Differentiation of non-aureus staphylococci species isolated from bovine mastitis by PCR-RFLP of groEL and gap genes in comparison to MALDI-TOF mass spectrometry. (December 2020)

• Record Type: Journal Article
• Authors: Conesa, Agustín; Dieser, Silvana; Barberis, Claudia; Bonetto, Cesar; Lasagno, Mirta; Vay, Carlos; Odierno, Liliana; Porporatto, Carina; Raspanti, Claudia
• Is Part Of: Microbial pathogenesis. Volume 149(2020)
• Publication Date: 2020-12
• Subjects: Species identification -- Non-aureus staphylococci -- groEL PCR-RFLP -- gap PCR-RFLP -- MALDI-TOF MS; Pathogenic microorganisms -- Periodicals; Pathology, Molecular -- Periodicals; Communicable Diseases -- microbiology -- Periodicals; Communicable Diseases -- parasitology -- Periodicals; Micro-organismes pathogènes -- Périodiques; Pathologie moléculaire -- Périodiques; Electronic journals; 616.9041
• DOI: 10.1016/j.micpath.2020.104489
• Languages: English
• ISSNs: 0882-4010

Abstract

Abstract: Intramammary infections (IMI) cause serious economic losses for farmers and the dairy industry. Cases of subclinical mastitis are commonly the result of infection by minor pathogens such as non- aureus staphylococci (NAS), so their correct identification is important for appropriate therapeutic intervention and management. The aim of this study was to assess the reliability of PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) of the groE L and gap genes to discriminate between bovine-associated NAS species, using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) as the reference method. MALDI-TOF MS was able to correctly identify 112 NAS isolates from bovine IMI at species level out of a total of 115 (97.4%). These results were considered definitive and thus compared with those from the PCR-RFLP analyses. Only 50% (56/112) of the samples classified through groEL PCR-RFLP matched the molecular identity determined by MALDI-TOF MS, whereas coincidence rose to 96.4% (108/112) when comparing results from gap PCR-RFLP and the spectral analysis. This study demonstrates that gap PCR-RFLP is a useful and reliable tool for the identification of NAS species isolated from bovine mastitis. Highlights: groEL and gap PCR-RFLP profiles for non- aureus staphylococci identification. The performance of two genotypic methods was comparatively evaluated. MALDI-TOF MS was used as a gold standard to resolve discrepancies. gap PCR-RFLP can better identify non- aureus staphylococci than groEL PCR-RFLP.