Emerging wound-healing injectable polydeoxyribonucleotide: potential as a prohibited doping method and its simple...

Area of Science:

• Biotechnology
• Molecular Biology
• Forensic Science

Background:

• Polydeoxyribonucleotide (PDRN), derived from chum salmon DNA, promotes wound healing via growth factor upregulation.
• PDRN's regenerative properties raise concerns for potential athletic performance enhancement, constituting a form of doping.

Purpose of the Study:

• To develop a sensitive and specific method for detecting Polydeoxyribonucleotide (PDRN) misuse in sports.
• To address the World Anti-Doping Agency's (WADA) concerns regarding PDRN as a prohibited performance-enhancing substance.

Main Methods:

• Utilized the CRISPR-Cas12a system targeting conserved 12S and 16S rDNA sequences specific to salmonids (Oncorhynchus keta).
• Optimized a direct PCR method for amplifying target sequences from human plasma and urine without prior DNA extraction.
• Employed Cas12a-mediated detection with a fluorescent signal indicating successful target recognition.

Main Results:

• Achieved high sensitivity, detecting as little as 0.8 pg (0.3 genome copies) of chum salmon DNA per 10 μL of biological sample.
• Detection was completed within 90 minutes, surpassing the sensitivity of many existing doping agent detection methods.
• Demonstrated specificity for salmonid DNA, crucial for accurate PDRN misuse identification.

Conclusions:

• The developed CRISPR-Cas12a method offers a highly sensitive and rapid approach for detecting PDRN misuse in sports.
• This assay provides a valuable tool for anti-doping agencies to uphold fair competition.
• The method's performance surpasses current standards, potentially aiding in the identification of novel doping strategies.