Development of a real-time quantitative PCR assay for detection of a stable genomic region of BK virus

<p>Abstract</p> <p>Background</p> <p>BK virus infections can have clinically significant consequences in immunocompromised individuals. Detection and monitoring of active BK virus infections in certain situations is recommended and therefore PCR assays for detection of BK virus have been developed. The performance of current BK PCR detection assays is limited by the existence of viral polymorphisms, unknown at the time of assay development, resulting in inconsistent detection of BK virus. The objective of this study was to identify a stable region of the BK viral genome for detection by PCR that would be minimally affected by polymorphisms as more sequence data for BK virus becomes available.</p> <p>Results</p> <p>Employing a combination of techniques, including amino acid and DNA sequence alignment and interspecies analysis, a conserved, stable PCR target region of the BK viral genomic region was identified within the VP2 gene. A real-time quantitative PCR assay was then developed that is specific for BK virus, has an analytical sensitivity of 15 copies/reaction (450 copies/ml) and is highly reproducible (CV ≤ 5.0%).</p> <p>Conclusion</p> <p>Identifying stable PCR target regions when limited DNA sequence data is available may be possible by combining multiple analysis techniques to elucidate potential functional constraints on genomic regions. Applying this approach to the development of a real-time quantitative PCR assay for BK virus resulted in an accurate method with potential clinical applications and advantages over existing BK assays.</p

Renal replacement therapy in patients with acute respiratory distress syndrome: a single-center retrospective study

Joshua Dill,1 Billie Bixby,1 Huthayfa Ateeli,1 Benjamin Sarsah,2 Khushboo Goel,3 Ryan Buckley,3 Ilya Finkelshteyn,3 Bijin Thajudeen,2 Pradeep V Kadambi,4 Christian Bime1 1Division of Pulmonary, Allergy, Critical Care and Sleep Medicine, Department of Medicine, The University of Arizona, Tucson, AZ, USA; 2Division of Nephrology, Department of Medicine, The University of Arizona, Tucson, AZ, USA; 3General Internal Medicine, Department of Medicine, The University of Arizona, Tucson, AZ, USA; 4Department of Medicine, University of Florida College of Medicine &ndash; Jacksonville, Jacksonville, FL, USA Background: Patients with acute respiratory distress syndrome (ARDS) who develop acute kidney injury have increased mortality and frequently require renal replacement therapy (RRT). The optimal timing for initiation of RRT after onset of ARDS to improve survival is not known. Methods: We retrospectively reviewed clinical data on patients admitted to our health system over a 2-year period. Individual charts were carefully reviewed to ascertain that patients met the Berlin criteria for ARDS and to categorize RRT utilization. The Kaplan&ndash;Meier analysis was conducted to compare early (&pound;48&nbsp;hours postintubation) versus late (&gt;48&nbsp;hours postintubation) initiation of RRT. Associations between RRT initiation and mortality were evaluated using Cox proportional hazards regression. Results: A total of 75 patients were identified with ARDS, 95% of whom received RRT. Mortality of patients who required RRT was 56%. The main indications for RRT initiation were fluid overload (75%), metabolic acidosis (64%), and hyperkalemia (33%). The Kaplan&ndash;Meier analysis comparing early initiation of RRT to late initiation of RRT showed no survival benefit. Cox proportional hazard models testing the association between timing of RRT initiation with survival and adjusting for sex, race, ethnicity, and Acute Physiology and Chronic Health Evaluation II&nbsp;score did not reach statistical significance (HR=0.94, 95% CI=0.48&ndash;1.86). Conclusion: Timing of RRT initiation was not associated with a survival benefit. Prospective study in the utilization and outcomes of RRT in ARDS could assist in optimizing its usage in this population. Keywords: acute respiratory distress syndrome, AKI, dialysis, intensive care, rena

Complications of Pediatric Renal Transplantation

The major advance allowing prolonged graft survival has been the use of immunosuppressive drugs that downregulate the immune response. The immunosuppression that is used varies among centers and evolves with the development of new medications