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

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

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 SLI...

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BACKGROUND Here we present a novel promising microbial diagnostic method that combines the sensitivity of Nucleic Acid Sequence Based Amplification (NASBA) with the high information content of microarray technology for the detection of bacterial tmRNA molecules. The NASBA protocol was modified to include aminoallyl-UTP (aaUTP) molecules that were incorporated into nascent RNA during the NASBA r...

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BACKGROUND The hybridization of synthetic Streptococcus pneumoniae tmRNA on a detection microarray is slow at 34 degrees C resulting in low signal intensities. RESULTS We demonstrate that adding specific DNA helper oligonucleotides (chaperones) to the hybridization buffer increases the signal strength at a given temperature and thus makes the specific detection of Streptococcus pneumoniae tmR...

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Molecular beacon probes combined with amplification by NASBA enable homogeneous, real-time detection of RNA.

Molecular beacon probes can be employed in a NASBA amplicon detection system to generate a specific fluorescent signal concomitantly with amplification. A molecular beacon, designed to hybridize within the target sequence, was introduced into NASBA reactions that amplify the genomic RNA of potato leafroll virus (PLRV). During amplification, the probe anneals to the antisense RNA amplicon genera...

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ژورنال

عنوان ژورنال: BMC Biotechnology

سال: 2011

ISSN: 1472-6750

DOI: 10.1186/1472-6750-11-17