The Reliance SARS-CoV-2 RT-PCR Test provides real-time PCR detection of multiple RNA targets from the SARS-CoV-2 virus with great sensitivity and speed. It is designed to detect nucleic acid from SARS-CoV-2 in nasopharyngeal swabs specimen from individuals suspected of COVID-19 by their healthcare provider.
More FAQs: Reliance SARS-CoV-2 RT-PCR Assay Kit
Kit Contents, Storage, and Use
Can the Reliance SARS-CoV-2 RT-PCR Assay Kit be run on a Research Use Only (RUO) instrument?
A U.S. FDA Emergency Use Authorization (EUA) is a special use category due to an FDA-declared public health emergency pandemic, where an RUO kit is temporarily allowed for use as a diagnostic during the time period that the EUA is permitted by the FDA. Once the FDA-declared public health emergency is lifted, EUA use is no longer permitted.
Can the kit be used in a non-clinical lab for non-diagnostic purposes?
Using this EUA-approved kit for non-diagnostic purposes, such as surveillance, is an off-label use and is not covered under the guidelines of the EUA.
Can kit components undergo multiple freeze-thaw cycles or be stored at 4–8°C to avoid freeze-thaw?
All kit components should be stored at –20°C. Reliance One-Step Multiplex RT-qPCR Supermix and the SARS-CoV-2 RT-PCR Oligos can undergo up to 5 freeze/thaw cycles. The Exact Diagnostics SARS-CoV-2 Standard and Negative are labeled as “single use”. If desired, the Standard and Negative controls can be thawed, aliquoted into smaller volumes, and stored frozen.
Is the Reliance SARS-CoV-2 RT-PCR Assay Kit compatible with saliva samples?
The kit is designed to detect RNA from the SARS-CoV-2 virus in nasopharyngeal swabs, oropharyngeal swabs, nasal swabs, anterior nasal swabs, mid-turbinate nasal swabs, nasal aspirates, and nasal washes from patients who are suspected of COVID-19. The use of this kit with saliva samples has not been validated.
Can customers choose to use other extraction methods or kits not mentioned in the IFU?
Performance of this kit has been validated with samples extracted using the Thermo Fisher MagMax Viral/Pathogen Nucleic Acid Isolation Kit and the QIAGEN QIAamp Viral RNA Mini Kit. Currently, the EUA requires use of one of these kits.
Can labs use an automated RNA extraction method to extract samples before running the kit?
This kit is only validated for two manual sample extraction methods: Thermo Fisher MagMAX Viral/Pathogen Nucleic Acid Isolation Kit and QIAGEN QIAamp Viral RNA Mini Kit. Use of this kit with other extraction methods is not covered under the current EUA. Automated extraction (on QIAcube or Kingfisher) using these kits is supported by the external manufacturer’s procedure and requires customer’s own validation.
Where can more information about the Exact Diagnostics controls used in the kit be found?
The molecular controls from Exact Diagnostics were developed independently of the Reliance SARS-CoV-2 RT-PCR Assay Kit. The Standard and Run Controls have been quantitated using digital PCR. The Standard is used as a reference material for research testing of the novel coronavirus allowing laboratories to validate assays. Additionally, the Standard can be validated for use by the laboratory as an external run control. Routine use of the Run Controls allow laboratories to evaluate the day-to-day and lot-to-lot variation of their molecular assay and test for operator proficiency.
For more information on Exact Diagnostic controls, please visit the product pages on Bio-Rad.com. Customers can also order the controls separately or obtain the CoA for the controls here. Note that the size of the controls sold separately is 5 x 0.3 ml.
Data Analysis and Interpretation
Do the mutations found in UK variant (B.1.1.7 lineage) affect the Reliance SARS-CoV-2 RT-PCR Assay?
The Reliance SARS-CoV-2 RT-PCR Assay is designed to identify the SARS-CoV-2 nucleoprotein (N) gene and not the spike protein (S) gene. Mutations found in the UK variant do not overlap the target region in this kit, so performance will not be affected by the variant.
As the virus continues to propagate and spread, additional mutations may be acquired in any region of the virus genome. Should a mutation be acquired in the N gene, the performance of the Reliance SARS-CoV-2 RT-PCR Assay Kit may be impacted.
Does Bio-Rad have a proactive Risk Management Plan?
Bio-Rad monitors SARS-CoV-2 variants in the NCBI and GISAID databases for reports of novel variants and routinely performs bioinformatic analysis to confirm in silico that the performance of the kit is not impacted by known mutations.
What are the guidelines for resolving inconclusive results?
An inconclusive result occurs when only one of the N gene targets is amplified and crosses the quantification threshold before cycle 40 and the plate controls product expected results. For samples with inconclusive results, re-test using residual extracted sample, if available. If results are still inconclusive or the amount of residual extracted sample is insufficient, re-extract the patient sample or collect another sample from the patient.
Why is the Limit of Detection (LoD) reported as a range (150 to 500 copies/ml)?
This range represents variation between instruments and is expected. The LoD range accounts for variance in detection quality and signal-to-background noise ratios between the instruments. In addition, clinical samples are comprised of a complex matrix and can be extracted using different methods, both of which contribute to overall variability.
Is data interpretation manual or automatic? Is positive or negative determination incorporated into the software?
Results are manually interpreted. On CFX Opus and Touch Systems, data analysis is performed using CFX Maestro v2.0 Software. The software displays Cq values based on manual threshold and baseline settings. Explicit instructions in how to interpret the data and determine if the sample is positive for SARS-CoV-2 are defined in the IFU.
Does the IFU provide instructions on how to interpret data generated on the AB7500 Fast Real-Time PCR System?
Yes, the IFU includes a section on data analysis using the AB7500 Software.
Real-time PCR or Droplet Digital PCR: which is the right technology to use?
Bio-Rad provides a full range of molecular testing options for COVID-19 testing, including both Real-Time PCR and Droplet Digital PCR (ddPCR). Real-time PCR is the gold standard in COVID-19 diagnostic testing, providing accessible, rapid results. The high sensitivity of ddPCR makes it well suited for population screening through pooled testing, and to aid in confirming negative results by other methods.