HIV 1 RNA

Background

HIV & ART

It is estimated that 1.2 million people over the age of 13 live with HIV in the US with 50,000 new HIV infections occurring every year (Prevalence of diagnosed and undiagnosed HIV infection-United States, 2015). Of the 86 % who have been diagnosed, 37% receive antiretroviral therapy (ART) (Vital Signs: HIV diagnosis, care and treatment among persons living with HIV-United States, 2011, 2014). Conventional ART involves a regimen of antiretroviral drugs (ARV) that optimally suppress the HIV viral load of the patient to stop advancement of the disease and is recommended as an initial therapy to HIV positive patients to conserve viral loads below the limit of detection (Treatment and Care).

ART Failure

The major contributor to ART failure is lack of therapy adherence, and occurs in approximately 5-10% of HIV patients per year. When ART failure happens, viral loads rise and drug resistance can develop. The development of HIV drug resistance means that certain ART regimens will no longer be effective for that patient, and that perhaps the patient could spread this resistance to their partners, i.e. transmitted drug resistance (TDR) (Tilghman, et al., 2014) (Attia, Egger, Muller, Zwhalen, & Low, 2009). More specifically, risk of HIV transmission and potential acquired/transmitted drug resistance increases when viral loads exceed 1000 viral copies/mL (Attia, Egger, Muller, Zwhalen, & Low, 2009). Thus, ART failure needs to be diagnosed quickly to prevent disease progression in that patient and to prevent transmission of the virus to a sexual or needle sharing partner (Loutfy, 2013) (WHO, Consolidated guidelines on the use of antiretroviral drugs for treating and preventing HIV infection, 2013)

ART Monitoring

Viral load tests are administered to ART receiving patients 2-3 times per year to monitor ART (Blair, Fagan, & Frazier, 2014). The current standard for viral load detection takes upwards of 1 week for a clinician to acquire an actionable result using a centralized lab. This delay is a time period where a patient with ART failure can transmit the virus and/or develop drug resistance. Laboratories, while using the most advanced technology to achieve appropriate sensitivity, have turna-round times (TATs) that are too long, from sample preparation and handling, to achieve clinical relevance that will best benefit the patient (Sanavio & Krol, 2015).

The Fluxergy Analyzer Beta and HIV-1 Viral RNA assay provide a rapid, cost-effective solution for HIV viral load detection without sample preparation. With immediate and reliable results in less than 30 minutes, a laboratory, hospital or clinic can speed up TATs such that a clinician can prescribe an appropriate regimen to a patient at the point of care and avoid the costly risks associated with infectious levels of HIV.

Key Benefits

Simple workflow

No sample preparation

Automated data interpretation

Sample to result in 30 minutes

Detection of viral RNA

Comprehensive coverage of subtypes and clades

Real-time monitoring of ART

Assay Specifications

Sample Type K3/ETDA (purple top) blood plasma
Species Human
Storage Condition Fluxergy Card - Room temperature ® ~23° C
Fluxergy Buffer - Frozen ® -20° C
Genotype Inclusivity Group M subtypes A-H, Group 0

Kit Contents

  • Fluxergy HIV-1 RNA Buffer
  • Fluxergy Card

Customer supplied reagents and supplies

  • Clinical sample for testing

Sample collection method

K3/EDTA plasma: Draw 1mL of venous whole blood in a purple top, K3/EDTA vacutainer tube. Centrifuge for at least 10 minutes at 1500-2500 RPM. Pipette the blood plasma into screw-cap vial and label. If not testing immediately store at 4°C, or at -20°C if more than 1 day. If thawing sample from frozen state, thaw on ice.

Testing Methods

Pipette 20uL of thoroughly mixed human blood plasma into Fluxergy HIV-1 Buffer 1. Vortex the sample and buffer for 5 seconds and pipette 100uL of the mixture into the Fluxergy Card. Insert the Fluxergy Card into the Fluxergy Analyzer Beta and begin test. (For detailed instructions, refer to Fluxergy Analyzer Beta instruction manual)

Intended use

The Fluxergy HIV-1 RNA assay is a qualitative test for the rapid detection of pathogenic HIV-1 in human blood plasma samples.

Positive PCR Result: indicates that RNA of the target organism is present in the tested sample.

Negative PCR Result: indicates that RNA of the target organism was not detected in the tested sample. However, a negative PCR results may also indicate that the number of target organisms is below the limit of detection.

Warnings and Precautions

  • Fluxergy's HIV-1 RNA assay is for Research Use Only (RUO). It is not intended for diagnostic use.
  • Fluxergy's HIV-1 RNA assay is compatible only with the Fluxergy Analyzer Beta device.
  • All specimens should be handled as potentially infectious agents and according to universal safety precautions.
  • This Fluxergy HIV-1 RNA assay is compatible only for K3/EDTA human blood plasma (sample type). The sample type and anticoagulant must be specified as each buffer is specially crafted for the sample. Fluxergy HIV-1 RNA assays for other anticoagulants (e.g. heparin, sodium citrate) are available upon request.
  • Contamination of the sample and kit contents may lead to erroneous results. Use aseptic technique and a clean workspace whenever possible.
  • Store Fluxergy assay kit at recommended storage temperature and conduct assay within specified environment (e.g. temperature and humidity) for optimal performance.
  • Follow appropriate specimen collection, storage and processing for optimal performance.
  • Use Fluxergy supplied swabs, reagents, pipettes and pipette tips to conduct assay for optimal performance.

Clinical Performance

Fluxergy HIV-1 RNA Assay versus NAAT

239 HIV-1 viral RNA positive and 26 HIV-1 viral RNA negative human blood plasma samples were collected for testing ofthe Fluxergy HIV-1 RNA Assay on the Fluxergy Analyzer Beta. Positive samples ranged from 103-107, viral RNA particles/mL. The samples were previously categorized by a 3rd party reference laboratory using a FDA approved, quantitative HIV-1 viral RNA assay.

Fluxergy Result Standard of Care Testing
Detected 239 0 239
Not Detected 0 26 26
Total 239 26 265
Positive Agreement (247/247) Negative Agreement (26/26)

Detection of Clinical Sample

Clinical sample detection. Clinical sample, specifically HIV-1 positive human blood plasma, was run on the Fluxergy Analyzer Beta using the Fluxergy HIV-1 RNA assay. Samples were previously categorized via a FDA approved, quantitative HIV-1 viral RNA assay by a 3rd party reference laboratory.

References

  1. Attia, S., Egger, M., Muller, M., Zwhalen, M., & Low, N. (2009). Sexual transmission of HIV according to viral load and antiretroviral therapy: a systematic review and meta-analysis. AIDS, 23,1397-404.
  2. Blair, J., Fagan, J., & Frazier, E. (2014). Behavioral and clinical characteristics of persons receiving medical care for HIV infection-Medical Monitoring Project, 2009. Morb Mortal Wkly Rep Surveill Summ, 63,1-22.
  3. (2015). Guidelines for the Use of Antiretroviral Agents in HIV-1 Infected Adults and Adolescents. Aids Info. Retrieved from guidelines
  4. Loutfy, M. (2013). Systematic review of HIV transmission between heterosexual serodiscordant couples where the HIV-positive partner is fully suppressed on antiretroviral therapy. PLoS One, 8.
  5. Prevalence of diagnosed and undiagnosed HIV infection-United States. (2015). MMWR Morb Mortal Wkly, 64, 657-62.
  6. Sanavio, B., & Krol, S. (2015). On the Slow Diffusion of Point-of-Care Systems in Therapeutic Drug Monitoring. Frontiers in Bioengineering and Biotechnology, 3, 20.
  7. Tilghman, M., Perez-Santiago, J., & Osorio, G. (2014). Community HIV-1 Drug Resistance is Associated with Transmitted Drug Resistance. HIV Medicine, 15, 339-46.
  8. Tilghman, M., Perez-Santiago, J., Osorio, G., Little, S., Richman, D., Mathews, W., ... Smith, D. (2014). Community HIV-1 Drug Resistance is Associated with Transmitted Drug Resistance. HIV Med, 15, 339-46.
  9. (n.d.). Treatment and Care. WHO. Retrieved from here.
  10. Vital Signs: HIV diagnosis, care and treatment among persons living with HIV-United States, 2011. (2014). MMWR Morb Mortal Wkly, 63,1113-7.
  11. WHO. (2013). Consolidated guidelines on the use of antiretroviral drugs for treating and preventing HIV infection.133-4.