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Clinical Chemistry 50: 1057-1060, 2004; 10.1373/clinchem.2003.030783
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(Clinical Chemistry. 2004;50:1057-1060.)
© 2004 American Association for Clinical Chemistry, Inc.

Technical Briefs

Universal Sensing Strategy for the Detection of Nucleic Acid Targets by Optical Biosensor Based on Surface Plasmon Resonance

Bi-feng Yuan, Yu-hua Hao and Zheng Tana

1 Laboratory of Biochemistry and Biophysics, College of Life Sciences, Wuhan University, Wuhan 430072, People’s Republic of China

aauthor for correspondence: fax 86-27-8788-2661, e-mail tanclswu@public.wh.hb.cn

The first 300 words of the full text of this article appear below.

PCR is an important technique for identifying specific nucleic acid targets; for example, sequences associated with diseases and pathogens in clinical, environmental, and food samples. Many techniques currently used for sequence-specific detection of PCR products either require manual processing or are limited in the speed or scale of analysis. The development of biosensors in recent years has provided promising techniques for efficient sequence-specific DNA analysis (1). Biosensors have been used to detect target sequences in PCR products. Most studies have been conducted with the commercially available biosensor BIAcore, which is based on surface plasmon resonance (SPR) technology. In those investigations, either sequence-specific oligonucleotide probe (2)(3)(4)(5)(6)(7)(8) or amplified products themselves (8)(9)(10)(11) were immobilized on the sensor chip, limiting their application to one sequence or to one particular sample. Such a strategy suffers several drawbacks: (a) it compromises the automation and high-throughput capability of such instruments when different targets are analyzed; (b) chip-to-chip variation makes it difficult to compare different measurements; and (c) probe immobilization and chip consumption make running costs substantial.

To overcome these drawbacks, we describe here a "one-chip-for-all" strategy that is capable of, in principle, detecting different target sequences by use of the same sensor chip. Target sequences are amplified by asymmetric PCR using a primer pair in which the low-concentration primer carries a common tag sequence that is identical to that of the oligonucleotide capture probe immobilized on the sensor chip. The PCR product is then injected and captured on the sensor chip, and its sequence identity can be further verified by use of a target-specific probe (see Fig. 1 in the Data Supplement that accompanies the online version of this Technical Brief at . . . [Full Text of this Article]






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Copyright © 2004 by the American Association for Clinical Chemistry.