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Clinical Chemistry 50: 1949-1952, 2004. First published August 19, 2004; 10.1373/clinchem.2004.036723
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(Clinical Chemistry. 2004;50:1949-1952.)
© 2004 American Association for Clinical Chemistry, Inc.

Abstracts of Oak Ridge Posters

Sensitive Assay for Identification of Methicillin-Resistant Staphylococcus aureus, Based on Direct Detection of Genomic DNA by Use of Gold Nanoparticle Probes

Ramesh Ramakrishnana, Wesley Buckingham, Marc Domanus, Linn Gieser, Karin Klein, Gregory Kunkel, Anna Prokhorova and Peter V. Riccelli

1 Nanosphere, Inc., Northbrook, IL 60062

aauthor for correspondence

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

Staphylococcus aureus (SA) is one of the most important human pathogens, causing both nosocomial and community-acquired infections (1)(2). The occurrence of methicillin-resistant SA (MRSA) has increased steadily worldwide and now accounts for a substantial portion of all staphylococcal infections in US hospitals (3). To develop preventive measures, a rapid screening method, along with accurate and timely identification of MRSA, is essential. The existing techniques for doing so are either time-intensive (culturing of bacteria on selective media), relatively insensitive (use of latex agglutination), or expensive and easily susceptible to operator error (such as PCR).

We describe a method designed for clinical laboratories using oligonucleotides conjugated to gold nanoparticles. We avoid radioactivity, fluorescence, or target amplification (such as PCR), and use a simple and rapid hybridization-based approach in a microarray format, with ClearReadTM technology to detect specific genomic sequences (4).

The ClearRead procedure involves a two-step process: the first involves the hybridization of target to oligonucleotides conjugated to gold nanoparticles as well as oligonucleotides attached to a solid matrix; the second step involves the catalytic deposition of silver on the gold nanoparticle, providing a sixfold amplification of signal (Fig. 1A ) (4)(5). The differentiation at isothermal hybridization temperatures is a result of the sharp melting transitions characteristic of nanoparticle probes (5). Previous methods based on this property (6)(7) have required the use of PCR and have not directly assayed for genomic DNA. Our assay, on the other hand, requires minimal amounts of genomic DNA (~500 ng, or ~108 DNA molecules) and has been used to reliably identify MRSA from liquid cultures, based on the detection of the mecA and tuf genes. Resistance to methicillin is mediated by the presence of penicillin-binding protein 2a, . . . [Full Text of this Article]




The following articles in journals at HighWire Press have cited this article:


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L.-X. Zhu, Z.-W. Zhang, C. Wang, H.-W. Yang, D. Jiang, Q. Zhang, K. Mitchelson, and J. Cheng
Use of a DNA Microarray for Simultaneous Detection of Antibiotic Resistance Genes among Staphylococcal Clinical Isolates
J. Clin. Microbiol., November 1, 2007; 45(11): 3514 - 3521.
[Abstract] [Full Text] [PDF]

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 Antimicrob. Agents Chemother.Home page
L.-X. Zhu, Z.-W. Zhang, D. Liang, D. Jiang, C. Wang, N. Du, Q. Zhang, K. Mitchelson, and J. Cheng
Multiplex Asymmetric PCR-Based Oligonucleotide Microarray for Detection of Drug Resistance Genes Containing Single Mutations in Enterobacteriaceae
Antimicrob. Agents Chemother., October 1, 2007; 51(10): 3707 - 3713.
[Abstract] [Full Text] [PDF]

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P. V. Baptista, M. Koziol-Montewka, J. Paluch-Oles, G. Doria, and R. Franco
Gold-Nanoparticle-Probe-Based Assay for Rapid and Direct Detection of Mycobacterium tuberculosis DNA in Clinical Samples.
Clin. Chem., July 1, 2006; 52(7): 1433 - 1434.
[Full Text] [PDF]


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