|Cervista HPV DNA|
Cervista HPV (Third Wave Technologies, Hologic, Inc, Madison, WI)Two assays are available:1. Cervista HPV HR Screening Test14 HR-HPV Types: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, or 682. Cervista HPV 16/18Identifies and differentiates HR Types 16 and 18, referred to as HPV-16/18 genotyping testIn the Cervista HPV DNA assays, two hybridizations and an enzyme cleavage occur in a primary reaction. The secondary reaction is detection with signal amplification using fluorescence resonance energy transfer (FRET). Prior to hybridization, the HPV DNA (if present) is isolated, denatured, and added to microplate wells.
|Cervista HPV DNA Secondary Reaction|
In the secondary reaction, the cleaved 5' Flaps attach to a FRET oligonucleotide probe. The FRET probe is curved such that the 5' Flap end nucleotide base pair attaches to the FRET probe in two places. Another cleavage reaction cleaves the 5' Flaps off the FRET probes and excites the fluorophores. The 5' Flaps reanneal and a cycling of the 5' Flaps off and on the FRET probes occurs. The emitted light is detected in a fluorescence plate reader. Because of the increasing number of 5' Flaps created in the primary reaction and the cycling of the 5' Flaps off and on the FRET probes, the target-specific signal is consequently amplified. During a 4-hour testing period, on average, the two reactions result in a 1-10 million-fold signal amplification.