Skip to Content
Merck
  • Quantification of plasma HIV RNA using chemically engineered peptide nucleic acids.

Quantification of plasma HIV RNA using chemically engineered peptide nucleic acids.

Nature communications (2014-10-07)
Chao Zhao, Travis Hoppe, Mohan Kumar Haleyur Giri Setty, Danielle Murray, Tae-Wook Chun, Indira Hewlett, Daniel H Appella
ABSTRACT

The remarkable stability of peptide nucleic acids (PNAs) towards enzymatic degradation makes this class of molecules ideal to develop as part of a diagnostic device. Here we report the development of chemically engineered PNAs for the quantitative detection of HIV RNA at clinically relevant levels that are competitive with current PCR-based assays. Using a sandwich hybridization approach, chemical groups were systematically introduced into a surface PNA probe and a reporter PNA probe to achieve quantitative detection for HIV RNA as low as 20 copies per millilitre of plasma. For the surface PNA probe, four cyclopentane groups were incorporated to promote stronger binding to the target HIV RNA compared with PNA without the cyclopentanes. For the reporter PNA probe, 25 biotin groups were attached to promote strong signal amplification after binding to the target HIV RNA. These general approaches to engineer PNA probes may be used to detect other RNA target sequences.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Acetonitrile solution, contains 0.1 % (v/v) trifluoroacetic acid, suitable for HPLC
Supelco
Sulfuric acid, for the determination of nitrogen, ≥97.0%
Sigma-Aldrich
Diethyl ether
Sigma-Aldrich
Piperidine, ReagentPlus®, 99%
Sigma-Aldrich
N,N-Dimethylformamide, anhydrous, 99.8%
Sigma-Aldrich
N-Ethyldiisopropylamine, ≥98.0%
Supelco
Acetonitrile(Neat), Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Sulfuric acid, 99.999%
Sigma-Aldrich
Sulfuric acid, puriss. p.a., for determination of Hg, ACS reagent, reag. ISO, reag. Ph. Eur., 95.0-97.0%
Sigma-Aldrich
Sulfuric acid, ACS reagent, 95.0-98.0%
Sigma-Aldrich
Sulfuric acid solution, puriss. p.a., ≥25% (T)
Sigma-Aldrich
Sulfuric acid, puriss., meets analytical specification of Ph. Eur., BP, 95-97%
Sigma-Aldrich
Sodium carbonate, anhydrous, free-flowing, Redi-Dri, ACS reagent, ≥99.5%
Sigma-Aldrich
Sodium carbonate, BioUltra, anhydrous, ≥99.5% (calc. on dry substance, T)
Supelco
Sodium carbonate, reference material for titrimetry, certified by BAM, >99.5%
Sigma-Aldrich
Sodium carbonate, anhydrous, powder or granules, free-flowing, Redi-Dri, ACS reagent, ≥99.5%
Sigma-Aldrich
Sodium carbonate, BioXtra, ≥99.0%
Sigma-Aldrich
Sodium carbonate, anhydrous, powder, 99.999% trace metals basis
Sigma-Aldrich
Sodium carbonate, ReagentPlus®, ≥99.5%
Sigma-Aldrich
Sodium carbonate, ACS reagent (primary standard), anhydrous, 99.95-100.05% dry basis
Sigma-Aldrich
Sodium carbonate, powder, ≥99.5%, ACS reagent
Sigma-Aldrich
Sodium carbonate, ACS reagent, anhydrous, ≥99.5%, powder or granules
Sigma-Aldrich
Diethyl ether, ACS reagent, anhydrous, ≥99.0%, contains BHT as inhibitor
Sigma-Aldrich
Diethyl ether, suitable for HPLC, ≥99.9%, inhibitor-free
Sigma-Aldrich
Diethyl ether, contains 1 ppm BHT as inhibitor, anhydrous, ≥99.7%
Sigma-Aldrich
Diethyl ether, anhydrous, ACS reagent, ≥99.0%, contains BHT as inhibitor
Sigma-Aldrich
Trifluoromethanesulfonic acid, reagent grade, 98%
Sigma-Aldrich
Trifluoromethanesulfonic acid, ReagentPlus®, ≥99%
Sigma-Aldrich
Thioanisole, ReagentPlus®, ≥99%
Sigma-Aldrich
N,N-Dimethylformamide, ACS reagent, ≥99.8%