A highly sensitive method for the detection of recombinant PERV-A/C env RNA using next generation sequencing...

Area of Science:

• Xenotransplantation research
• Virology
• Biotechnology

Background:

• Xenogenic cell-based therapies, like porcine islet cells for diabetes, face PERV transmission risks.
• Four PERV subgroups exist (PERV-A, -B, -C, PERV-A/C), with PERV-A/C showing high prevalence and potential viremia increase in diseased pigs.
• Monitoring PERV-A/C emergence post-transplantation is critical for xenotransplantation safety.

Purpose of the Study:

• To develop a highly sensitive detection method for PERV-A/C.
• To assess the utility of next-generation sequencing (NGS) for PERV-A/C detection.

Main Methods:

• Developed a sensitive method using next-generation sequencing (NGS) technologies.
• Utilized RT-PCR to amplify PERV-C spiked with varying PERV-A/C doses.
• Analyzed amplicons via NGS.

Main Results:

• NGS successfully detected PERV-A/C at 1% abundance with 100% true positive rate.
• NGS also detected PERV-A/C at 0.1% abundance with a 57% true positive rate.
• The method demonstrates high sensitivity for PERV-A/C detection.

Conclusions:

• The developed NGS method is highly sensitive and effective for detecting PERV-A/C.
• This technique can be valuable for rapid monitoring of PERV-A/C emergence after xenotransplantation.
• Ensures the safety and efficacy of porcine-derived cell therapies.