Quantitative diagnostic method to detect Gardnerella vaginalis by droplet digital PCR.

BACKGROUND

Nucleic Acid Amplification Tests (NAAT) remain one of the most reliable methods for pathogen identification. Given the high false-negative rates associated with traditional staining and microscopic examination, the time-consuming nature and low sensitivity of bacterial culture methods, as well as the inability of conventional NAAT to achieve absolute quantification.

METHODS

To achieve rapid and quantitative detection of Gardnerella vaginalis, we selected the 23S rRNA gene as the target for identification and developed a droplet digital PCR detection method.

RESULTS

The entire detection process can be completed within 92 min, demonstrating high efficiency. The sensitivity reached 4.4 pg/μL, and no positive droplets were detected in experiments involving eight negative control pathogens, confirming high specificity. Additionally, the ddPCR assay for Gardnerella vaginalis exhibited excellent repeatability, with a calculated coefficient of variation of 1 %.

CONCLUSION

The ddPCR detection technology demonstrates characteristics such as absolute quantification, high sensitivity, high specificity, and high reproducibility for Gardnerella vaginalis, showing promise as an excellent testing platform. This advancement could provide a more scientific basis for clinical diagnosis and treatment.