This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Dineva, M. A. Articles by Lee, H. Search for Related Content PubMed PubMed Citation Articles by Dineva, M. A. Articles by Lee, H.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, August 2005, p. 4015-4021, Vol. 43, No. 8
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.8.4015-4021.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Simultaneous Visual Detection of Multiple Viral Amplicons by Dipstick Assay

Department of Haematology, University of Cambridge, Cambridge CB2 2PT, United Kingdom,¹ National Blood Service Cambridge, Cambridge CB2 2PT, United Kingdom²

Received 15 November 2004/ Returned for modification 30 March 2005/ Accepted 6 May 2005

A sensitive, simple, and instrument-independent method for the visual detection and identification of multiple nucleic acid amplicons by dipstick has been developed. This method is based on nucleic acid hybridization on the dipstick membrane and a signal amplification system to allow visual detection. With hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus type 1 (HIV-1) as model analytes, it is demonstrated that the visual dipstick test combined with multiplex reverse transcription (RT)-PCR for the amplification of viral nucleic acid provides a specific and sensitive detection method. The RT-PCR products were detected by the dipstick with an efficiency similar to that of a complex, expensive, and instrument-dependent method based on fluorogenic oligonucleotide probes. The detection limits of the dipstick combined with multiplex RT-PCR were 50, 125, and 500 IU/ml for HBV DNA, HCV RNA, and HIV-1 RNA, respectively. The dipstick assay detected with similar efficiencies amplicons derived from strains of HBV genotypes A through F, HCV genotypes 1 to 6, and HIV-1 subtypes A through H as well as CRF02 circulating recombinant forms of HIV-1. Analysis of 295 clinical samples and 19 pools of 10 plasma specimens from blood donors revealed that multiplex dipstick detection was reproducible, sensitive, and specific. The visual dipstick detection of multiple amplicons thus provides an attractive alternative to complex, instrument-dependent detection methods currently in use for nucleic acid testing. This new and sensitive method for nucleic acid detection should increase the availability of genomic screening in resource-limited settings and its applicability to near-patient testing.

This article has been cited by other articles:

• Pilcher, C. D., Hare, C. B. (2008). The Deadliest Catch: Fishing for HIV in New Waters. ANN INTERN MED 149: 204-205 [Full Text]  
• Carter, D. J., Cary, R. B. (2007). Lateral flow microarrays: a novel platform for rapid nucleic acid detection based on miniaturized lateral flow chromatography. Nucleic Acids Res 35: e74-e74 [Abstract] [Full Text]  
• Panhotra, B. R., Hassan, Z. U., Joshi, C. S., Bahrani, A., Dineva, M. A., Allain, J.-P., Lee, H. (2005). Visual Detection of Multiple Viral Amplicons by Dipstick Assay: Its Application in Screening of Blood Donors a Welcome Tool for the Limited Resource Settings. J. Clin. Microbiol. 43: 6218-6219 [Full Text]