Comparison of real-time PCR and hemagglutination assay for quantitation of human polyomavirus JC.docVIP

Virology Journal BioMedCentral Short report Open Access Comparison of real-time PCR and hemagglutination assay for quantitation of human polyomavirus JC Moti L Chapagain, Taylor Nguyen, Thomas Bui, Saguna Verma and Vivek R Nerurkar* Address: Retrovirology Research Laboratory, Department of Tropical Medicine, Medical Microbiology and Pharmacology, Asia-Pacific Institute of Tropical Medicine and Infectious Diseases, John A. Burns of School of Medicine, University of Hawaii, 651 Ilalo Street, BSB 325AA, Honolulu, Hawaii 96813, USA Email: Moti L Chapagain - moti@; Taylor Nguyen - minh@; Thomas Bui - tuanb@; Saguna Verma - saguna@; Vivek R Nerurkar* - nerurkar@ * Corresponding author Published: 09 January 2006 Received: 04 September 2005 Accepted: 09 January 2006 Virology Journal 2006, 3:3 doi:10.1186/1743-422X-3-3 This article is available from: /content/3/1/3 ? 2006 Chapagain et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Human polyomavirus JC (JCV), the etiological agent of the disease progressive multifocal leukoencephalopathy (PML) affects immunocompromised patients particularly patients with AIDS. In vitro studies of JCV infection are hampered by the lack of sensitive JCV quantitation tests. Although the hemagglutination (HA) assay has been routinely employed for in vitro quantitation of JCV, its sensitivity is severely limited. We have employed a real-time PCR assay which compares favorably with the HA assay for the in vitro quantitation of JCV. JCV(Mad1), propagated in primary human fetal glial (PHFG) cells in two independent laboratories, was purified and quantitated by the HA assay. Both batches of purified JCV(Mad1) were then serially diluted in Dulbeccos M