What is reverse hybridization?

Answer 1

Reverse hybridization assays offer a platform for highly specific probe hybridization.

Specific DNA probes are immobilized on a solid carried, such as nitrocellulose strips or Luminex beads.

The test procedure comprises three parts:

• Isolation of the DNA from the sample (reagents NOT provided in the kit)
• Amplification of target DNA using PCR
• Detection of the biotinylated product in our reverse hybridization assays

DNA or RNA derived amplification products can be denatured and hybridized to the immobilized probes. After stringent washing steps, the specific hybrids can be detected. On reverse hybridization strips, this results in a visible hybridization pattern. In the Luminex assay format, this can be measured by a specific Luminex reader.

Reverse hybridization assays have the following advantages:

• Hybridization is highly specific, allowing single nucleotide mismatch detection
• Detection is very sensitive, especially to detect minority species amplimers e.g. in mixed infections
• The read-out can be performed manually or automated
• The test is fast (about 8 hours, including amplification)