Rational Design of Functional Peptide Nucleic Acids for the Direct and Ultra-High Sensitive Detection of HIV-1 DNA and RNA with a Sandwich-Hybridization Assay

Thursday, October 11, 2012 — Poster Session III
10:00 a.m. – Noon	Natcher Conference Center, Building 45	NIDDK	TECH-13

Authors

• C. Zhao
• C. M. Micklitsch
• B. Y. Oquare
• D. H. Appella

Abstract

One of the great challenges in human immunodeficiency virus (HIV) diagnosis and prevention today is to develop technologies for early and direct detection of HIV nucleic acid sequences in patient tissue or blood samples. Here, a convenient, universal, colorimetric, nucleic acid-responsive detection system that uses two short peptide nucleic acids (PNAs) is demonstrated for the detection of HIV-1 gag DNA and RNA on a 96-well plate based on the sandwich-hybridization strategy. This protocol eliminates the requirement for a PCR step, and greatly improves the detection devices by using PNA probes instead of traditional DNAs for its outstanding properties. Furthermore, the design of a 3-cyclopentane modified surface probe and a 16-biotin containing reporter probe impart extraordinarily high sensitivity. This sandwich-hybridization assay is convenient, universal and colorimetric with a qualitative detection limit of 6 molecules for both of HIV-1 gag DNA and RNA, and a quantitative detection limit of 576 molecules for HIV-1 gag DNA, 493 molecules for HIV-1 gag RNA. In principle, this assay can also be used to detect any kind of infectious disease by simply changing the PNA sequences of the specific probe.

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