New method enables faster, more accurate identification of pathogens, even in complex DNA sequences

A team of researchers at Case Western Reserve University School of Medicine has developed a new method for amplifying, testing and analyzing target DNA sequences.

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, paving the way 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, assistant professor in the School of Medicine and principal investigator of the study, “from diagnosing infectious diseases to personalized medicine and environmental surveillance.

How it works

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

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

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

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

“We have developed a new method for DNA amplification that does not require bulky laboratory equipment, but can be performed in a single step and in a variety of settings,” Draz said. “More importantly, our approach does not weaken enzymes like the PCR method.”

AMPLON's multi-arm DNA primer design can turn enzyme flaws into strengths to improve amplification efficiency and produce consistent results.

“We were able to improve amplification and reduce amplification time by 50 percent,” Draz said. “Our approach has the potential to radically change the way nucleic acid amplification is performed, instead providing a portable, reliable and cost-effective solution for applications ranging from point-of-care diagnostics to on-site research. ground.”