摘要 |
Compositions and methods are provided for amplifying and or detecting nucleic acid targets using labeled polynucleotide probes and antiprobes that interact together and with complementary targets. Competitive thermodynamic interactions of these components result in signaling changes that indicate target frequency, and can provide error-checking functions that facilitate single base discrimination. These probe:antiprobe compositions enable real-time PCR detection, end-point detection and microarray detection of microbial species, drug resistant mutants, and cancer related variants. Some probe:antiprobe systems function as an internal probe, between two primers, and some function as a primer-probe. Some target sequences are discriminated or quantified by employing two probe:antiprobe systems to detect different aspects of the same template. Systems are also provided that enhance target amplification and detection for specific single base variants. A probe may also be modified by introducing a base mismatch to increase thermodynamic discrimination of a correct versus incorrect target differing by a single base. |