Utilizing HRPzyme, a cost-effective Vibrio parahaemolyticus detection method

Vibrio parahaemolyticus is a prominent infectious bacterium responsible for causing widespread cases of acute gastroenteritis in humans globally. In this regard, Colorimetric detection can be essentially used as a sensitive, rapid, and cost-effective detection method. In our research, we have developed a PCR-based detection platform integrated with HRPzyme and utilizing DNAzyme as a signaling probe which mimics peroxidase activity. The colorimetric signal is detectable at concentrations as low as 101 cfu mL-1 when measured with a spectrophotometer and at 103 cfu mL-1 through visual inspection. Additionally, extending the polyadenine length to 10 nucleotides resulted in a significant reduction in the background signaling of HRPzyme activity, yielding a relative intensity of 3.07 +/- 0.23 arbitrary units (a.u.). Notably, even after a 120-min incubation period, there were no further changes observed in the colorimetric signal in positive samples, maintaining a consistent relative intensity of OD 410 = 0.55 +/- 0.08.

Automated summary

We have developed a PCR-based detection platform integrated with HRPzyme and DNAzyme as a signaling probe to detect Vibrio parahaemolyticus. The colorimetric signal can detect bacterial colonies at concentrations as low as 101 cfu/mL. Extending the polyadenine length reduces background signaling and improves detection accuracy.