Abstract: TH-PO0893
Development of a Quantitative Polymerase Chain Reaction (qPCR)-Based Pathogen-Screening Protocol for Fetal Porcine Tissue Intended for Xenotransplantation
Session Information
- Transplantation: Basic Research
November 06, 2025 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Transplantation
- 2101 Transplantation: Basic
Authors
- Ohashi, Hinari, Division of Nephrology and Hypertension, Department of Internal Medicine Jikei, University School of Medicine, Minato, Tokyo, Japan
- Yamanaka, Shuichiro, Division of Nephrology and Hypertension, Department of Internal Medicine Jikei, University School of Medicine, Minato, Tokyo, Japan
- Matsui, Kenji, Division of Nephrology and Hypertension, Department of Internal Medicine Jikei, University School of Medicine, Minato, Tokyo, Japan
- Matsumoto, Kei, Division of Nephrology and Hypertension, Department of Internal Medicine Jikei, University School of Medicine, Minato, Tokyo, Japan
- Sakurai, Tatsuya, Laboratory Animal Facilities, Jikei University School of Medicine, Minat, Tokyo, Japan
- Yokoo, Takashi, Division of Nephrology and Hypertension, Department of Internal Medicine Jikei, University School of Medicine, Minato, Tokyo, Japan
Background
Fetal porcine kidneys have emerged as promising donor organs for xenotransplantation because they do not trigger vascular hyperacute rejection. Although fetal organs develop in the intrauterine environment and are less exposed to external pathogens, several microorganisms capable of transplacental transmission have been identified. To ensure the microbiological safety of xenotransplantation, reliable pathogen screening methods are essential. Although PCR-based techniques are widely used for pathogen screening in clinical and research settings, there is currently no standardized protocol specifically designed for fetal tissues. This study aims to establish a robust qPCR-based screening protocol for fetal porcine tissues, addressing a critical gap and supporting their safe application in xenotransplantation.
Methods
1. Target pathogens were selected in accordance with the xenotransplantation guideline issued by the Japanese Ministry of Health, Labour and Welfare (MHLW) and based on relevant literature and epidemiological data.
2. Fluorescent probe-based qPCR assays were developed for the target pathogens using either commercially available or customized qPCR kits.
3. The performance and analytical sensitivity of each qPCR assay were assessed by evaluating the limit of detection (LOD), through serial dilutions of positive control nucleic acids.
In all experiments, aseptically collected fetal tissue, excluding the kidneys and bladder, was homogenized in its entirety, and an aliquot of the homogenate was used for analysis.
Results
1. Twenty viruses and 2 parasites known to cause transplacental infections in pigs and reported in Japan were selected as target pathogens. All bacteria and fungi listed in the MHLW guideline were also included.
2. qPCR assays were successfully developed for 20 viruses, one parasite (excluding one due to the lack of available genome information), and a broad range of bacteria and fungi.
3. The developed qPCR assays demonstrated promising sensitivity, with a LOD below 100 copies.
Conclusion
We have established a list of pathogens relevant to xenotransplantation using fetal porcine tissue and developed qPCR assays for their detection. These assays offer a valuable foundation for establishing pathogen screening panels tailored to xenotransplantation involving fetal porcine organs.