title: Evaluation of different urine protocols and DNA extraction methods for quantitative detection of BK viruria in kidney transplant patients.
abstract: Most transplant centers screen kidney transplant recipients for BK virus (BKV) infection using molecular techniques for the virus load determination. However, there is no consensus about the pre-analytical methods involved in the viral detection. In this study BK viral load was compared by the means of two urine treatment protocols (pelleted vs. whole urine) and two commercial DNA extraction kits for a quantitative PCR (qPCR) experiment. Ten patients who presented decoy cells in their urine sediment were selected for the study. Viral load was considerable higher (>1.5 log) for pelleted urine, in comparison to whole urine but no significant difference was observed between the extraction kits. PCR inhibition did not occur by using pelleted urine. In order to increase test sensitivity to detect BK viruria, pelleted urine should be the preferred urine compartment for qPCR experiments.
authors.affiliation: Universidade Federal de Ciências da Saúde de Porto Alegre-UFCSPA, Porto Alegre, Brazil.
authors.forename: Gabriel Godinho
authors.initials: GG
authors.lastname: Pinto
authors.forename: José Antonio Tesser
authors.initials: JA
authors.lastname: Poloni
authors.forename: Lilian Carla
authors.initials: LC
authors.lastname: Carneiro
authors.forename: Ludmila Fiorenzano
authors.initials: LF
authors.lastname: Baethgen
authors.forename: Afonso Luís
authors.initials: AL
authors.lastname: Barth
authors.forename: Alessandro Comarú
authors.initials: AC
authors.lastname: Pasqualotto
mesh_terms: D001739:BK Virus / Q000235:genetics / Q000302:isolation & purification*; D004279:DNA, Viral / Q000302:isolation & purification*; D006801:Humans; D016867:Immunocompromised Host; D016030:Kidney Transplantation / Q000009:adverse effects; D027601:Polyomavirus Infections / Q000175:diagnosis* / Q000821:virology; D013048:Specimen Handling / Q000379:methods*; D014556:Urine / Q000821:virology*; D019562:Viral Load / Q000379:methods*
chemical_list: D004279:DNA, Viral