title: 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 tuberculosis.
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.
authors.affiliation: Clinical Experimental Laboratory of Biosensor and Microarray, and Center of Molecular and Genetic Diagnosis, Southwest Hospital, 3rd Military Medical University, Chongqing, PR China.
authors.forename: Qinghai
authors.initials: Q
authors.lastname: Chen
authors.forename: Nan
authors.initials: N
authors.lastname: Wu
authors.forename: Meng
authors.initials: M
authors.lastname: Xie
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
authors.forename: Ming
authors.initials: M
authors.lastname: Chen
authors.forename: Jianjun
authors.initials: J
authors.lastname: Li
authors.forename: Lisha
authors.initials: L
authors.lastname: Zhuo
authors.forename: Hong
authors.initials: H
authors.lastname: Kuang
authors.forename: Weiling
authors.initials: W
authors.lastname: Fu
mesh_terms: D001483:Base Sequence; D019921:Biotinylation; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D057166:High-Throughput Screening Assays / Q000379:methods*; D015335:Molecular Probes / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000235:genetics*; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry* / Q000235:genetics; D017354:Point Mutation / Q000235:genetics*
chemical_list: D015335:Molecular Probes; D015345:Oligonucleotide Probes