Its new technique may allow for wider adoption of COVID-19 diagnostic tests in university or study laboratories, and enable detection and study, i.e. in countries and regions with limited laboratory capacity. The test, which can be performed with laptops, can also be deployed on the network as a discovery tool.
Currently, the most delicate approach to testing COVID-19 is to use a lab strategy called polymerase chain reaction (PCR), in which a device amplifies viral genetics by copying it over and over again so that any indication of the SARS-CoV-2 virus can be detected.
A large bottleneck in pattern testing is RNA purification (separation of RNA from other parts of the patient pattern), a difficult procedure that requires chemicals that are now rare in the world. Their arrangements must be carried out through a highly qualified technical staff and may take a few hours. Currently, automated pattern preparation appliances cost thousands of dollars and require specialized laboratories.
The approach developed through NTU LKCMedicine combines several of these steps and allows direct testing on the uncooked patient pattern, reducing processing time from pattern to result and eliminating the need for RNA purification chemicals.
Details of the novelty were published in the clinical journal Genes in June.
Mr. Wee Soon Keong, Ph.D. NTU candidate LKCMedicine and the first author of the paper said: “While polymerase chain reaction (PCR) is a venerable generation that has proven to be addicted to work for biological research, it has some drawbacks when used outdoors in the lab. The procedure is tedious and time-consuming. Our rapid coVID-19 control comes to a single-tube reaction that reduces handling time and biosecurity threat to lab staff, as well as the likelihood of a motion contamination pattern.”
In addition to the detection of COVID-19, it developed through the LKCMedicine NTU team can also be used to trip over other viruses and bacteria, adding the dengue virus, which is about to affect Singapore as the country prepares for one of the worst dengue. Fever. epidemics amid the coronavirus pandemic.
The head of the study team, associate professor Eric Yap, who also runs the Microbial Genomics Laboratory, said: “We have shown in the past that this approach also works for dengue virus. When used directly in a raw blood pattern containing the dengue virus, the control resulted in 28 minutes. As Singapore is fighting the double dengue epidemic and COVID-19, either with similar early symptoms, our control can help differentiate the two infectious diseases.”
Professor James Best, dean of NTU LKCMedicine, said: “As Singapore continues its proactive verification to detect, isolate and engage the imaginable spread of coronavirus, immediate portable detection equipment, such as the one developed through Assoc Prof Yap and its team, can simply enter the community control sites, allowing the immediate identity of inflamed patients, and immediate action to prevent transmission.”
From lab tests to tests
As a general rule, in PCR controls, it will be necessary to remove a genetic cloth in a swab pattern taken from a patient to remove any substance from the pattern that prevents the functioning of PCR control. An example of an inhibitor of respiratory patterns is mucin (a major component of mucus).
The control, designed through the LKCMedicine NTU team, which includes the principal investigator, Dr. Sivalingam Paramalingam Suppiah, uses the “direct PCR” method, eliminating the need for RNA purification, a long and costly step. Instead, they added inhibitor-resistant enzymes and reagents that target compounds that block RNA amplification, such as mucin, a major component of mucus. These enzymes and reagents, which are commercially available, have superior resistance to those compounds that differently inhibit PCR, making control inaccurate.
The biochemical aggregate of raw pattern and inhibitor-resistant enzymes and reagents is placed in a single tube, which is inserted into a laboratory thermocycler, a device used to magnify genetics in PCR. After 36 minutes, the effects are shown with confidence if there is an indication of COVID-19.
“By omitting the level of RNA extraction with our direct PCR method, we are seeing savings in nucleic acid extraction kits and avoiding the challenge of reagent scarcity when the lab intensifies and demands increases globally,” Dr. Sivalingam said.
The team also verified this approach on a portable thermocycler, which can be deployed in endemic and low-resource environments, indicating the option to schedule this verification on network fitness care services through number one fitness workers.
Associate Professor Yap said: “We are now looking to implement such direct PCR strategies, developed through ourselves and others, for the diagnosis of regimens. We want to discover the app and the genuine benefits in a genuine context, and perceive if there is trade. Off. When a bottleneck is eliminated, other demanding situations may arise, such as quality errors or manual cutting errors.”
The team will now use this approach for COVID-19 testing at LKCMedicine’s NTU Clinical Diagnostic Laboratory, led by Professor Assoc Yap.