Development and Standardization of the SHERLOCK (Specific High-Sensitivity Enzymatic Reporter Unlocking) Platform Combined with Lateral Flow Assay for Rapid Diagnosis of Infectious Laryngotracheitis and Avian Infectious Bronchitis

• Grant number: 25/10812-0
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: July 01, 2025
• End date: June 30, 2028
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Preventive Veterinary Medicine
• Principal Investigator: João Pessoa Araújo Junior
• Grantee: Amanda Haisi
• Host Institution: Instituto de Biotecnologia (IBTEC). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Associated research grant: 21/11968-2 - Center of Technological Plataform on Animal Health, AP.CCD

Abstract

Respiratory diseases in poultry represent one of the main causes of economic losses in the industrial poultry sector, especially those associated with viral infections that are subject to mandatory notification, such as infectious laryngotracheitis (ILT) and avian infectious bronchitis (IB). Currently, Brazilian poultry production has a significant impact on chicken meat exports, in addition to standing out in the production and export of eggs. The state of São Paulo ranks second nationally in the export of fertile eggs and third in the export of commercial table eggs, highlighting the importance of projects aimed at poultry health, especially the development of diagnostic tests that are rapid, sensitive, specific, and easy to apply, directly contributing to the prevention and control of diseases that affect the production chain. Regarding the methodologies used for the diagnosis of both viruses, molecular techniques for nucleic acid detection, such as polymerase chain reaction (PCR), with or without reverse transcription (qPCR and RT-qPCR), have been widely used. However, more recent technologies based on CRISPR/Cas systems are under constant development and offer greater analytical sensitivity. Rapid, sensitive, and specific detection methods are essential for diagnostics, particularly when combined with lateral flow detection systems, which enable quick visualization and require minimal laboratory infrastructure, thereby allowing faster and more efficient diagnosis of viral diseases. In this context, this project aims to develop a detection system based on the SHERLOCK platform (Specific High-Sensitivity Enzymatic Reporter Unlocking), combined with lateral flow readout, for the detection of infectious laryngotracheitis virus (ILTV) and avian infectious bronchitis virus (IBV), offering a practical and user-friendly device that requires only basic laboratory equipment for its operation. For this purpose, reagents will need to be imported and produced. The developed detection systems will be compared with diagnostic assays already validated by the World Organisation for Animal Health (WOAH) and applied to clinical samples. This project is expected to result in the development of a practical method for the detection of both viruses, with the potential to be integrated into routine poultry health monitoring. (AU)