Abstract
In many countries, Salmonella is the leading cause of food-borne outbreaks and infections. A multiplex PCR (mPCR) for the detection of genus Salmonella and serogroups A, B, C1, D and capsular (Vi) producing strains from swabs of stool samples was developed. In the mPCR, primers for invasion (invA), O (prt, tyv, rfbj, wzxC1) and Viantigen genes (Vi) and internal amplification control primers were used. The results showed that all tested Salmonella serotypes were accurately identified by the assay, without nonspecific amplification except Salm. derby and Salm. saint Paul. Representative serogroups were used to artificially inoculate stool samples. The different serogroups were detected by mPCR after overnight pre-enrichment of stool swab in buffered peptone water with a detection limit of Salmonella cell suspension of 4 cfu/ g stool. The developed mPCR assay provides specific detection of genus Salmonella and serogroups A, B, C1, D and Vi positive strains directly from stool swabs. The developed method for Salmonellaserogroup identification is rapid, easy and less subjective methods. This could be of great use by any facility that lacks the expensive typing sera and expertise needed for conventional serotyping. Key words: Salmonella, multiplex PCR, stool, molecular serotyping, somatic antigen.
Highlights
Salmonella enterica is one of the major bacterial agents that cause foodborne infections in humans all over the world (Herikstad et al, 2002)
The results showed that all tested Salmonella serotypes were accurately identified by the assay, without nonspecific amplification except Salm. derby and Salm. saint Paul
A multiplex PCR assay was developed for the rapid detection of genus Salmonella and the most prevelant serogroups in Egypt, as isolated by Egyptian Central Health Laboratories (ECHL), namely A, B, C1, D and Vi producing strains from stool
Summary
Salmonella enterica is one of the major bacterial agents that cause foodborne infections in humans all over the world (Herikstad et al, 2002). The Salmonella serotype is determined on the basis of somatic (O) antigen which determines the group and flagellar (H) antigen which determines the serotype (Popoff 2001). O antigen is a specific polysaccharide; the genes for O-antigen are grouped together on the chromosome in a gene cluster called rfb (Bastin and Reeves, 1995). The basis of the variation in O antigen structure is represented by the different types of sugar present, the arrangement of sugars, the addition of branch sugars, and modifying side groups; such variation is used to design serogroup specific probes (Luk et al, 1993). The viaB consists of the structural genes specific for Vi
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