Detection ofin Microbiological Culture: A Comparative Analysis of PCR Primer Sets.

Abstract

Glanders is a highly contagious and often fatal zoonotic disease of equids caused by, a pathogen of significant concern due to its potential for bioterrorism. In Brazil, glanders remains endemic, particularly among working equids in the Northeast region. Diagnostic confirmation typically involves serology, culture, and polymerase chain reaction (PCR), although false-negative PCR results have been increasingly reported. This study aimed to evaluate the diagnostic performance and analytical sensitivity of four-specific PCR primer sets using samples from 30 seropositive equids. Microbiological cultures were obtained from various organs and swabs, followed by PCR targeting four genomic regions:-IS407A(a),-IS407A(b), Burk457, and Bm17. All animals were confirmed positive forvia culture, but PCR detection rates varied significantly across primer sets. The-IS407A(b) primer set showed the highest sensitivity, detecting 86% of samples, while the WOAH-recommended-IS407A(a) set had the lowest performance (13.4%). Analytical sensitivity assays confirmed that-IS407A(b) and Bm17 primers detected DNA concentrations as low as 0.007 ng, outperforming the others. These findings suggest that certain widely used primer sets may lack sufficient sensitivity for reliable detection of, especially in chronically infected animals with low bacterial loads. The study underscores the need for ongoing validation of molecular diagnostics to improve the detection and control of glanders in endemic regions.