Figure 3. Hybridization Protection Assay - Hologic Panther System Operator's Manual

1. The system moves the MTU into the HPA load station. The pipettor dispenses Probe
Reagent into each tube and the station mixes the tube contents.
The Probe Reagent contains AE-labeled oligonucleotides. The sequence of the
probes specifically complements the target amplicon.
If present, the amplicon hybridizes to the probes.
2. The MTU moves to the High Temperature Incubator for hybridization.
3. Once hybridized, the MTU moves to the HPA load station for addition of Selection
Reagent and mixing.
4. The MTU moves back to the High Temperature Incubator for the selection process.
During the selection phase, the hybridized and unhybridized probes are
differentiated by inactivating the AE label on the unhybridized probes. During
detection, the chemiluminescent signal produced by the hybridized probe is
measured in a luminometer and is reported as relative light units (RLUs). Inactive AE
does not emit light in the detection process.
5. The system dispenses Auto Detect 1 and Auto Detect 2 into the MTU. Auto Detect 1
and 2 react with the AE molecule, emitting a photon, or light signal.
6. A luminometer reads the light signal, if present.
7. After the luminometer reads the AE molecule signal, the MTU moves to the Output
Queue. The system injects sodium hypochlorite and Buffer for Deactivation into the
MTU to destroy any amplicon present. The MTU remains in the queue for 15 minutes
to decontaminate. The system then aspirates the fluids to a Liquid Waste Container
and deposits MTUs, pipette tips, and tiplets in a Solid Waste Container.
®
Panther System Operator's Manual
AW-17791-001 Rev. 002 (EN)

Figure 3. Hybridization Protection Assay

Overview
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