Solid-phase sequence scanning for drug resistance detection in tuberculosis.
DNA chip arrays hold considerable promise for diagnostic sequencing of polymerase chain reaction (PCR) products. To date, however, arrays have been relatively expensive, complex to use and difficult to interpret, preventing their adaptation to the clinical lab. A moderate density array method has been developed that enables efficient, easy-to-interpret and robust solid-phase PCR product sequencing. Here, the results of Mycobacterium tuberculosis rifampin resistance mutation detection by primer-extension-based sequence scanning of the rpo B gene of M. tuberculosis are presented. Rifampin resistant clinical isolates were identified in as little as 1 h post PCR amplification with visual results detection.
Head
S R
SR
Alpha Center, Orchid Biocomputer, Inc., Hopkins Bayview Research Campus, 5210 Eastern Avenue, Baltimore, Maryland, 21224, USA.
Parikh
K
K
Rogers
Y H
YH
Bishai
W
W
Goelet
P
P
Boyce-Jacino
M T
MT
D001426:Bacterial Proteins / Q000235:genetics*; D003062:Codon / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics*; D012321:DNA-Directed RNA Polymerases; D004352:Drug Resistance, Microbial / Q000235:genetics*; D005798:Genes, Bacterial; D005838:Genotype; D006801:Humans; D007091:Image Processing, Computer-Assisted; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010940:Plant Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012293:Rifampin / Q000494:pharmacology*; D017422:Sequence Analysis, DNA; D012984:Software; D013997:Time Factors; D014376:Tuberculosis / Q000188:drug therapy / Q000382:microbiology*
D001426:Bacterial Proteins; D003062:Codon; D017931:DNA Primers; D004269:DNA, Bacterial; D010940:Plant Proteins; C101305:RpoB protein, Oryza sativa; D012321:DNA-Directed RNA Polymerases; D012293:Rifampin