HC2 High-Risk HPV DNA Test Cost & Procedure

The digene HC2 HPV DNA Test is a qualitative chemiluminescence-based in vitro microplate assay for the detection of 18 different types of human papillomavirus (HPV) DNA in cervical specimens. It is based on signal-amplified nucleic acid hybridization. The digene HC2 HPV DNA Test can distinguish between two forms of HPV DNA: low-risk and high-risk.

The 13 HR-HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, or 68 are detected by the Digene High-Risk HPV HC2 DNA test (Qiagen, Germantown, MD).

Method

The High-Risk HPV HC2 DNA test makes use of nucleic acid hybridization, signal amplification, and signal detection by chemiluminescence. A microplate is used to conduct and monitor hybridization and amplification procedures.

Step 1: Denaturing isolated DNA - If HPV DNA is present, it is denatured by the addition of a base solution to a liquid sample of cervical cells. The dsDNA is transformed into ssDNA strands by denaturation.

Step 2: of the hybridization process adds strands of HPV RNA sequences (RNA probes) that are complementary to HR-HPV types. By linking complimentary bases with the ssDNA, the probes anneal or hybridise. DNA:RNA hybrids can be produced.

Each of the following diagrams (Hybrid Capture 2 and Cervista) artificially constructs the DNA target sequence to represent base pairing, hybridization, and detection. There would also be longer actual target sequences and probes.

Step 3: RNA:DNA hybrids were dissolved and put to a microplate well coated with capture antibodies designed specifically for RNA:DNA hybrids. The antibodies bind to and adhere to RNA:DNA hybrids, which are then trapped in the microplate wells.

Step 4: Addition of various alkaline phosphatase-conjugated antibodies for RNA:DNA hybrids. The tagged antibodies create sandwiches when they bind to the hybrids. Each sandwich has three parts: a capture antibody, antibody with a label: hybrid.

To get rid of any unbound antibodies that were alkaline phosphatase-labeled, the microplate is washed. Each hybrid is bound by numerous tagged antibodies, which causes a 3000 fold increase in signal.

In the microplate wells of the negative samples and the negative control, unbound alkaline phosphatase-labeled antibodies will remain if the wash step is skipped. They will create chemiluminescence when the substrate is introduced, along with the positive specimen and the positive control..

Step 5: A substrate for chemiluminescence is applied. Enzymes and substrates undergo a chemical reaction that produces light, and a luminometer detects the light.