Standardization of preparation, extraction, and amplification of several types of samples to detect clinically important pathogens for Xenotransplantation realization

• Grant number: 25/03404-2
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: May 01, 2025
• End date: December 31, 2025
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Silvano Mario Attilio Raia
• Grantee: Mayumi dos Santos Hayami
• Host Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 21/11872-5 - Science for Development Center in Xenotransplantation, AP.CCD

Abstract

The scientific community asserts that the absence of Specific Pathogen-Free (SPF) and Designated Pathogen-Free (DPF) pathogens in xenotransplantation candidate animals is paramount. The real-time polymerase chain reaction (qPCR) is one of the most efficacious methodologies for detecting these pathogens, mainly due to its high sensitivity (1 copy/mL). The selection of samples to be evaluated is important for the effectiveness of pathogen detection, particularly in instances where latency may be present. Consequently, the combination of sample type, extraction kit, primers, probes, and master mix is an outstanding factor to obtain reliable results, thereby mitigating the risk of false negatives that could jeopardize the recipient's health and the viability of the transplanted organ.