Detection and Differentiation of Human Polyomaviruses JC and BK by LightCycler PCR

Abstract

Human polyomaviruses JC and BK may cause several clinical manifestations in immunocompromised hosts, including progressive multifocal leukoencephalopathy and hemorrhagic cystitis. Molecular detection by PCR is recognized as a sensitive and specific method for detecting human polyomaviruses in clinical samples. In this study, a real-time PCR assay using the LightCycler platform was evaluated and compared to an "in-house" PCR assay using a conventional detection method. A total of 122 urine specimens were tested, and human polyomavirus was detected in 49 specimens (40%) by both conventional PCR and LightCycler PCR. The remaining 73 specimens (60%) were found negative by both assays. For 46 of the 49 positive specimens, LightCycler PCR and conventional PCR identified the same polyomavirus type. These samples included 30 samples with JC virus (JCV), 14 samples with BK virus (BKV), and 2 samples in which both viruses were detected. In the remaining three samples, both JCV and BKV were detected by the conventional assay, but only JCV was detected by the LightCycler assay. The results of this study show that the LightCycler PCR assay displays sensitivity and specificity similar to those of a conventional PCR assay. These data, combined with its rapid turnaround time for results and decreased hands-on time, make the LightCycler PCR assay highly suitable for the rapid detection and differentiation of JCV and BKV in the clinical laboratory.

Detection and Differentiation of Mycobacterium tuberculosis and Nontuberculous Mycobacterial Isolates by Real-Time PCR

“…The ability of the LightCycler system to perform melting curve analyses also allows for differentiation of closely related organisms (24, 29; L. Doyle, C. Starkey, B. Yen-Lieberman, and G. W. Procop, presented at the 102nd General Meeting of the American Society for Microbiology, 2002). This feature has been used to distinguish between closely related organisms, such as the polyomaviruses JC virus and BK virus, Bartonella species, and Bordetella pertussis and Bordetella parapertussis (24,29; Doyle et al, presented at 102nd Gen. Meet. Am.…”

Urinary Cytokine Response In BK Virus Infection

“…So, PCR is now emerging as the "gold standard" for detection of polyomavirus (3). For this reason, in our study we determined the BKV infection in patients by using PCR methods.…”

“…Primary infections with BKV are usually asymptomatic and may result in transient viruria (3). After primary infection, BKV becomes latent in renal tissues and the peripheral blood (1).…”

Molecular Diagnosis of Medical Viruses

“…It has been particularly important in the rapid diagnosis of life-threatening viral diseases such as those caused by avian influenza (H5N1) (Whiley et al, 2005) or herpes simplex (herpes simplex encephalitis) (Whiley et al, 2004b). However, significant improvements have also been achieved in our laboratory in applying real-time PCR protocols for the detection of respiratory viruses, the human herpesviruses, enteroviruses, JC virus, BK virus, Bordetella pertussis, Mycoplasma pneumoniae, Chlamydia trachomatis, Neisseria gonorrhoea as well as Aspergillus fumigatus and Pneumocystis jiroveci (Whiley et al, 2004a;Whiley et al, 2001;Whiley et al, 2002). A comparison of these assays with conventional culturebased methods convincingly demonstrated greater sensitivity of the molecular assays.…”