Comparison of Reverse Passive Latex Agglutination Test and Immunoblotting for Detection of Staphylococcal Enterotoxin a and B

Abstract

Staphylococcal foodborne diseases resulting from consumption of food contaminated with staphylococcal enterotoxins (SEs) produced by certain strains of Staphylococcus aureus are the second most common foodborne illnesses in the world. Analytical methods are essential for routine monitoring purposes and safeguard public health. Different methods for SE detection have been proposed although their use in a complex matrix is often limited by the presence of substances that interfere with tests. In this article reverse passive latex agglutination (RPLA) and immunoblotting methods based on specific antibodies and currently available for SE detection have been compared. Culture filtrates from enterotoxin S. aureus strains isolated from cheese samples were identified by SET‐RPLA. Then the culture filtrates identified as staphylococcal enterotoxin A and staphylococcal enterotoxin B by RPLA test were analyzed with immunoblotting. The results obtained suggest that either SET‐RPLA or immunoblotting may be applied to culture filtrates for the detection of SEs with good correspondence of results. Although SET‐RPLA represents a simple method for routine monitoring purposes, a positive result by a rapid method (RPLA) is only regarded as presumptive and must be confirmed by standard methods (Feng 1996), such as immunoblotting method.

Monoclonal Antibody-Based Sandwich ELISA for the Detection of Staphylococcal Enterotoxin A

“…These methods include polymerase chain reaction (PCR), mass spectrometry (MS), biosensor-based techniques, reversed passive latex agglutination (RPLA), immunoblotting, and ELISA [8,16,17,18,19,20]. …”

“…Di Pinto et al compared RPLA and immunoblotting in terms of their capabilities in confirming SEs in culture filtrates of a S. aureus strain [16]. RPLA is a relatively simple method for routine monitoring compared with immunoblotting, which is the standard method.…”

Use of biotin-labeled IgY overcomes protein A interference in immunoassays involving Staphylococcus aureus antigens

“…SEB is a classical enterotoxin and a potent superantigen implicated in food poisoning, and it s also considered a potential bioweapon. Several methods have been reported for detection of SEB in food samples, including PCRs (Abdou et al 2012;Thapa et al 2013), ELISAs (Thompson et al 1986), western blotting, reversed passive latex agglutination (RPLA) testing (Di Pinto et al 2004), quantitative I-PCR, and several biosensor methods. The majority of methods employ antibodies from mammalian sources which react with some of the S. aureus proteins, such as SpA, leading to false positive results.…”

Incidence, growth and enterotoxin production of Staphylococcus aureus in insufficiently dried traditional beef ham “govedja pršuta” under different storage conditions

“…Detection techniques for some of the staphylococcal enterotoxins are commercially available and require from one and a half to 24 h to be completed. Reported detection limits range from 0.5 to 2 ng enterotoxin per gram of food (Di Pinto, Forte, Ciccarese, Conversano, & Tantillo, 2004;Mathieu, Isigidi, & Devriese, 1992;Park, Akhtar, & Rayman, 1994;Vernozy-Rozand, Mazuy-Cruchaudet, Bavai, & Richard, 2004;Wieneke, 1991).…”