But some scientists at the Singapore Agency for Science, Technology and Research (A-STAR) had their ears on. ‘We heard from other channels in December that there is some kind of pneumonia in Wuhan,’ recalled Dr Sebastian Maurer-Stroh, Deputy Executive Director (Research) of the A-STAR Bioinformatics Institute (BII).
Haunted by memories of past epidemics, such as SARS in 2003, swine flu in 2009 and Zika in 2016, local researchers took action even before the virus reached local shores. With the advancement of sequencing technologies over the years, the allocation of the SARS-CoV-2 genome was published through Chinese researchers in mid-January, allowing scientists around the world to temporarily create their diagnostic kits.
By the first week of February, A-STAR researchers and their staff at Tan Tock Seng Hospital (TTSH) had already submitted a locally evolved COVID-19 diagnosis, called fortitude Kit, and sent it to various local hospitals, both. local and foreign. In contrast, in the 2003 SARS outbreak, diagnostic kits were manufactured for them in Singapore only a few months after the outbreak began. Here’s a look at the accelerated tour of the Fortitude kit from the bank to the header.
Detecting a stealth killer
The Fortitude kit is based on a strategy known as real-time transcription polymer (RT-PCR) chain reaction. The test procedure begins with the collection of nasal swabs or nasopharyngeals from Americans suspected of having COVID-19. Once transported to the lab, the viral RNA is removed from these swabs. This is where the strength kit comes into the equation.
Each kit comes with a one-step RT-PCR test, which can be used with all required reagents in the right quantities. These reagents come with opposite transcriptase, an enzyme that converts viral RNA into DNA, as well as short DNA sequences called primers that can stumble upon the unique genetic footprint of SARS-CoV-2. During RT-PCR, primers recognize and bind to changed viral DNA sequences of RNA. Repeated heating and cooling cycles cause Taq Polymerever to exponentially create the same copies of viral DNA until they stumble upon the machine.
The Fortitude kit takes only about 90 minutes to complete, however, the preparation steps are added to a few hours and the effects may take a day or more to be published, depending on the resources of the physical care providers.
However, RT-PCR is widely considered to be the popular gold to detect viruses like SARS-CoV-2. Unlike other diagnostics on the market, RT-PCR-based controls have specificity rates of more than 99%, meaning that other people who test positive have the disease. The sensitivity of RT-PCR controls is based on the level of the disease, which approaches 100% in the first week of the disease, but then adapts less delicate over time. Until now, Fortitude has become one of the most followed diagnostic kits on the market.
All on deck
The Fortitude kit was made imaginable by a tough team of leading scientists from Singapore, namely A-STAR scientists, Dr. Maurer-Stroh and Dr. Masafumi Inoue; and the executive director of the Diagnostic Development Center (DxD), Dr. Sidney Yee and Dr. Timothy Barkham of TTSH. Each expert brought his own skill set specialized in the opposite combat to COVID-19.
With his experience in computational biology, Maurer-Stroh tested the genetic sequences of the virus and its evolution over time, sharing its concepts in the public database of the Global Influenza Data Sharing Initiative (GISAID). Based on this data, Inoue, head of the diagnostics organization at A-STAR’s Experimental Drug Development Center (EDDC), began expanding the Prototype Fortitude Kit. The first agenda? Design primers that point to SARS-CoV-2.
“Ensuring quality primer designs is our purpose when creating diagnostic kits,” Inoue said. To do this, his team first had to identify a separate series to point out that was also shared through existing epidemic strains. This forced Inoue to compare and contrast several coronavirus genomes, employing colleagues such as Maurer-Stroh to refine the selection of regions to target using bioinformatics. In the end, they opted for a region that encodes an enzyme that catalyzes RNA replication from an RNA matrix.
“We chose this region as our number one target because we knew it had the least number of mutations in the SARS virus in 2003,” Inoue explained. By designing primers based on unique portions of the virus that are less prone to mutations, more circulating viral strains can be detected seamlessly through the kit.
Once the primers and prototype were in place, TTSH’s Barkham stepped in to refine the kit parameters. Your equipment suppresses the RT-PCR procedure elsewhere, running it at other concentrations or temperatures to identify ideal conditions. They also evaluated the kit on samples from genuine patients, which can involve molecules that can confuse control and give erroneous readings.
Then, the final touches of Fortaleza were added through the Yee team of DxD Hub, a national platform run through A-ccelerate, A-STAR’s marketing arm. “We optimized verification to make sure it works every single time,” he shared. “We’ve also evolved the entire production process, because it’s not just about combining elements into a kit. It’s also about many production instructions, quality protocols and quality assurance.”
Given the great effort required to expand a diagnosis, it is notable that A STAR and TTSH have been able to do so in less than a month. Its long sleepless nights have been driven by an absolute willingness to help Singapore cope with its biggest crisis to date. “This is just a small glimpse of the team involved in deploying a singles product, Fortitude Kit,” Yee said. “The entire R&D ecosystem in Singapore is needed and we are very lucky to be a component of it.”
Singapore for the world
In Singapore, the Fortitude kit has been commonly used in thirteen public and personal hospitals and laboratories since February. To date, the kit has also been implemented in more than 20 countries. While the DxD Hub produced the initial batch of kits, knowledge was transferred not exclusively to a handful of biotechnology corporations to expand and manufacture diagnostic tests.
From a weekly production of 100,000 checks at the start of the epidemic, the production movement trains higher production up to more than 4 times per week. “Fortitude is now offered in about 20 countries.” This is one of the most widely used RT-PCR control kits in the world,” Yee said. In addition to its apparent clinical advantage, Fortitude Kit has also helped spread the Singapore and A-STAR logo symbol worldwide, he added.
However, the paintings continue. “RNA viruses are changing,” Maurer-Stroh explains. Most mutations are minuscule changes, however, following the evolution of the virus closely allows A-STAR to react temporarily and modify the Strength if necessary.