Extremely sensitive, background-free gene detection using binary probes and beta replicase.
We have developed a specific and sensitive nucleic acid amplification assay that is suitable for routine gene detection. The assay is based on a novel molecular genetic strategy in which two different RNA probes are hybridized to adjacent positions on a target nucleic acid and then ligated to form an amplifiable reporter RNA. The reporter RNA is then replicated up to a hundred billion-fold in a 30-min isothermal reaction that signals the presence of the target. The assay can detect fewer than 100 nucleic acid molecules; it provides quantitative results over a wide range of target concentrations and it employs a universal format that can detect any infectious agent.
Tyagi
S
S
Department of Molecular Genetics, Public Health Research Institute, New York, NY 10016, USA.
Landegren
U
U
Tazi
M
M
Lizardi
P M
PM
Kramer
F R
FR
D001483:Base Sequence; D002460:Cell Line; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D005814:Genes, Viral; D005821:Genetic Techniques; D015497:HIV-1 / Q000201:enzymology / Q000235:genetics* / Q000502:physiology; D006801:Humans; D008214:Lymphocytes; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011777:Q beta Replicase; D015347:RNA Probes; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D012367:RNA, Viral / Q000032:analysis / Q000096:biosynthesis; D012680:Sensitivity and Specificity; D013698:Templates, Genetic; D014158:Transcription, Genetic
D015347:RNA Probes; D012333:RNA, Messenger; D012367:RNA, Viral; D011777:Q beta Replicase; D012321:DNA-Directed RNA Polymerases