title: Development of IgY based sandwich ELISA for the detection of staphylococcal enterotoxin G (SEG), an egc toxin.
abstract: Staphylococcal food poisoning (SFP) is a major foodborne illness caused by staphylococcal enterotoxins (SEs). It is a well known fact that foodborne outbreak investigations are solely characterized by commercially available immunoassay kits. However, these assays encompass only few enterotoxins such as SEA-SEE which are renowned as "classical" enterotoxins and unable to detect any other novel enterotoxins even though their involvement is predicted. In this context, the present study involved development of a sandwich ELISA immunoassay for the specific detection of "non-classical" enterotoxin G (SEG). The toxin belongs to enterotoxin gene cluster (egc) which comprises a bunch of five toxin genes that are known to co-express. Thus, the developed assay might indirectly speculate the presence of other toxins in the cluster. The efficiency of ELISA was compared with PCR analysis where all strains possessing seg were found positive for toxin production. Additionally, analogous to other studies which reported the co-occurrence of seg and sei, the PCR analysis accomplished in the study evinced the same. The sandwich format allowed sensitive detection with a detection limit of 1ng/mL. High specificity was achieved in presence of non-target protein as well as bacteria. Likewise, staphylococcal protein A (SpA) interference that is inevitably associated with immunoassays was eliminated by implementation of anti-SEG IgY in our study. Consequently, chicken IgY were used to capture target antigen in developed sandwich ELISA. Further, spiking studies and analysis on natural samples emphasized the robustness as well as applicability of developed method. Altogether, the established assay could be a reliable detection tool for the routine investigation of SEG as well as to predict other egc toxins in samples from food and clinical sources.
authors: Microbiology Division, Defence Food Research Laboratory, Siddarthanagar, Mysore, Karnataka 570011, India.
authors: Sowmya
authors: S
authors: Nagaraj
authors: Microbiology Division, Defence Food Research Laboratory, Siddarthanagar, Mysore, Karnataka 570011, India. Electronic address: shylajaramlal@gmail.com.
authors: Shylaja
authors: S
authors: Ramlal
authors: Microbiology Division, Defence Food Research Laboratory, Siddarthanagar, Mysore, Karnataka 570011, India.
authors: Joseph
authors: J
authors: Kingston
authors: Microbiology Division, Defence Food Research Laboratory, Siddarthanagar, Mysore, Karnataka 570011, India.
authors: Harsh Vardhan
authors: HV
authors: Batra
mesh_terms: D000818:Animals; D004768:Enterotoxins / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay; D005506:Food Contamination; D005516:Food Microbiology; D007136:Immunoglobulins / Q000032:analysis*; D005810:Multigene Family; D016133:Polymerase Chain Reaction; D011817:Rabbits; D015203:Reproducibility of Results; D013202:Staphylococcal Food Poisoning / Q000382:microbiology; D013211:Staphylococcus aureus / Q000235:genetics*; D018089:Superantigens / Q000302:isolation & purification*
chemical_list: D004768:Enterotoxins; C016512:IgY; D007136:Immunoglobulins; D018089:Superantigens; C419397:enterotoxin G, staphylococcal
keywords: IgY; SEG; Sandwich ELISA; SpA; Staphylococcal enterotoxins; egc