A previous study suggested that delayed sample preparation can cause a false-positive plasma cytomegalovirus (CMV) PCR. We evaluated 78 samples submitted for CMV testing, and we report that a 24-h delay in plasma separation did not affect viral quantitation by real-time PCR. Laboratories should evaluate specimen handling requirements for their own PCR technology.Cytomegalovirus (CMV) is an important cause of morbidity and mortality in immunocompromised individuals (2). Accordingly, monitoring of patients for CMV viremia has become an important laboratory tool for transplantation services. We recently developed a real-time PCR assay for detection of CMV. Studies comparing our CMV PCR assay with the older CMV antigenemia assay (fluorescent antibody stain for expression of pp65 on mononuclear cells) have demonstrated that PCR is superior for early detection of CMV-related disease (1). After discharge from our institution, posttransplant patients are monitored for CMV viremia for up to 1 year. Because this often requires shipping of samples from distant sites, it is critical to validate specimen handling strategies to ensure correct results.For in-house patients, our PCR specimen handling protocols call for plasma to be separated immediately from EDTA-anticoagulated whole blood and stored frozen prior to testing. In contrast, the antigenemia test previously used for outpatients utilized EDTA-anticoagulated whole blood that was shipped and stored at room temperature for up to 48 h before processing. Although room temperature shipping of EDTA-anticoagulated whole blood is logistically simpler than immediate separation of plasma, it is unclear whether such handling allows accurate quantitation of CMV by real-time PCR. Shafer et al. reported that delayed separation of plasma increased the frequency of positive PCR results for CMV in latently infected patients (3). Those authors suggested that lysis of latently infected leukocytes released CMV into the plasma and thus increased the number of positive results. Shafer et al. used semiquantitative nested PCR followed by hybridization to radiolabeled probe, gel electrophoresis, and autoradiographic detection of amplified product. In contrast, our laboratory has used a rapid real-time TaqMan assay performed on the ABI 7700 (Boeckh et al., submitted). Schafer's assay amplified a region within the coding sequence of glycoprotein B (gB), while our amplicon spanned the junction between gB and UL123. Because of these differences, we felt it necessary to investigate whether delayed separation of plasma from EDTAanticoagulated blood would increase the frequency of detection of CMV DNA by our assay.To evaluate the potential release of CMV DNA from cells in whole blood, it was necessary to establish the stability of CMV DNA in plasma after separation from cells. Ten previously PCR-positive plasma samples were chosen to encompass our clinical measuring range. These samples, stored at Ϫ20°C, were thawed, and a portion was separated for extraction and CMV DNA quantity determination. The ...

Abstr Abstract actWhile recording traditionally has been viewed as a practical, adjunctive role of the music therapist, here the authors examine the skillful use of recording devices and software as fertile ground for the development of therapeutic programs with tangible benefits for adult clients in a concurrent disorders recovery setting. The integration and layering of musical composition with musical performance, digital technologies, and production invite rich and engaging conversations about therapeutic goals, processes, and outcomes. Using program evaluation and reflections on practice, the authors discuss how their interactions with clients through recording have yielded new insights into therapist roles and identities as well as expressions of music therapy. We outline the case for therapy-oriented recording, and a description of the authors' setting and information collection methods identified before a literature review on the use of recording in music therapy. The authors then distinguish four types of therapeutic recording illustrated by case examples from work with clients. The article culminates with a discussion of challenges and benefits associated with therapeutic recording. The authors conclude that recording offers critical and rewarding, yet often unrecognized, opportunities for music therapists to be innovators in their field.

Quantitative DNA detection of cytomegalovirus (CMV) and BK virus (BKV) is critical in the management of transplant patients. Quantitative laboratory-developed procedures for CMV and BKV have been described in which much of the processing is automated, resulting in rapid, reproducible, and high-throughput testing of transplant patients. To increase the efficiency of such assays, the performance and stability of four commercial preassembled frozen fast qPCR master mixes (Roche FastStart Universal Probe Master Mix with Rox, Bio-Rad SsoFast Probes Supermix with Rox, Life Technologies TaqMan FastAdvanced Master Mix, and Life Technologies Fast Universal PCR Master Mix), in combination with in-house designed primers and probes, was evaluated using controls and standards from standard CMV and BK assays. A subsequent parallel evaluation using patient samples was performed comparing the performance of freshly prepared assay mixes versus aliquoted frozen master mixes made with two of the fast qPCR mixes (Life Technologies TaqMan FastAdvanced Master Mix, and Bio-Rad SsoFast Probes Supermix with Rox), chosen based on their performance and compatibility with existing PCR cycling conditions. The data demonstrate that the frozen master mixes retain excellent performance over a period of at least 10 weeks. During the parallel testing using clinical specimens, no difference in quantitative results was observed between the preassembled frozen master mixes and freshly prepared master mixes. Preassembled fast real-time qPCR frozen master mixes perform well and represent an additional strategy laboratories can implement to reduce assay preparation times, and to minimize technical errors and effort necessary to perform clinical PCR.