ProAmpHT is high-throughput loop-mediated isothermal amplification (LAMP) run on the Optigene Genie® HT — a 96-well benchtop instrument that turns out fluorescent results in 15–30 minutes at a single temperature (typically 65°C). It is powered by the engineered GspSSD2.0 strand-displacing polymerase (DNA and RNA, no separate RT step) and confirmed by an automated anneal-curve read that rules out primer-dimer artefacts. It scales the same chemistry that runs on the field-portable Genie® II and III — same kits, same assay design — to centralized-lab throughput.
Key Facts
- 96 reactions per run on the Genie HT vs. 8 (Genie II) or 16 (Genie III) on the portable platforms.
- 15–30 minute time-to-result at a single isothermal temperature (60–68°C window; 65°C typical).
- GspSSD2.0 polymerase — strand-displacing, RT-capable, high inhibitor tolerance, lyophilization-stable.
- Anneal-curve confirmation after amplification provides sequence-specific verification analogous to a melt curve in qPCR.
- Applications include arbovirus surveillance (WNV, Zika, dengue), SARS-CoV-2, AMR gene profiling, food/water pathogens, and plant pathology.
- Pro-AmpRT WNV (Pro-Lab Diagnostics) is one example IVD/RUO LAMP kit validated on the Genie® family.
Why isothermal amplification matters
Conventional PCR has been the workhorse of molecular diagnostics for three decades, but it depends on a thermal cycler stepping through denaturation, annealing, and extension temperatures — typically 40+ cycles consuming 60–120 minutes of run time, plus the optical block and ramp hardware that make benchtop qPCR instruments expensive to deploy outside a centralized lab.
Loop-mediated isothermal amplification (LAMP), first described by Notomi et al. in 2000, sidesteps the thermal-cycling requirement entirely. Four-to-six primers recognize six-to-eight distinct regions of the target, and a strand-displacing polymerase produces a cauliflower-like concatemer at a single constant temperature. The reaction is fast (15–30 minutes), inhibitor-tolerant (it runs from crude lysates that would kill a Taq-based PCR), and can be read by simple fluorescence or even by eye via turbidity or colorimetric pH indicators.
ProAmpHT is the high-throughput productization of this chemistry: the Optigene-developed reagents, the Genie® HT instrument hardware, and the Genie Explorer software stack, integrated as a single workflow for laboratories that need to push hundreds of samples a day through a sequence-confirmed isothermal assay.
The GspSSD2.0 polymerase advantage
The polymerase is what makes or breaks an isothermal assay. ProAmpHT uses GspSSD2.0, an engineered variant of the strand-displacing DNA polymerase originally isolated from Geobacillus sp. The 2.0 generation was optimized along three axes that matter directly for clinical and field use:
- Inhibitor tolerance. GspSSD2.0 retains activity in the presence of heme, humic acids, polysaccharides, and the SDS/guanidine carryover that defeats Taq-based qPCR. This is what enables crude-lysate workflows — a brief heat lysis of saliva, swab eluate, mosquito homogenate, or a food rinsate can be added directly to the master mix.
- Reverse-transcription activity. The same enzyme molecule reverse-transcribes RNA and extends DNA, so RNA targets (SARS-CoV-2, West Nile virus, influenza) do not require a separate RT enzyme or an upstream cDNA step. One reaction tube, one buffer, one enzyme.
- Thermal stability and lyophilization. The enzyme is stable across the 60–68°C LAMP window and survives lyophilization in the master mix, which is what allows ambient-temperature shipping of dried-down kits for field use.
Coupled with Optigene's LAMP primer design conventions and the dual-labelled isothermal master mix, GspSSD2.0 makes it practical to convert an existing qPCR assay to an isothermal format and run it on the same crude sample input without re-engineering the upstream extraction.
Genie® HT vs. Genie® II / Genie® III throughput
The three Genie® platforms share chemistry and software — an assay developed on a portable Genie III in a public-health truck runs unchanged on a Genie HT in a centralized regional lab. The difference is form factor and throughput:
| Instrument | Wells per run | Form factor | Typical use |
|---|---|---|---|
| Genie® II | 8 (1 × strip) | Battery / mains, 2.3 kg | Point-of-need, field, single-sample triage |
| Genie® III | 16 (2 × strip) | Battery / mains, 2.4 kg | Field surveillance, on-farm, mosquito-trap reads |
| Genie® HT | 96 (1 × plate) | Benchtop, mains | Centralized lab, batched clinical or food testing |
Per-well chemistry is identical, so validation data and limit-of-detection figures transfer directly between platforms. A regional reference lab can develop and validate an assay on a single Genie III, then scale the same kit onto Genie HT plates without re-validating primer mixes or master-mix concentrations.
science Request Quote Optigene Genie® HT High-Throughput LAMP Instrument 96-well plate format, ProAmpHT chemistry, anneal-curve confirmation, Genie Explorer software. Sold and supported by Pro-Lab Diagnostics in Georgetown, TX. arrow_forwardAnneal-curve confirmation — the missing piece in older LAMP
Historically, the main scientific objection to LAMP has been specificity. With 4–6 primers in a single tube at one temperature, primer-dimer and non-specific amplification can produce a fluorescent signal that looks indistinguishable from real target amplification on the time-to-positive (Tp) trace alone.
ProAmpHT addresses this with an automated anneal-curve step at the end of each run. After the isothermal phase completes, the block ramps slowly downward while fluorescence is monitored continuously. Real concatemer amplicon re-anneals at a characteristic temperature (Ta) determined by its GC content and length, and that anneal event appears as a peak in the negative first derivative of the fluorescence trace — functionally analogous to a qPCR melt curve.
Each Optigene assay is published with an expected Ta window. The Genie Explorer software automatically calls a result positive only if both (a) the time-to-positive falls within the assay's expected window and (b) the anneal temperature matches the expected Ta ± tolerance. This two-parameter call dramatically reduces false positives from primer-dimer or carry-over contamination, and it satisfies the sequence-specificity expectations that clinical reviewers and regulators apply to nucleic-acid amplification tests.
Applications in practice
Arbovirus surveillance
Mosquito-borne flaviviruses are the canonical ProAmpHT use case. The Pro-Lab Diagnostics Pro-AmpRT WNV kit pairs RT-LAMP chemistry with an anneal-confirmed readout for West Nile virus detection directly from pooled mosquito homogenates collected with a standard mosquito collection & homogenization kit. A vector-control district can process trap-night pools on a Genie III at the field office and reflex confirmation onto a Genie HT in the regional lab. The same chemistry transfers to Zika and dengue with primer-set substitution.
SARS-CoV-2 and respiratory viruses
RT-LAMP for SARS-CoV-2 was deployed widely from mid-2020 onward, and the ProAmpHT format has continued to be used for surge testing, wastewater surveillance, and point-of-care screening where qPCR turnaround would be prohibitive. Crude-lysate workflows pair well with extraction-free saliva input — or with the Pro-Mag magnetic-bead or Pure Pro-Spin column extraction kits and the automated Mag Pro-32 instrument when classical RNA cleanup is preferred. The same workflow supports influenza A/B and RSV LAMP assays.
Antimicrobial-resistance (AMR) gene profiling
Isothermal amplification has become an attractive front-line tool for AMR gene detection because resistance markers (mecA, vanA/vanB, carbapenemase genes such as blaKPC, blaNDM, blaOXA-48) are short, well-conserved targets ideally suited to LAMP primer design. The 96-well throughput of the Genie HT makes it practical to run a multi-target AMR panel on every isolate flagged by routine culture, with results in under an hour rather than the next morning. DNase/RNase-free PCR plates are stocked alongside the instrument for routine plate-format runs.
Food, water, and plant pathology
Outside the clinical lab, ProAmpHT is used for Salmonella, Listeria, E. coli O157, and norovirus testing in food rinsates; for plant-pathology surveys of seed lots and orchard samples; and for environmental monitoring of recreational and drinking water. Inhibitor tolerance is the deciding factor — samples that would clog a qPCR instrument run cleanly on GspSSD2.0.
"Isothermal amplification is no longer the ‘poor relation’ of PCR. With anneal-curve confirmation and the GspSSD2.0 polymerase, ProAmpHT meets the specificity bar that clinical and food-safety reviewers expect, while keeping the sample-to-answer time under 30 minutes."
Getting started with ProAmpHT
For laboratories evaluating the platform, the typical path is: (1) define one or two priority targets, (2) order an Optigene assay kit or a Pro-Lab IVD/RUO kit such as Pro-AmpRT WNV, (3) run a small validation panel on a portable Genie II or III, then (4) scale to the Genie HT once the assay performance is locked. Pro-Lab Diagnostics supports the full workflow from extraction reagents (Pro-Mag, Pro-Spin) through automated cleanup (Mag Pro-32) to the Genie® instruments and assay kits themselves.
Frequently Asked Questions
What is ProAmpHT isothermal amplification?
ProAmpHT is the high-throughput implementation of loop-mediated isothermal amplification (LAMP) on the Optigene Genie® HT instrument. It uses GspSSD2.0 strand-displacing polymerase to amplify nucleic acids at a single constant temperature (typically 65°C), producing a fluorescent readout in 15–30 minutes across up to 96 wells in parallel, with automated anneal-curve confirmation.
How does the Genie HT differ from Genie II and Genie III?
Genie II (8 wells) and Genie III (16 wells) are portable battery-capable field instruments for point-of-need testing. The Genie HT is a benchtop 96-well plate instrument for centralized labs that need batched throughput. All three share the same chemistry, anneal-curve confirmation step, and Genie Explorer software — so assays transfer between platforms without revalidation.
What is the GspSSD2.0 polymerase advantage?
GspSSD2.0 is an engineered strand-displacing DNA polymerase that combines high inhibitor tolerance (crude lysates), reverse-transcription activity for RNA targets (no separate RT step), and thermal/lyophilization stability suitable for ambient-shipped dried-down kits. One enzyme handles both DNA and RNA amplification in a single tube.
How does anneal-curve confirmation reduce false positives?
After amplification, the block ramps down slowly while fluorescence is monitored. The amplicon re-anneals at a characteristic temperature (Ta) set by its GC content and length, producing a peak in the negative first derivative — analogous to a qPCR melt curve. Genie Explorer calls a positive only if both the time-to-positive and the observed Ta match the assay's expected windows, dramatically reducing false positives from primer-dimer.
What applications use ProAmpHT today?
Arbovirus surveillance (West Nile, Zika, dengue) directly from mosquito pools; SARS-CoV-2 and respiratory virus screening from saliva or swabs; antimicrobial-resistance gene profiling on cultured isolates (mecA, vanA/B, blaKPC, blaNDM, blaOXA-48); food and water pathogen testing; and plant pathology.
Does ProAmpHT replace qPCR?
It complements rather than replaces. qPCR remains the reference for quantitative viral-load and gene-expression work where Ct-based quantification is essential. ProAmpHT wins where speed, inhibitor tolerance, and instrument deployability matter more than quantitation — surveillance, screening, point-of-need, and triage workflows.
For technical questions about ProAmpHT, the Genie® HT, or any of the Pro-Lab molecular reagents, contact info@pro-lab.us or visit the Optigene Genie® product page for quote and configuration details.