Home » Technology Platforms » Reverse Hybridization Assay (RHA)

Reverse Hybridization Assay (RHA)

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 LMNX 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 LMNX assay format, this can be measured by a specific LMNX 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)

Strip-based reverse hybridization

Probes can be immobilized on nitrocellulose strips as parallel lines. Each line represents a specific probe. After hybridization and stringent wash, specific hybrids can be detected by colorimetric reactions. By visually comparing the hybridization pattern on the strip to a reference read-out template, the test result can easily be deduced.

LMNX testing format

The LMNX platform is based on polystyrene color-coded microspheres. Each bead type can be coated with a specific reagent. This permits the specific capture and detection of specific analytes from a biological sample. Within the LMNX reading instrument, lasers excite the internal dyes that identify each microsphere particle, carrying a specific reagent into a specific bead type.
A second reporter dye can be generated upon reactivity of the biological analyte captured from the biological sample during the assay. Multiple readings are made on each bead set. In this way, LMNX microsphere technology allows multiplexing assays of up to 100 unique analytes within a single sample.

Contact us

Just give us a call, send us an e-mail or make an appointment!