Salmonella Spp. and Shigella Spp. detection via multiplex real-time PCR and discrimination via MALDI-TOF MS in different animal raw milk samples

• Published in: Nigerian journal of clinical practice Vol. 22; no. 8; pp. 1083 - 1090
• Main Authors: Demirci, M, Yigin, A, Altun, S, Uysal, H, Saribas, S, Kocazeybek, B
• Format: Journal Article
• Language: English
• Published: India Wolters Kluwer India Pvt. Ltd 01-08-2019; Medknow Publications and Media Pvt. Ltd
• Subjects: Animals; Cattle; Dairy products industry; DNA; DNA, Bacterial - analysis; Epidemiology; Equidae; Female; Food Contamination - analysis; Food poisoning; Genetic aspects; Goats; Health aspects; Humans; Identification and classification; Ionization; Mass spectrometry; Milk; Milk - microbiology; Milk contamination; Polymerase Chain Reaction - methods; Risk factors; Salmonella; Salmonella - classification; Salmonella - genetics; Salmonella - isolation & purification; Sensitivity and Specificity; Sheep; Shigella; Shigella - classification; Shigella - genetics; Shigella - isolation & purification; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; Spectroscopy; Tetracyclines; Turkey; MALDI-TOF MS; Shigella spp; qPCR; Salmonella spp; subspecies discrimination
• ISSN: 1119-3077
• DOI: 10.4103/njcp.njcp_596_18

Aims: The aim of this study was to provide epidemiological data about the presence of Salmonella spp. and Shigella spp. in raw milk samples collected from different animals. Methods: A total of 231 raw milk samples from 48 cows, 65 goats, 65 sheep, and 53 donkeys were studied. The ISO 6579:2002 and ISO 21567:2004 methods, antimicrobial susceptibility tests, and serotyping were performed. Species and subspecies discriminations were made via matrix-assisted laser desorption/ionization-time of flight mass spectrometry. After DNA isolation from all samples, Salmonella spp. and Shigella spp. were detected using real-time polymerase chain reaction (PCR) kits. Results: Five samples (2.16%) showed positivity out of 231 raw milk samples for Salmonella spp., and 2 (0.87%) samples were detected to be positive by multiplex real-time PCR design. Conclusion: We found that raw milk samples were not free of Salmonella spp. and Shigella spp. and need to be tested routinely to avoid public health problems. Rapid and reliable real-time PCR method can be developed and used for this purposes instead of slow bacterial culture processes.