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

Abstract

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.

A review on detection methods used for foodborne pathogens

“…The detection limits can become better with time that the DNA amplicon detection limit can be as low as femtograms (10 -15 g) 9 . This can be an alternative to tedious time consuming procedure of culturing and identification of pathogens in food safety laboratories 48 . However, with the advancement in PCR technology, the method has not remained cost-effective although low detection limit will remain the major criteria.…”

DNA sequencing, genomes and genetic markers of microbes on fruits and vegetables

“…ID 496 113 1221_SBOY 4.84 50.7 5125 4745 ipaH, 16S rRNA (Warren et al, 2006); invC (Ojha et al, 2013); wzy (Radhika et al, 2014); genome marker (Sahl et al, 2015;Kim et al, 2017a (Warren et al, 2006); IS1 (Hsu et al, 2007); she PAI (Farfan et al, 2010); invC, wbgZ (Ojha et al, 2013); IpaBCD (Farshad et al, 2015); genome marker (Sahl et al, 2015;Kim et al, 2017a) S. dysenteriae ID 415 67 Sd197 4.56 50.9 4834 4294 ial, virA, 16S rRNA (Warren et al, 2006); invC, rfpB (Ojha et al, 2013); genome marker (Sahl et al, 2015;Kim et al, 2017a) Campylobacter C. jejuni ID149 1615 NCTC 11168 1.64 30.5 1668 1572 pDT1720 (Ng et al, 1997); CC2, CJ2 (Sails et al, 2001); ciaB, pldA, CDT (Ghorbanalizadgan et al, 2014); GTPase gene, hip, 16S rRNA, rrs, cdaF, porA, 16S-23S ITS, Hyp, cjaA, ceuE, hipO, mapA, ceuA, askD, glyA, lpxA, ccoN, ORF-C sequence, rpoB, oxidoreductase gene, cdtA, pepT (Frasao et al, 2017); flaA, cadF, racR, dnaJ, cdtB, and cdtC (Oh et al, 2017); gyrA (Sierra-Arguello et al, 2018); ask, cdt (Kabir et al, 2019) C. coli ID1145 928 Aerotolerant OR12…”

Đánh giá khả năng nhiêm Listeria monocytogenes, staphy-lococcus aureus và salmonella SPP. Trong các mẫu sữa thu thập tại Gia Lâm và Ba Vì, Hà Nội đầu năm 2019

“…[14] với độ nhạy của phản ứng PCR nhận biết vi khuẩn Salmonella spp. là 0,3 pg/µL, trong khi với nhóm Radhika là 5 pg/µL [15] hay Kumar et al là 3 pg/µL [16]. Như vậy, trong trường hợp nhận biết Salmonella spp.…”

“…kết quả của chúng tôi nằm ở mức trung bình so với các công bố của các nhóm nghiên cứu này. So với hai nhóm sau thì thậm chí kết quả của chúng tôi tốt hơn [15,16].…”