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Journal of Clinical Microbiology, May 2007, p. 1581-1587, Vol. 45, No. 5
0095-1137/07/$08.00+0     doi:10.1128/JCM.01024-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Development of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of BK Virus{triangledown}

Bipin Raj Bista,1 Chandra Ishwad,1 Robert M. Wadowsky,2 Pradip Manna,3 Parmjeet Singh Randhawa,4 Gaurav Gupta,1 Meena Adhikari,5 Rakhi Tyagi,5 Gina Gasper,1 and Abhay Vats1*

Department of Pediatrics, Children's Hospital of Pittsburgh, 3705 Fifth Avenue, Pittsburgh, Pennsylvania 15213,1 Department of Pathology, Children's Hospital of Pittsburgh, 3705 Fifth Avenue, Pittsburgh, Pennsylvania 15213,2 Department of Molecular Diagnostics, ViraCor Laboratories, 1210 NE Windsor Drive, Lee's Summit, Missouri 64086,3 Department of Pathology, University of Pittsburgh Medical Center, 200 Lothrop Street, Pittsburgh, Pennsylvania 15213,4 Department of Molecular Virology, Atharva BioSciences, New Delhi 110029, India5

Received 16 May 2006/ Returned for modification 6 July 2006/ Accepted 14 December 2006

Loop-mediated isothermal amplification (LAMP) is a novel method for rapid amplification of DNA. Its advantages include rapidity and minimal equipment requirement. The LAMP assay was developed for BK virus (BKV), which is a leading cause of morbidity in renal transplant recipients. The characteristics of the assay, including its specificity and sensitivity, were evaluated. BKV LAMP was performed using various incubation times with a variety of specimens, including unprocessed urine and plasma samples. A ladder pattern on gel electrophoresis, typical of successful LAMP reactions, was observed specifically only for BKV and not for other viruses. The sensitivity of the assay with 1 h of incubation was 100 copies/tube of a cloned BKV fragment. Additionally, a positive reaction was visually ascertained by a simple color reaction using SYBR green dye. BKV LAMP was also successful for urine and plasma specimens without the need for DNA extraction. Due to its simplicity and specificity, the LAMP assay can potentially be developed for "point of care" screening of BKV.

* Corresponding author. Mailing address: Department of Pediatrics, Children's Hospital of Pittsburgh, 3705 Fifth Avenue, Pittsburgh, PA 15213. Phone: (412) 692-5182. Fax: (412) 692-7443. E-mail: abhay.vats{at}chp.edu

{triangledown} Published ahead of print on 21 February 2007.

Journal of Clinical Microbiology, May 2007, p. 1581-1587, Vol. 45, No. 5
0095-1137/07/$08.00+0     doi:10.1128/JCM.01024-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Hill, J., Beriwal, S., Chandra, I., Paul, V. K., Kapil, A., Singh, T., Wadowsky, R. M., Singh, V., Goyal, A., Jahnukainen, T., Johnson, J. R., Tarr, P. I., Vats, A. (2008). Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Common Strains of Escherichia coli. J. Clin. Microbiol. 46: 2800-2804 [Abstract] [Full Text]  


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