Hyperimmune rabbit serum, which reacted with native human group C rotavirus in infected cells, was used to develop and optimise an EIA for the detection of antibodies to group C rotavirus using the recombinant VP6 as a source of antigen. In a local epidemiological survey of 1000 sera grouped by age, an average of 43% of samples were found to have antibodies to human group C rotavirus with the highest proportion (66%) in the 71-75 year age group. In comparison, 97% of adults and 85% of children had antibodies to recombinant VP6 from the bovine RF strain of group A rotavirus. These results suggest that infection with human group C rotavirus is a common occurrence despite the apparent rarity of reports of human group C rotavirus in clinical samples from patients with gastroenteritis.

Anti-neurofilament protein antibodies in opsoclonus-myoclonus.

Opsoclonus-myoclonus (OM) is a neurological disorder usually occurring in infancy, clinically manifested by various involuntary movements. The pathogenesis of OM is unknown, but since the disease often is associated with viral infection or with neuroblastoma, an immunologic basis for OM has been postulated. We have studied two children with OM whose serum contained antibodies directed against the 210 kDa neurofilament protein; these antibodies were not seen in the serum of 21 children with other neurological disorders. Neurofilament proteins, which are found only in neurons, may be of prime importance in neuronal function, especially during development of the nervous system. Our findings suggest that generation of antibodies to the neurofilament proteins can occur in patients with opsoclonus-myoclonus; the role of the anti-NF210K antibodies in the pathogenesis of OM, however, is uncertain.

The use of picture cues to establish self-control in the preparation of complex meals by mentally retarded adults.

This investigation examined the effectiveness of picture cues in establishing self-control in the completion of complex meals by mildly and moderately retarded adults. Three participants, who lived in nonsheltered residential settings, were trained to prepare five complex meals. Following a training baseline comprised of pre-instruction, instructional feedback, and trainers' presence, picture recipe cards were introduced in a multiple-baseline fashion. Rapid improvement in the ability of each participant to independently complete each meal when the picture recipe cards were used occurred. A return to baseline for one of the participants demonstrated further self-directed antecedent stimulus control of the picture recipe cards as an effective treatment procedure. A discussion of future areas of research is included.