IISc develops tech as RT-PCR alternative – ET HealthWorld

Bengaluru: A team of Indian Institute of Science (IISc) researchers has reported a new technique for detection of viruses that could help overcome the limitations and challenges of real-time Reverse Transcription-Polymerase Chain Reaction (RT-PCR).

RT-PCR is the standard test used to detect viruses such as Ebola and SARS-CoV-2. A throat swab or a blood sample may have only a few viral DNA or RNA molecules, making it hard to detect the virus.

“Therefore, the number of DNA or RNA molecules is amplified using PCR. This newly generated DNA is tagged by fluorescent molecules and monitored in real time to estimate the level of DNA (or viral RNA) in the sample. This technique also uses a thermal cycler which regulates temperature because different temperatures are required in each step of multiplying DNA or RNA molecules. However, the use of thermal cyclers and real-time monitoring limits the use of RT-PCR in resource-starved areas,” according to IISc.

The new method

To overcome this limitation, Priyanka Valloy and Rahul Roy from the department of chemical engineering, IISc, used an alternative method called ‘Recombinase Polymerase Amplification’ (RPA) in which all reactions take place at close to room temperature. “Developing an RPA kinetics mathematical model that estimates the amount of the initial DNA based on the amount of genetic material present at the end point, they developed a new method called ‘quantitative endpoint RPA (qeRPA)’ that eliminates the need for real-time monitoring,” IISc added.

The researchers then used qeRPA to determine the initial number of DNA copies in blood samples from dengue patients. “Results from qeRPA were consistent with those obtained from qRT-PCR while being faster and easier to implement. The team hopes this method could be used to detect nucleic acids like DNA or RNA at diagnostic centres in resource-limited areas like remote villages,” IISc said.

  • Published On Apr 5, 2023 at 02:35 PM IST

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