01 April 2012
A novel asymmetric-loop molecular beacon-based two-phase hybridization assay for accurate and high-throughput detection of multiple drug resistance-conferring point mutations in Mycobacterium tuberculosisQinghai ChenAG, Nan WuBC, Meng XieE, Bo ZhangC, Ming ChenF, Jianjun LiD, Lisha ZhuoF, Hong KuangD, Weiling FuA
Med Sci Monit 2012; 18(4): HY5-8
ABSTRACT: The accurate and high-throughput detection of drug resistance-related multiple point mutations remains a challenge. Although the combination of molecular beacons with bio-immobilization technology, such as microarray, is promising, its application is difficult due to the ineffective immobilization of molecular beacons on the chip surface. Here, we propose a novel asymmetric-loop molecular beacon in which the loop consists of 2 parts. One is complementary to a target, while the other is complementary to an oligonucleotide probe immobilized on the chip surface. With this novel probe, a two-phase hybridization assay can be used for simultaneously detecting multiple point mutations. This assay will have advantages, such as easy probe availability, multiplex detection, low background, and high-efficiency hybridization, and may provide a new avenue for the immobilization of molecular beacons and high-throughput detection of point mutations.
Keywords: Nucleic Acid Hybridization - methods, Nucleic Acid Conformation, Mycobacterium tuberculosis - genetics, Molecular Sequence Data, Molecular Probes - genetics, High-Throughput Screening Assays - methods, Drug Resistance, Multiple, Bacterial - genetics, Biotinylation, Base Sequence, Oligonucleotide Probes - genetics, Point Mutation - genetics
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