A novel multiplex PCR for the simultaneous detection of Salmonella enterica and Shigella species

• Published in: Brazilian journal of microbiology Vol. 45; no. 2; pp. 667 - 676
• Main Authors: Radhika, M, Saugata, Majumder, Murali, H S, Batra, H V
• Format: Journal Article
• Language: English; Portuguese
• Published: Brazil Springer Nature B.V 01.04.2014; Sociedade Brasileira de Microbiologia
• Subjects: Animals; Bacteriological Techniques - methods; Cell culture; Chickens; Environmental Microbiology; Food Microbiology; Gastrointestinal diseases; Genes; Genetics and Molecular Microbiology; Humans; MICROBIOLOGY; Molecular Diagnostic Techniques - methods; Multiplex Polymerase Chain Reaction - methods; Salmonella; Salmonella enterica - isolation & purification; Salmonella Infections - microbiology; Salmonella Infections, Animal - microbiology; Sensitivity and Specificity; Shigella - isolation & purification; multiplex PCR; Salmonella; spiking; Shigella
• ISSN: 1517-8382; 1678-4405; 1678-4405
• DOI: 10.1590/S1517-83822014005000041

Salmonella enterica and Shigella species are commonly associated with food and water borne infections leading to gastrointestinal diseases. The present work was undertaken to develop a sensitive and reliable PCR based detection system for simultaneous detection of Salmonella enterica and Shigella at species level. For this the conserved regions of specific genes namely ipaH1, ipaH, wbgZ, wzy and invA were targeted for detection of Shigella genus, S. flexneri, S. sonnei, S. boydii and Salmonella enterica respectively along with an internal amplification control (IAC). The results showed that twenty Salmonella and eleven Shigella spp., were accurately identified by the assay without showing non-specificity against closely related other Enterobacteriaceae organisms and also against other pathogens. Further evaluation of multiplex PCR was undertaken on 50 natural samples of chicken, eggs and poultry litter and results compared with conventional culture isolation and identification procedure. The multiplex PCR identified the presence of Salmonella and Shigella strains with a short pre-enrichment step of 5 h in peptone water and the same samples were processed by conventional procedures for comparison. Therefore, this reported multiplex PCR can serve as an alternative to the tedious time-consuming procedure of culture and identification in food safety laboratories.