Clinical researchers at Nanyang University of Technology, Lee Kong Chian Medical School (LKCMedicine) in Singapore (NTU Singapore) have demonstrated a way to improve the speed, recovery time and load of COVID-19 laboratory tests.
The progressive verification approach provides effects in 36 minutes, a quarter of the time required by existing verifications.
Its new technique may allow for wider adoption of COVID-19 diagnostic tests in university or study laboratories, and allow detection and study, mainly 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 devices cost thousands of dollars and require laboratories.
The approach developed through NTU LKCMedicine combines several of these steps and allows direct testing of the patient’s raw pattern, reducing healing 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.
Although polymerase chain reaction (PCR) is a venerable generation that has proven to be a workhorse for biological research, it has some drawbacks when used outdoors in the lab. The procedure is tedious and time-consuming. Our immediate COVID-19 control reaches a single-tube reaction that reduces handling time and biosecurity threat to laboratory staff, as well as the likelihood of a pattern of motion contamination. “
In addition to the detection of COVID-19, it developed through the NTU LKCMedicine 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 cases of 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 with the dengue virus, verification resulted in 28 minutes.”
“While Singapore is battling the double dengue epidemic and COVID-19, either with similar early symptoms, our control can differentiate the two infectious diseases.”
Professor James Best, dean of NTU LKCMedicine, said: “As Singapore continues its proactive verification to detect, isolate and involve the imaginable spread of coronavirus, immediate portable detection equipment, such as the one developed through Assoc Prof Yap and his team, they can simply enter the community control sites, allowing the immediate identity of inflamed patients, and immediate action to prevent transmission.”
As a general rule, in PCR checks, it will be necessary to remove a genetic cloth in a swab pattern taken from a patient to remove all ingredients from the pattern that prevent the PCR test from working. An example of an inhibitor of respiratory patterns is mucin (a major component of mucus).
The test, designed through the LKC NTU Medicine team, which includes the principal investigator, Dr. Sivalingam Paramalingam Suppiah, uses the “direct PCR” method, eliminating the long and costly need for RNA purification.
Instead, they added enzymes and reagents resistant to inhibitory 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: “Now we are going to implement these direct PCR strategies, developed through ourselves and others, for the diagnosis of regimens.”
The team will now use this approach for COVID-19 testing at LKCMedicine’s NTU Clinical Diagnostic Laboratory, led by Professor Assoc Yap.
“Our purpose is to expand ultra-fast automated tests that produce effects in minutes and can be performed through health personnel in the clinic accurately and sensitively to those of specialized laboratories.
“This will allow us to take PCR testing from traditional labs closest to the point of service and in low-resource environments that need them most,” he said.
Nanyang University of Technology
Wee, S.K. et al. (2020) Quick verification of direct amplification of nucleic acid without RNA extraction for SARS-CoV-2, a portable PCR thermocycler. Genes. doi.org/10.3390/genes11060664.
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