Shiga toxins ELISA Kit (DEIASL162)

Regulatory status: For research use only, not for use in diagnostic procedures.

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Size
96T
Sample
Fecal
Species Reactivity
Human
Intended Use
The Shiga toxins ELISA Kit is an enzyme immunoassay for the simultaneous qualitative detection of Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2) in a single test. It is intended for use with human fecal samples from patients with gastrointestinal symptoms to aid in the diagnosis of disease caused by Shiga Toxin producing Escherichia coli (STEC). It may be used directly with human fecal specimens, or broth or plate cultures derived from fecal specimens.
Storage
The kit should be stored between 2°C and 8°C. The kit containing the reagents with designated shelf life should be stored between 2°C and 8°C and should be returned to the refrigerator as soon as possible after use.
Precision
Intra-Assay
For the determination of intra-assay performance, 6 positive fecal specimens and 6 negative fecal specimens were analyzed. Each specimen was assayed in replicates of eight. All positives remained positive and all negatives remained negative.
Inter-Assay
The inter-assay precision of the Shiga toxins ELISA Kit was determined using 12 fecal specimens (six negative, two positive for Stx1, two positive for Stx2, and two positive for both Stx1 and Stx2). The samples were tested, twice a day over a 5-day period using 2 different kit lots. A positive and negative control was run on each day. All positives remained positive and all negatives remained negative.
Sensitivity
The cutoff for the test for direct fecal specimens was established at concentrations of 0.28 ng/mL Stx1 and 0.23 ng/mL Stx2, and for broth cultures at concentrations of 0.18 ng/mL Stx1 and 0.30 ng/mL Stx2.
General Description
Shiga toxin producing Escherichia coli (STEC) were first described by O' Brien, et al. after discovering that E.coli culture supernatant, which was cytotoxic to HeLa and Vero cells, could be neutralized by rabbit anti-Shiga toxin antibodies (1). STEC cause foodborne and waterborne diarrheal disease worldwide which, if left undiagnosed, can progress to hemorrhagic colitis and/or hemolytic uremic syndrome (HUS) (2, 3). Since certain treatments and medications can increase the risk of HUS (4), prompt detection is necessary to prevent outbreaks and secondary transmission (5-9). STEC strain O157:H7 has historically been the focus of attention in the United States since first isolated from undercooked hamburgers (3, 10), causing an estimated 73,000 illnesses annually (11). However, STEC infections caused by non-O157 strains have become more prevalent in recent years, both in the United States as well as abroad (12-16, 29). O157:H7 infections are routinely diagnosed by culture of fecal samples on selective media (17, 18), but this methodology allows non-O157 STEC strains to go undetected. STEC produce either one or both Shiga toxins (Stx1 and/ or Stx2), both potent cytotoxins (19, 20). Isolates producing only Stx2 have been attributed to higher incidence rates of HUS (18, 21-23). Shiga toxins can be detected by tissue culture assay (24), but this method is both time consuming and labor intensive. By detecting the toxins, the SHIGA TOXIN CHEK test can detect STEC present in fecal samples or culture, regardless of the serotype or other virulence factors (25).

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References


Molecular characteristics and genotypic diversity of enterohaemorrhagic Escherichia coli O157:H7 isolates in Gauteng region, South Africa

SCIENCE OF THE TOTAL ENVIRONMENT

Authors: Bolukaoto, John Y.; Kock, Marleen M.; Strydom, Kathy-Anne; Mbelle, NontombiM.; Ehlers, Marthie M.

Enterohaemorrhagic Escherichia toff (EHEC) O157:H7 is one of the major foodbome and waterborne pathogens causing severe diseases and outbreaks worldwide. There is scarcity of EHEC O157:H7 data in South Africa. This study was carried out to determine the molecular characteristics and genotypic diversity of EHEC O157:H7 isolates in the Gauteng region, South Africa. Samples were cultured on selective chromogenic media. Antibiotic susceptibility profile of isolates was determined using the VITEK (R)-2 automated system. Isolates were characterised using multiplex PCR assays and the genetic diversity was determined using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MIST). A total of 520 samples of which 270 environmental water samples and 250 stool specimens were collected and analysed. Overall, EHEC O157:H7 was recovered from 2.3% (12/520) of samples collected. Environmental water samples and clinical stool specimens showed a prevalence of 4.07% (11/270) and 0.4% (1/250) respectively. Antibiotic susceptibility profile varied from isolates with full susceptibility to isolates with resistance to multiple antibiotics. Most resistance was detected to the penicillins, specifically ampicillin (7/12). amoxicillin (3/12) and piperacillin/Tazobactam (3/12) followed by one of the folate inhibitors, trimethoprim (3/12) and the carbapenems, imipenem and meropenem (2/12) each. Three isolates harboured a combination of Shiga-toxins (Six)-2, intimin (eae) and enterohaemolysin (hlyA) genes, while two isolates harboured the Stx-1, Slx-2 and hlyA genes. The PFGE performed showed that EHEC O157:H7 isolates were genetically diverse, with two minor pulsotypes and eight singletons. The MIST analysis identified sequence types (STs) (ST10, ST11 and ST1204) that have been previously reported associated with outbreaks. The STs identified in this study pose a potential public health risk to consumers of untreated environmental water and dosed human contacts. There is necessity to enhance surveillance in reducing the propagation of this bacterium which is a public health problem. (C) 2019 Elsevier B.V. All rights reserved.

Interactions of Shiga toxin-producing Escherichia coli with leafy green vegetables

BRAZILIAN JOURNAL OF MICROBIOLOGY

Authors: Abe, Cecilia M.; Matheus-Guimaraes, Cecilia; Garcia, Bruna G.; Cabilio Guth, Beatriz E.

Shiga toxin-producing Escherichia coli (STEC) are important foodborne pathogens responsible for a wide spectrum of diseases including diarrhea, bloody diarrhea, and hemolytic uremic syndrome (HUS). A considerable number of outbreaks and sporadic cases of HUS have been associated with ingestion of fresh ready-to-eat products. Maintenance and persistence of STEC in the environment and foods can be related to its ability to form biofilm. A non-O157 STEC strain isolated from bovine feces was distinguished by its great ability to form biofilm in abiotic surfaces. In the present study, we aimed to investigate the ability of this strain to adhere to rocket leaves (Eruca sativa). Adherence assays were carried out for 3 h at 28 degrees C and analyzed by scanning electron microscopy. The non-O157 STEC strain adhered to leaf surface and inside the stomata forming several bacterial aggregates. The number of adherent bacteria per square millimeter of leaf was eightfold higher compared with an O157 STEC strain. Deletion of the STEC autotransporter protein contributing to biofilm (Sab) reduced the adherence ability of the non-O157 strain in almost 50%, and deletion of antigen 43 (Ag43) almost abolished this interaction. Very few bacteria were seen on the leaf surface, and these differences were statistically significant, suggesting the role of both proteins and especially Ag43 in the interaction of the non-O157 STEC strain with leaves. The risk posed by non-O157 STEC adherence to leaves on fresh produce contamination should not be neglected, and measures that effectively control adherence should be included in strategies to control non-O157 STEC.

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