Testing Performed with Support from the NIH RADx® Tech Program
The World Health Organization (WHO) estimates that 50 million people globally have a chronic hepatitis C virus (HCV) infection. HCV is a bloodborne virus that affects the liver and can lead to lifelong illness including liver cirrhosis and cancer, which can be life-threatening. It is also estimated that in 2022, approximately 242,000 people died due to complications resulting from the virus. Though there is no vaccine for HCV, there are antiviral medicines that can cure more than 95 percent of people who are infected, but access to diagnosis and treatment is low. Early detection and resulting treatment can improve long-term health, potentially saving millions of lives.
The WHO has a goal of HCV elimination by 2030. To help achieve this goal, fast and effective diagnostics should be accessible, rapid, and non-invasive to reach populations with an increased risk for HCV infection. MRIGlobal collaborator Cepheid developed a molecular based diagnostic test for HCV ribonucleic acid (RNA) in human capillary blood through automated qualitative in vitro reverse transcription polymerase chain reaction (RT-PCR). The Cepheid Xpert® HCV is a CLIA-waived test with a run time of about one hour, and it has a small instrument footprint, making it ideal for point of care (POC) testing.
In support of this effort, MRIGlobal performed analytical and flex studies on the new HCV diagnostic test with support from the NIH Rapid Acceleration of Diagnostics (RADx®) Independent Test Assessment Program (ITAP). MRIGlobal has been a core part of ITAP since its inception, having supported companies seeking Emergency Use Authorization (EUA) of their COVID-19 at-home test devices, multiplex respiratory disease (COVID-19, Influenza A, Influenza B) at-home devices, and an mpox assay.
In these studies of the HCV diagnostic, we performed:
Limit of Detection (LoD)
- LoD range finding for 6 HCV genotypes was conducted using a minimum of 4 concentrations with 5 replicates per concentration.
- LoD confirmation studies were then performed with a minimum of 3 concentrations each with 20 replicates to determine the concentrations at which positivity rates of 60%-90%, 95%, and 100% were observed.
- From the LoD results, we calculated the Probit LoD using the Analyse-It software in Excel.
Flex Testing
Flex testing was performed across nine studies to determine points of test failure when the device is run outside of the provided Quick Reference Instructions (QRI) and Instructions for Use (IFU) and determine robustness of the test.
- Incorrect Handling (Mixing) of Sample: Samples were tested following various levels of mixing both immediately after loading into the microtainer and after letting the microtainer sit for 15 minutes.
- Incorrect Handling (Mixing) of External Control: Controls were tested following various levels of mixing.
- Incorrect Timing of Cartridge Preparation: Samples were added to the cartridge and tested after waiting 0 minutes, 15 minutes, and 30 minutes.
- Incorrect Test Volume: Different volumes of sample ranging from 25uL to 500uL were tested.
- Incorrect Handling of Cartridge: Samples were tested after mishandling the cartridges in various ways.
- Four Environmental Conditions (Heat and Humidity) Studies: Samples were tested under different temperature and humidity conditions in the environmental chamber.
In alignment with our mission to improve the overall health, safety, and well-being of people everywhere, our team worked to accelerate the study process. Through strong collaboration with project partners, we designed and modified our studies to meet customer needs, while maintaining effective communication throughout. Our commitment to this effort was key to its success and resulted in one of the fastest FDA marketing authorizations through De Novo request and CLIA Waiver application.
Because many of our laboratories are designed to be modular, we were able to install the customer’s equipment to enable an efficient and effective testing process. The laboratory that supported this testing, which is in our Diagnostics Center of Excellence, was quickly configured to ensure testing could be performed in parallel on eight different instruments.
Throughout testing, we sent LoD and flex testing data to the client and then to the FDA, further accelerating the process. It was through this collaborative effort and ongoing communication that prompt approval of the device was possible, enabling market access.
Read more about the Cepheid diagnostic at “Cepheid Receives FDA Authorization with CLIA Waiver for Xpert® HCV.”
This project has been funded in part with Federal funds from the National Institute of Biomedical Imaging and Bioengineering (NIBIB), National Institutes of Health, Department of Health and Human Services, under Contract No. 75N92022D00013.
