The Agency for Science, Technology and Research (A*STAR), and Tan Tock Seng Hospital (TTSH), Singapore, have successfully developed a test kit which can detect the presence of the SARS-CoV-2
virus which causes COVID-19, quickly and with high accuracy.
Called Fortitude Kit 2.0, the test was co-developed with, and validated by TTSH. It is the first “ready-made” hospital lab diagnostic test kit that has received Singapore Health Sciences Authority’s (HSA) Provisional Authorisation for clinical use. Each kit comes complete with all the necessary reagents prepared in the right quantities, and quality-controlled tubes.
Since the SARS outbreak in 2003, lead scientists at A*STAR have advanced Singapore’s R&D capabilities to study infectious diseases, and worked closely with the clinical community to develop a robust diagnostic test in less than a month.
Dr Sebastian Maurer-Stroh, Deputy Executive Director (Research), A*STAR’s Bioinformatics Institute (BII) examined the genetic material of the SARS-CoV-2 and studied the virus evolution. His analysis was shared with the Global Initiative on Sharing all Influenza Data (GISAID), a platform which promotes the international sharing of all influenza virus sequences.
With the information, Dr Masafumi Inoue, Head of Diagnostics Group at EDDC, A*STAR, designed the primers to detect various gene targets of SARS-CoV-2, as well as developed the diagnostic prototype.
Associate Professor Dr Timothy Barkham, Senior Consultant Medical Microbiologist from the Department of Laboratory Medicine, TTSH, provided the clinical expertise and utility on the type of assays that is most useful, helped optimise test parameters, created positive control material, and evaluated the kit on clinical samples.
A*STAR worked with the National Public Health Laboratory at the National Centre for Infectious Diseases (NCID), Singapore, on the appropriate manufacturing standards of the tests. The Diagnostics Development (DxD) Hub, a national initiative led by A*STAR’s commercialisation arm, A*ccelerate, further supported the development and production of these tests.
Since February 2020, Fortitude Kit 2.0 has been implemented in six local public hospitals and one private hospital group in Singapore. 10,000 tests were also donated to China, and 3,000 tests were sent to Myanmar as part of Singapore’s efforts to curb the spread of the disease. More are currently being delivered to various countries across the globe. A*STAR is currently transferring the technology and know-how to companies with a non-exclusive license to scale-up and manufacture the diagnostic kits to fulfil local and international demand.
To complement the Fortitude Kit 2.0, A*STAR and TTSH are also currently working on a Multiplex Diagnostic Kit that can simultaneously differentiate between SARS-CoV-2 and influenza viruses, and assist healthcare professionals to make accurate diagnoses. Also in the pipeline is a rapid point-of-care-test, which could potentially expand the availability of testing in locations such as at immigration checkpoints.
THE TECHNICAL SPECIFICATIONS OF FORTITUDE KIT 2.0
- A 1-step RT-PCR test, Fortitude Kit 2.0 runs on the real-time Reverse Transcription-Polymerase Chain Reaction (RT-PCR) laboratory technique that detects SARS-CoV-2 virus genetic material.
- This is a duplex RT-PCR assay which simultaneously detects the SARS-CoV-2-RNA and the internal control in a single reaction. The test takes approximately 90 minutes to churn out real-time RT-PCR results.
- The analytical limit of detection (LoD) of the A-STAR Fortitude Kit 2.0 was determined to be 25 copies of positive template per reaction. This means that if there were at least 25 copies of the SARS-CoV-2 virus nucleic acid material in a single test, the assay would be able to detect the virus with 95% confidence. It also has no cross-reactivity with other closely-related respiratory viruses including SARS-CoV, MERS-CoV and influenza.
- In terms of clinical sensitivity, Fortitude Kit 2.0 has been able to detect 100 per cent of the positive samples tested at TTSH, with no false positive and false negative reported; while other similar kits in the market have detected with less than 100 per cent clinical sensitivity.
- Fortitude Kit 2.0 targets the portion of the virus genome that is stable and less prone to mutation, therefore enabling robust detection of the virus. To date, no mutation has been detected in the targeted area.
- That said, there is also continuous effort to monitor the target region of the virus genome. This ensures that the assay performance of Fortitude Kit 2.0 is not affected by a potential mismatch between the primer probes and the targeted gene sequences, that could lead to compromised results.