Shiga toxins ELISA Kit

Name: Shiga toxins ELISA Kit
SKU: DEIASL162
Rating: 5 (1 reviews)

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

COA

SDS

Datasheet

Summary

Sample

Fecal

Species Reactivity

Human

Detection Method

sELISA

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.

Performance Characteristics

Direct Fecal Testing
The performance of the SHIGA TOXIN test was compared to the Vero Cell Cytotoxin Assay (with neutralization), considered the clinical reference standard (gold standard), and included 899 fresh and 14 frozen specimens. Age information was available for 902 patients. Of the 902 patients, 8.8% were ≤ 18 years. The following table shows a summary of the clinical performance of the SHIGA TOXIN test. The results show that the test exhibited a sensitivity of 100% a speciffcity of 99.9% and an overall correlation of 99.9% with the cytotoxin assay.
Direct Fecal Testing Results

Broth Cultures
The performance of the SHIGA TOXIN test using overnight broth cultures (GN or MacConkey broth) from fecal specimens was compared to the Vero Cell Cytotoxin Assay (with neutralization), considered the clinical reference standard (gold standard). The following table shows a summary of the clinical performance of the SHIGA TOXIN test. The results show that the test exhibited a sensitivity of 97.1%, a speciffcity of 99.7% and an overall correlation of 99.5% with the cytotoxin assay.
Broth Culture Testing Results

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. STEC cause foodborne and waterborne diarrheal disease worldwide which, if left undiagnosed, can progress to hemorrhagic colitis and/or hemolytic uremic syndrome (HUS). Since certain treatments and medications can increase the risk of HUS, prompt detection is necessary to prevent outbreaks and secondary transmission. STEC strain O157:H7 has historically been the focus of attention in the United States since first isolated from undercooked hamburgers, causing an estimated 73,000 illnesses annually. However, STEC infections caused by non-O157 strains have become more prevalent in recent years, both in the United States as well as abroad. O157:H7 infections are routinely diagnosed by culture of fecal samples on selective media, but this methodology allows nonO157 STEC strains to go undetected. STEC produce either one or both Shiga toxins (Stx1 and/ or Stx2), both potent cytotoxins. Isolates producing only Stx2 have been attributed to higher incidence rates of HUS. Shiga toxins can be detected by tissue culture assay, 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.

Citations

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Parallel detection of okadaic acid and saxitoxin in mussels with an electrochemical aptamer-based assay

Szymczyk-Drozd A, Selvolini G, Carbone M, Marrazza G

Talanta2026 Mar 1PubMed ID:41275719Read Article

Applications: ELISA
Reactive species: STX
"Abstract: Phycotoxins are highly toxic compounds bioaccumulating in marine organisms, thus seriously affecting human health. Two electrochemical aptasensors for parallel detection of okadaic acid (OA) and saxitoxin (STX), exploiting a competitive strategy, are then proposed in this study. Screen-printed electrochemical cells were modified first by the electrodeposition of gold nanoparticles and then by the electropolymerization of poly(aniline-co-anthranilic acid) copolymer or poly(l-lysine) film. A conjugate between bovine serum albumin and each phycotoxin was immobilized covalently on the polymeric film and competed with the free phycotoxin in solution for the binding with a specific biotinylated aptamer. The competition was traced with an enzymatic conjugate between streptavidin and alkaline phosphatase: upon addition of 1-naphthyl phosphate, the enzymatic conversion of the substrate occurred. Differential pulse voltammetry (DPV) measurements were carried out to detect the enzymatic product 1-naphthol. A signal-off response was retrieved with detection limits of 66.4 pg/mL for OA and 7.9 pg/mL for STX, respectively. The aptasensors were also tested in the analysis of real mussel samples and their performance was compared with those of commercial enzyme-linked immunosorbent assays. The developed sensing strategy offers an efficient approach for monitoring OA and STX in seafood, thus representing a significant step towards ensuring consumer safety."
Article snippet: Okadaic acid-bovine serum albumin conjugate (OA-BSA) solution, saxitoxin-bovine serum albumin conjugate (STX-BSA) solution and the enzyme-linked immunosorbent assay (ELISA) kits for OA and STX were purchased from Creative Diagnostics (Shirley, NY, USA).

Regarding OA detection, higher recovery values were retrieved for the aptasensors compared to ELISAs, while this trend is inverted in the case of STX. However, in almost all cases, an acceptable bias was found for spiked mussel samples, assessing the applicability of the aptasensors in real samples. (Szymczyk-Drozd A, <i>et al</i>, 2026)

Figure 1. Selectivity of the developed sensors for both a) okadaic acid (OA) (5 ng/mL each) and b) saxitoxin (STX) (3 ng/mL each) in mussel extract.

Precise virulence inactivation using a CRISPR-associated transposase for combating Enterobacteriaceae gut pathogens

Ronda C, Perdue T, Schwanz L, Rivera Gelsinger D, Brockmann L, Kaufman A, Huang Y, Sternberg SH, Wang HH

Nat Biomed Eng2025 DecPubMed ID:40681864Read Article

Applications: ELISA
Reactive species: S. dysenteriae
"Abstract: Targeted gene manipulation in a complex microbial community is an enabling technology for precise microbiome editing. Here we introduce BACTRINS, an in situ microbiome engineering platform designed for efficient and precise genomic insertion of a desired payload and simultaneous knockout of target genes. This system leverages conjugation-mediated delivery of CRISPR-associated transposases to achieve RNA-guided genomic integration, allowing precise insertion of a therapeutic payload while neutralizing pathogen virulence without causing cell death. When applied against an Enterobacteriaceae Shiga toxin-producing pathogen in the gut, this system delivers a CRISPR-associated transposase by bacterial conjugation for site-specific inactivation of the Shiga toxin gene and integration of a nanobody therapeutic payload to disrupt pathogen attachment. A single dose of this therapy results in high-efficiency Shiga gene inactivation and improved survival in a murine infection model of Shiga-producing pathogen. This work establishes a new type of live bacterial therapeutic capable of reducing gut infections by transforming toxigenic pathogens into commensal protectors."
Article snippet: Shiga toxin levels were determined using an ELISA kit for simultaneous detection of Stx1 and Stx2 in a single test (Creative Diagnostics, DEIASL162).

ELISA quantification (absorbance 450–620 nm) of Shiga toxin levels in pathogen culture supernatants, comparing wild-type (WT) pathogens and their derivatives carrying BTS insertions (Stx1A::Tn or Stx2B::Tn). (Ronda C, <i>et al</i>, 2025)

Figure 1. In vitro validation of the BACTRINS system.

Background

Shiga toxins (Stx) are a family of potent bacterial toxins produced by certain strains of Shigella dysenteriae and enterohemorrhagic Escherichia coli (EHEC), including the notorious E. coli O157:H7. These toxins are named after Kiyoshi Shiga, a Japanese bacteriologist who isolated and studied a gram-negative bacteria called Bacillus dysenteriae. He identified this bacteria during a dysentery epidemic in the nineteenth century by isolating it from the stools of affected patients.
Stx are composed of two subunits, A and B. The B subunit is responsible for recognizing and binding to specific receptors on the surface of target cells. The A subunit possesses enzymatic activity and exerts cytotoxic effects on cells. Stx are notorious for their ability to inhibit protein synthesis in host cells, leading to cell death and tissue damage. One of the key receptors recognized by Stx is globotriaosylceramide (Gb3), which is expressed on the surface of certain cells, particularly in the kidneys and the endothelial lining of blood vessels. The binding of Stx to Gb3-expressing cells triggers a cascade of events that can lead to severe clinical manifestations, including hemolytic uremic syndrome (HUS), a life-threatening condition characterized by hemolytic anemia, thrombocytopenia, and acute kidney injury.
Shiga toxin (Stx) detection in fecal samples holds significant importance in clinical and public health settings. By identifying Stx in feces during outbreaks, the source of contamination can be traced, aiding in targeted interventions and preventing further cases. Monitoring Stx clearance in fecal samples helps assess treatment response and guide patient management decisions. Fecal Stx detection contributes to surveillance efforts, enabling the monitoring of prevalence, identifying high-risk groups, and implementing preventive measures. Shiga toxins ELISA kits provide a sensitive and specific method for detecting and quantifying Shiga toxins in clinical samples. They are valuable tools for diagnosing Shiga toxin-producing bacterial infections, monitoring disease progression, and assessing the effectiveness of treatments.

Alternative Names

Shiga toxins
Shiga toxin (Stx)
Shiga toxin ELISA Kit
Shiga toxin (Stx) ELISA Kit

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QA & Reviews

Customer Reviews

References

Shiga toxins—from cell biology to biomedical applications

Nature Reviews Microbiology

Authors: Johannes L, Römer W.

Abstract

Shiga toxin-producing Escherichia coli is an emergent pathogen that can induce haemolytic uraemic syndrome. The toxin has received considerable attention not only from microbiologists but also in the field of cell biology, where it has become a powerful tool to study intracellular trafficking. In this Review, we summarize the Shiga toxin family members and their structures, receptors, trafficking pathways and cellular targets. We discuss how Shiga toxin affects cells not only by inhibiting protein biosynthesis but also through the induction of signalling cascades that lead to apoptosis. Finally, we discuss how Shiga toxins might be exploited in cancer therapy and immunotherapy.

Pathogenesis of Shiga-Toxin Producing Escherichia coli

Ricin and Shiga Toxins: Pathogenesis, Immunity, Vaccines and Therapeutics

Authors: Melton-Celsa A, Mohawk K, Teel L, et al.

Abstract

Shiga toxin (Stx)-producing Escherichia coli (STEC) are food-borne pathogens that cause hemorrhagic colitis and a serious sequela, the hemolytic uremic syndrome (HUS). The largest outbreaks of STEC are due to a single E. coli serotype, O157:H7, although non-O157 serotypes also cause the same diseases. Two immunologically distinct Stxs are found in E. coli, Stx1 and Stx2. The Stxs are AB5 toxins that halt protein synthesis in the host cell, a process that may lead to an apoptotic cell death. Stx-mediated damage to renal glomerular endothelial cells is hypothesized as the precipitating event for HUS. A subset of STEC referred to as the enterohemorrhagic E. coli has the capacity to intimately attach to and efface intestinal epithelial cells, a pathology called the A/E lesion. The A/E lesion is mediated by the adhesin intimin, its bacterially encoded receptor, Tir, and effectors secreted through a type III secretion system. The proteins needed for the A/E lesion are encoded within a large pathogenicity island called the locus of enterocyte effacement or LEE. There are several animal models for STEC infection, but no one model fully represents the spectrum of STEC illness. Currently there is no cure for STEC infection, and therapies are based mainly on alleviating symptoms. However, chimeric or humanized monoclonal antibodies have been developed that neutralize the Stxs, and those therapies may be able to prevent the development of HUS in an STEC-infected patient.

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.

Abstract

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.

Abstract

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.