Engineering insights for multiplexed real-time nucleic acid sequence-based amplification (NASBA): implications for design of point-of-care diagnostics.

ABSTRACT:

Background

Nucleic acid sequence-based amplification (NASBA) offers huge potential for low-cost, point-of-care (POC) diagnostic devices, but has been limited by high false-positive rates and the challenges of primer design.

Objective

We offer a systematic analysis of NASBA design with a view toward expanding its applicability.

Methods

We examine the parameters that effect dimer formations, and we provide a framework for designing NASBA primers that will reduce false-positive results and make NASBA suitable for more POC diagnostic applications. Then we compare three different oligonucleotide sets to examine (1) the inhibitory effect of dimer formations, (2) false positives with poorly designed primers, and (3) the effect of beacon target location during real-time NASBA

SUBMITTER: Morabito K 

PROVIDER: S-EPMC7099912 | biostudies-literature | 2013 Jun

REPOSITORIES: biostudies-literature