New method developed for target DNA sequence amplification, testing and analysis

A team of researchers from the Case Western Reserve University School of Medicine has developed a new method for target DNA sequence amplification, testing and analysis.

This new technique, or reaction, known as AMPLON (Amplifying DNA with Multiarm Priming and Looping Optimization of Nucleic Acid), offers an alternative to the previously accepted "gold-standard" Polymerase Chain Reaction (PCR) method, opening the opportunity for more applications in medical diagnosis.

The team's findings were recently published in the journal Advanced Materials.

"AMPLON has the potential to positively change the way molecular analysis and clinical diagnostics are performed," said Mohamed S. Draz, an assistant professor at the School of Medicine and the study's principal investigator, "from infectious-disease diagnostics to personalized medicine and environmental monitoring."

How it works

Researchers use such technology to compare the DNA of sick cells to that of healthy cells, allowing them to better understand the changes that occur as a disease progresses and how to treat it.

AMPLON provides several extensions along the DNA strand to simultaneously increase the speed and accuracy of DNA synthesis under constant temperature conditions.

Using this new simplified process eliminates the need to operate between high and low temperature extremes that can cause stress on materials. It also makes the amplification process more structured and accessible, especially in settings where precise temperature control is challenging.

Using the traditional PCR method, the DNA sample is heated so it can separate into two pieces of single-stranded DNA. Next, an enzyme builds two new strands of DNA, using the original strands as templates. The process is tedious, time consuming and expensive.

We've developed a new method of DNA amplification that does not require bulky lab-bound equipment but can be conducted in one step and in diverse settings. More significantly, our approach does not weaken enzymes like the PCR method."

Mohamed S. Draz, assistant professor, Case Western Reserve University School of Medicine

AMPLON's multiarmed DNA primer design can turn the shortcomings of enzymes into strengths to improve amplification efficiency and produce consistent results.

"We've been able to enhance amplification and reduce amplification time by 50%," Draz said. "Our approach has the potential to dramatically change the way nucleic acid amplification is performed, providing instead a portable, reliable and cost-effective solution for applications, ranging from point-of-care diagnostics to field-based research." said Draz.

Source:
Journal reference:

Doganay, M. T., et al. (2024). AMPLON: Amplifying DNA with Multi‐Arm Priming and Looping Optimization of Nucleic Acid. Advanced Materials. doi.org/10.1002/adma.202311634.

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