Category Archive

Below you'll find a list of all items that have been categorized as “HIV Drug Resistance”

PANDAA HIVDR Evaluations

Join the international discussion on how focussed genotyping by PANDAA can facilitate universal access to HIV drug resistance testing. 

Focused Genotyping for HIV Drug Resistance

PANDAA HIVDR is a collection of distinctive focused genotyping assays that are uniquely designed to provide accurate and rapid diagnostic information relevant to the management of HIV drug resistance.

PANDAA HIVDR is a collection of distinctive focused genotyping assays that are uniquely designed to provide accurate and rapid diagnostic information relevant to the management of HIV drug resistance.

PANDAA HIVDR are real-time PCR assays that are available in multiple formats:

  • Two pre-defined panels—ReFocus I and ReFocus II—for patients failing an NNRTI- or PI-based regimen, respectively
  • ReFocus Select, a collection of individual multiplex assays that allows you to develop a customised focused genotyping panel based on the drug resistance mutations of interest.
PANDAA HIVDRHIV TargetsDrug Resistance Mutations
ReFocus IReverse Transcriptase65R, 103NS, 106M, 181C, 184VI, 190A
ReFocus IIProtease47AV, 50L, 76V, 82AFT, 84V, 88S
Reverse Transcriptase65R, 184VI
ReFocus SelectReverse Transcriptase65R, 184VI, 103N, 103NS, 106M, 181C, 190A
Protease47AV, 47A, 47V, 50L, 76V, 82AFT, 82A, 82F, 82T, 84V, 88S
Integrase118R, 148H, 148K, 148R, 155H, 263K

If more than one resistance-conferring mutation is possible at a codon e.g., M184VI in reverse transcriptase, the ReFocus I and II assays do not differentiate between the DRMs. Using the customisable panels with ReFocus Select, you do have the option to differentiate DRMs, for example, 82A, 82F and 82T DRMs in protease can be genotyped separately or together with the 82AFT PANDAA.

Inquiry

Complete the form below or email change@aldatubio.com

Download our PANDAA HIVDR brochure:
Focused HIVDR Genotyping Assays by Aldatu.pdf

Discover PANDAA HIVDR

Multiplexing enables high-throughput resistance genotyping.

Each PANDAA HIVDR reaction contains amplification and detection reagents for two DRMs. For example, 65R and 181C are genotyped together in a single reaction using the ReFocus I panel. Each PANDAA HIVDR reaction also contains a viral quantifier (VQ) that targets a conserved region of the HIV-1 genome and is used as a proxy for total HIV-1 nucleic acid to determine the relative abundance of each DRM. In four hours, up to 24 patients can be genotyped using ReFocus I and 20 patients with ReFocus II, after sample extraction. A custom panel of twelve DRMs [six PANDAA HIVDR reaction] using ReFocus Select will genotype up to 12 patients in same time.

Focused resistance genotyping for 20 patients can be completed within four hours.

PANDAA PreAmp is a three-step process: after adding the Stop-Start enzyme to the PreAmp buffer, the complete mastermix is added to each processed sample. Once pre-amplification is complete, the sample can be stored at 4°C or used immediately for PANDAA HIVDR genotyping.

 

PANDAA PreAmp

PANDAA HIVDR genotyping setup takes ~30 minutes with a real-time PCR run time of ~70 minutes.

Data analysis is performed in <10 minutes. The amplification mastermix [PRxB] is added directly to the tubes containing each of the PANDAAs [our resistance genotyping reagents]. Each PANDAA + PRxB mix is used to fill a real-time PCR plate. PreAmp sample [or control] is added to one of each of the HIVDR genotyping wells and no mixing is necessary. Any excess PANDAA + PRxB mix can be stored at 4°C for several weeks for use with additional samples.

 

PANDAA HIVDR Setup

PANDAA PreAmp allows low viral load samples to undergo focused genotyping for several DRMs without any loss in sensitivity.

PreAmp is a rigorously validated RT-PCR-based approach for the consistent and unbiased amplification of the HIV pol gene. By restricting pre-amplification to ten cycles with a limited concentration of amplification reagents,

 

PreAmp generates sufficient product from low copy RNA samples for multiple downstream PANDAA HIVDR assays, while minimizing SNP frequency biasing and cross-contamination that could result from over-amplification. PreAmp generates sufficient sample material for >40 PANDAA HIVDR assays or can be used for other downstream application such as Sanger sequencing. The PreAmp module contains the PreAmp amplification buffer, Stop-Start enzyme (a specially formulated hot-start reverse transcriptase) and PreAmp Primers, a reagent mix that amplifies regions 2252 → 5073 of the HIV-1 genome.

 

PANDAA PreAmp can be stored for several weeks at 4°C with no loss in sensitivity, and no sample cleanup is required before using PANDAA PreAmp in downstream applications.

Multiplexing enables high-throughput resistance genotyping.

Each PANDAA HIVDR reaction contains amplification and detection reagents for two DRMs. For example, 65R and 181C are genotyped together in a single reaction using the ReFocus I panel. Each PANDAA HIVDR reaction also contains a viral quantifier (VQ) that targets a conserved region of the HIV-1 genome and is used as a proxy for total HIV-1 nucleic acid to determine the relative abundance of each DRM. In four hours, up to 24 patients can be genotyped using ReFocus I and 20 patients with ReFocus II, after sample extraction. A custom panel of twelve DRMs [six PANDAA HIVDR reaction] using ReFocus Select will genotype up to 12 patients in same time.

Focused resistance genotyping for 20 patients can be completed within four hours.

PANDAA PreAmp is a three-step process: after adding the Stop-Start enzyme to the PreAmp buffer, the complete mastermix is added to each processed sample. Once pre-amplification is complete, the sample can be stored at 4°C or used immediately for PANDAA HIVDR genotyping.

 

PANDAA PreAmp

PANDAA HIVDR genotyping setup takes ~30 minutes with a real-time PCR run time of ~70 minutes.

Data analysis is performed in <10 minutes. The amplification mastermix [PRxB] is added directly to the tubes containing each of the PANDAAs [our resistance genotyping reagents]. Each PANDAA + PRxB mix is used to fill a real-time PCR plate. PreAmp sample [or control] is added to one of each of the HIVDR genotyping wells and no mixing is necessary. Any excess PANDAA + PRxB mix can be stored at 4°C for several weeks for use with additional samples.

 

PANDAA HIVDR Setup

PANDAA PreAmp allows low viral load samples to undergo focused genotyping for several DRMs without any loss in sensitivity.

PreAmp is a rigorously validated RT-PCR-based approach for the consistent and unbiased amplification of the HIV pol gene. By restricting pre-amplification to ten cycles with a limited concentration of amplification reagents,

 

PreAmp generates sufficient product from low copy RNA samples for multiple downstream PANDAA HIVDR assays, while minimizing SNP frequency biasing and cross-contamination that could result from over-amplification. PreAmp generates sufficient sample material for >40 PANDAA HIVDR assays or can be used for other downstream application such as Sanger sequencing. The PreAmp module contains the PreAmp amplification buffer, Stop-Start enzyme (a specially formulated hot-start reverse transcriptase) and PreAmp Primers, a reagent mix that amplifies regions 2252 → 5073 of the HIV-1 genome.

 

PANDAA PreAmp can be stored for several weeks at 4°C with no loss in sensitivity, and no sample cleanup is required before using PANDAA PreAmp in downstream applications.

Inquiry