Development and validation with clinical samples of internally controlled multiplex real-time PCR for diagnosis of BKV and JCV infection in associated pathologies

• Published in: Comparative immunology, microbiology and infectious diseases microbiology and infectious diseases, 2012-03, Vol.35 (2), p.173-179
• Main Authors: Bárcena-Panero, Ana, Echevarría, Juan E, Romero-Gómez, María Pilar, Royuela, Enrique, Castellanos, Ana, González, Irene, Fedele, Giovanni
• Format: Article
• Language: English
• Subjects: Acquired immune deficiency syndrome; Allergy and Immunology; Biological and medical sciences; BK Virus - genetics; BK Virus - isolation & purification; Blood; Cerebrospinal fluid; Diagnosis; DNA, Viral - chemistry; Fundamental and applied biological sciences. Psychology; Human immunodeficiency virus; Humans; Infection; Infectious Disease; Internal control; JC Virus - genetics; JC Virus - isolation & purification; Kidney transplantation; Microbiology; Multiplex Polymerase Chain Reaction - methods; Nephropathy; Polymerase chain reaction; Polyomavirus Infections - diagnosis; Polyomaviruses; Progressive multifocal leukoencephalopathy; Quality Control; Quantification; Real-time PCR; Real-Time Polymerase Chain Reaction - methods; Reference Standards; Reproducibility of Results; Sensitivity and Specificity; Transplants; Tumor Virus Infections - diagnosis; Urine
• ISSN: 0147-9571; 1878-1667
• DOI: 10.1016/j.cimid.2011.12.010

Abstract This article describes the development and validation with clinical samples of an internally controlled multiplex quantitative real-time PCR (QRT-PCR) for human polyomaviruses BK (BKV) and JC (JCV). Blood and urine samples from renal transplant recipients with suspected nephropathy, and cerebrospinal fluid (CSF) specimens from AIDS, natalizumab-treated and HIV-negative patients with suspected progressive multifocal leukoencephalopathy, previously checked for BKV and JCV by conventional PCR, were tested by QRT-PCR. All samples positive by conventional PCR were confirmed by QRT-PCR. Four cases of JCV-associated neurological infection, including all those detected in natalizumab-treated patients, and one case of BKV-related neurological infection were only identified by QRT-PCR. BKV was quantified in the CSF of neurological patients for the first time. Analyses of the Quality Control for Molecular Diagnostics 2010 panel were “highly satisfactory” for BKV and “satisfactory” for JCV. The QRT-PCR is specific and reproducible. It improves the sensitivity of conventional PCR for the diagnosis of BKV and JCV infection in various diseases.