Advancement of the Isothermal Loop-Mediated Amplification Technique for fosA gene Detection in Bacterial Isolates from Urinary Tract Infections.

• Grant number: 23/03267-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: February 01, 2024
• End date: August 31, 2024
• Field of knowledge: Health Sciences - Collective Health - Public Health
• Principal Investigator: Inneke Marie van der Heijden Natário
• Grantee: Letícia Silva Figueiredo
• Host Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

The loop-mediated isothermal amplification technique (LAMP) is a cost-effective and field deployable molecular biology method that can be used for detecting a wide range of targets, including resistance genes such as carbapenem and fosfomycin resistance genes in bacterial isolates from urinary tract infections. LAMP interpretation can be done colorimetrically, offering the advantage of nearly instantaneous results that are applicable on a large scale and at a low cost. Fosfomycin is a single-dose medication indicated for the short-term treatment of uncomplicated bacterial infections of the lower urinary tract, and its resistance is still not well-studied. Urinary tract infections (UTIs) are one of the most prevalent infections across all age groups, with both community and healthcare-associated origins. The inappropriate use of broad-spectrum antibiotics for UTI treatment has been the main factor in the development of antibiotic resistance, becoming a global public health issue. This study aims to validate the primer for the fosfomycin resistance gene (fosA) for the development of LAMP technique with colorimetric detection. It also aims to evaluate the performance of LAMP by determining sensitivity and specificity in bacterial isolates from UTIs resistant to fosfomycin. The ultimate goal is to conduct the LAMP reaction in loco, directly on urine samples at the Basic Health Units (UBS), to facilitate diagnosis through bacterial counting and antibiotic susceptibility testing, thereby providing prompt treatment for these infections.