Rapid quantitation of cytomegalovirus DNA in whole blood by a new molecular assay based on automated sample preparation and real-time PCR.

Quantitation of cytomegalovirus (CMV) DNA in whole blood samples has gained significance in the routine diagnostic laboratory. In this study, the analytical performance of the artus™ CMV RG PCR kit in conjunction with automated sample preparation on the new QIAsymphony™ SP instrument was evaluated. Clinically referred samples were tested and results compared to those obtained with the routinely used molecular test system. Accuracy testing showed results to be within ±0.2 log(10) unit of the expected panel results. The assay was linear (R = 0.9972) from a lower quantification limit of 148 copies/ml to 1.3 × 10(7) copies/ml. The between-day imprecision CV was 8-63%, and the within-run imprecision CV was 16-61%. When 100 clinically referred samples were tested, results obtained with the new test system showed an acceptable concordance with those obtained with the routinely used easyMAG™ sample preparation and CMV HHV6,7,8 R-gene™ test system. Discrepant results were found with low-titer samples containing CMV DNA concentrations under the lower limit of quantification or within half a log unit above. The time to results was comparable for both test systems. The QIAsymphony™ sample preparation and artus™ CMV RG PCR test system allows for a rapid, sensitive, precise, and accurate high-throughput quantitation of CMV DNA in whole blood in the routine diagnostic laboratory.

Detection and quantitation of Epstein-Barr virus (EBV) DNA in EDTA whole blood samples using automated sample preparation and real time PCR.

BACKGROUND: Detection and quantitation of Epstein-Barr virus (EBV) DNA in EDTA whole blood samples has gained significance in the routine diagnostic laboratory. METHODS: In this study, the analytical and clinical performance of the artus EBV RG PCR kit in conjunction with automated sample preparation on the QIAsymphony SP instrument was evaluated. RESULTS: When the accuracy of the new test system was tested, all results were found to be within +/-0.5 log(10) unit of the expected panel results. Determination of linearity showed a quasilinear curve over 4 log units. The lower limit of detection was determined to be 391 EBV DNA copies/mL in EDTA whole blood. The between day imprecision ranged from 18% to 66%, and the within run imprecision ranged from 11% to 50%. When clinical samples were tested and the results compared with those obtained with the routinely used easyMAG sample preparation and EBV R-gene test system, 60 samples tested positive and 31 samples tested negative by both assays. Nineteen samples were found to be positive using the QIAsymphony sample preparation and artus EBV RG PCR test system only, and no samples tested positive with the routinely used test system only. CONCLUSIONS: The QIAsymphony sample preparation and artus EBV RG PCR test system is suitable for the detection and quantitation of EBV DNA in EDTA whole blood in the routine diagnostic laboratory.

Teachers' survey on problems with handwriting: referral, evaluation, and outcomes.

OBJECTIVE: The purpose of this study was to describe the factors that led elementary school teachers to refer students with handwriting difficulties to occupational therapy, the criteria they used to determine acceptable handwriting, and the handwriting outcomes they looked for after occupational therapy services. METHOD: A paper questionnaire composed of 31 close-ended questions was mailed to 400 first- through fourth-grade regular education teachers from 32 states to collect data from the 2000-2001 school year. The data were analyzed descriptively using frequency counts and converted to percentages. RESULTS: Information was obtained from 314 teachers. The main factor for handwriting referral to occupational therapy was that the student was not improving with classroom assistance alone. Teachers chose not being able to read student's writing as the main criterion they used to determine if the student's handwriting was acceptable, and increased legibility was the most important outcome they desired following occupational therapy services for handwriting remediation. CONCLUSION: The perceptions of regular education teachers on problems with handwriting can provide valuable information to occupational therapy practitioners when providing consultation and direct services related to handwriting in schools.