Journal Article

Detection of NASBA amplified bacterial tmRNA molecules on SLICSel designed microarray probes

Parkel, Sven, Palta, Priit, Toome, Kadri, Maher, Majella, Kurg, Ants, Scheler, Ott, Barry, Thomas, Kaplinski, Lauris, Glynn, Barry, Remm, Maido

2011

Overview

Background We present a comprehensive technological solution for bacterial diagnostics using tmRNA as a marker molecule. A robust probe design algorithm for microbial detection microarray is implemented. The probes were evaluated for specificity and, combined with NASBA (Nucleic Acid Sequence Based Amplification) amplification, for sensitivity. Results We developed a new web-based program SLICSel for the design of hybridization probes, based on nearest-neighbor thermodynamic modeling. A SLICSel minimum binding energy difference criterion of 4 kcal/mol was sufficient to design of Streptococcus pneumoniae tmRNA specific microarray probes. With lower binding energy difference criteria, additional hybridization specificity tests on the microarray were needed to eliminate non-specific probes. Using SLICSel designed microarray probes and NASBA we were able to detect S. pneumoniae tmRNA from a series of total RNA dilutions equivalent to the RNA content of 0.1-10 CFU. Conclusions The described technological solution and both its separate components SLICSel and NASBA-microarray technology independently are applicative for many different areas of microbial diagnostics.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Analysis / Applied Microbiology / Bacteria / Bacterial infections / Biochemical Engineering / Biomedical Engineering/Biotechnology / Biotechnology / Chemistry / Chemistry and Materials Science / Design / Diagnosis / DNA microarrays / Genes / Genetic algorithms / Genetic Engineering / Hybridization / Methodology / Methodology Article / Methods / Microbiology / Nucleic acid probes / Nucleic acids / Oligonucleotide Array Sequence Analysis - methods / Plant Breeding/Biotechnology / Probes / RNA Probes - genetics / RNA, Bacterial - genetics / Self-Sustained Sequence Replication - methods / Software / Species Specificity / Streptococcus infections / Streptococcus pneumoniae - genetics