31 results about "STX2" patented technology

The invention provides a quick detecting kit that is used for loop-mediated isothermal augmentation of O157:H7 coliform, designs LAMP primers respectively for O157:H7 O-antigen encoding rfbE, Shiga-like toxin stx2 and H flagellar antigen encoding flic gene, provides a quick detecting kit and a detecting method with strong specificity and high sensitivity that are used for loop-mediated isothermal augmentation of O157:H7 coliform, and has following essential advantages: (1) expensive and precise instruments and equipments are unnecessary, and only a metal pyrolyzer/temperature-constant water bathing device is required; (2) the detecting kit and the detecting method consume less time, and can finish reaction in about 1 hour at constant temperature; (3) a result can be determined by observing color changes of a reaction system with naked eyes under white light, thus having simple and convenient operation; and (4) the method is simple, easy, quick, specific and sensitive, and has wider application prospect in quick detection aspects.

A set of oligonucleotide probe for detecting the enterohemorrhagic E. coli O157:H7 and cholera vibrio O139 is designed on the Shiga-like toxin generating gene stx1 or stx2 and the beta-glucuronidase gene uidA of said enterohemorrhagic E coli O157:H7 and the enterotoxin A subset (ctxA), toxicity coordinate pilus A subset (tcpA) and glucosyltransferase (LPS gt) genes of cholera vibrio O139, and has high sensibility and specificity. It can be used to detect the enterohemorrhagic E. coli O157:H7, cholera vibrio O139 and other pathogenic genes from the specimen.

The invention discloses a kit for rapidly detecting five kinds of diarrheogenic Escherichia coli through multiple qPCR (quantitative Polymerase Chain Reaction). The kit comprises primer and probe groups capable of respectively performing specific amplification on eae genes of EPEC or EHEC, escV genes of EPEC or EHEC, bfpB genes of EPEC, stx1 or stx2 genes of EHEC lt, sth or stp genes of ETEC, aggR, astA or pic genes of EAEC and internal quality control IAC genes. Moreover, the invention further discloses a method and application for rapidly detecting the five kinds of diarrheogenic Escherichiacoli by using the kit. A method for simultaneously detecting the five kinds of diarrheogenic Escherichia coli in foods in the same reaction tube by virtue of fourteen real-time fluorescence quantitative PCR is realized, all characteristic genes in the national standard of the five DEC (diarrheogenic Escherichia coli) can be totally detected, and phenomena such as missing detection and false negative are avoided. The kit has the advantages of being short in detection time, simple and convenient to operate, easily decipherable in results, high in sensitivity, excellent in specificity and the like.

The invention is based on the discovery of the epitope in the Stx2 protein for the 11 E1O antibody. The invention features compositions containing non-full length Stx2 polypeptides that include the 11 E1O monoclonal antibody epitope. The invention also features methods of producing anti-Stx2 antibodies specific for the 11 E1O epitope of the Stx2 protein. Additionally, the invention features methods for treating a subject having, or at risk of developing, a Shiga toxin associated disease (e.g., hemolytic uremia syndrome and diseases associated with E. coli and S. dysenteriae infection) with a polypeptide that includes the 11 E1O epitope or with an anti-Stx2 antibody developed using the methods of the invention. Furthermore, the invention features the detection of Stx2 in a sample using the antibodies developed using the methods of the invention.

The invention discloses a multiplex PCR detection method and kit for Shiga toxin-producing Escherichia coli and application thereof. The method and the kit thereof can simultaneously detect the genes consisting of stx1, stx2 and wzyO157 through a triplex PCR reaction, electrophoresis staining and ultraviolet visualization, subtypes consisting of stx1 and stx1c can be amplified by using a detection primer for the stx1 gene, subtypes consisting of stx2, stx2c and stx2d can be amplified by using a detection primer for the stx2 gene, and an O157 serotype strain can be identified by using a detection primer for the wzyO157 gene. According to the invention, the method and the kit can effectively detect stx and main variants thereof and can be used for distinguishing O157 from other non-O157 Escherichia coli, for directly detecting Shiga toxin-producing Escherichia coli in a food sample and for identifying a clinical strain and a food isolate.

The invention features methods, compositions, and kits for treating a subject having a Shiga toxin associated disease with chimeric anti-Shiga Toxin 1 (cαStx1) and anti-Shiga Toxin 2 (cαStx2) antibodies.

The present invention provides a detection kit for E.coli stx2 gene. The E.coli stx2 gene detection kit includes two pairs of primers designed based on the loop-mediated constant temperature amplification technology with the stx2 gene as the target gene. : inner primer FIP/BIP and outer primer F3/B3. The enterohaemorrhagic Escherichia coli stx2 gene detection kit of the present invention has more comprehensive detection effect and low missed detection rate.

Disclosed are nontoxic mutants of Shiga-like toxin (Stx1 or Stx2), nucleic acids encoding them, compositions containing the mutants and methods of using the mutants in connection with hemolytic euremic syndrome (HUS). Also disclosed are methods of treating HUS using L3 protein fragments, the nontoxic Stx1 or Stx2 mutants, or combinations thereof.

A single kit comprising 3 multiplex PCR assays that can detect in E. coli the presence of the 8 virulence genes: eaeA, EHEC-HlyA, Stx1 (VT1), Stx2 (VT2), Stx2c (VT2c), Stx2d (VT2d), Stx2e (VT2e) and Stx2f (VT2f) is described. In addition, the kit can detect the two critical serotypes (O157 and H7) and identify the species (Escherichia coli) simultaneously using a one step reaction. Following evaluation in our hands, this PCR kit has been used to detect the above 11 components of disease-causing E. coli in a fast, accurate, reliable and specific fashion. These kits can be used on bacterial isolates and has the potential for use directly on foods and environmental samples.

The invention discloses an RPA (recombinase polymerase amplification) primer composition and method for detecting shigella toxin-producing Escherichia coli. The primer composition is used for detecting nine gene subtypes, namely, stx1a, stx1c, stx1d, stx2a, stx2b, stx2c, stx2d, stx2e and stx2g and comprises a primer pair with sequences shown as SEQ ID NO: 1 and SEQ ID NO: 4 and a primer pair withsequences shown as SEQ ID NO: 9 and SEQ ID NO: 12. The method provided by the invention has the advantages of simple operation, high specificity and high sensitivity, is suitable for on-site rapid detection, can complete nucleic acid detection within 20 min, and can provide a rapid screening result for supervision and inspection of listed products and emergency treatment of supervision departments. Meanwhile, the primer and probe combination designed and screened for the first time can effectively detect nine stx gene subtypes, namely stx1a, stx1c, stx1d, stx2a, stx2b, stx2c, stx2d, stx2e andstx2g genes. And in combination with a centrifugal disc type micro-fluidic chip technology, rapid and high-throughput detection on STEC in foods, medicines and livestock breeding products can be realized within an extremely small volume (5 microliters ).

The invention relates to a primer and probe group for detecting enterohemorrhagic escherichia coli, a kit and a PCR (polymerase chain reaction) detection method. Based upon PCR detection on stx1 gene, stx2 gene and eae gene in the enterohemorrhagic escherichia coli, the primer and probe group, which adopts real-time fluorescence PCR detection, is completely consistent with the genotype of an escherichia coli strain in amplification of the strain and is free from specific amplification; therefore, the primer and probe group, which adopts real-time fluorescence PCR detection, has the advantages of being excellent in specificity and real-time fluorescence PCR on on three genes in the enterohemorrhagic escherichia coli, namely the stx1 gene, the stx2 gene and the eae gene; and meanwhile, the corresponding PCR detection method has the characteristics of being strong in specificity, high in sensitivity, rapid and reliable in detection and simple and convenient to operate.

The invention discloses a CPA detection primer for escherichia coli shiga toxin type II, a kit and a method thereof. The CPA detection primer is designed for the specific target sequence stx2 of the escherichia coli shiga toxin type II, a primer sequence is shown as SEQ ID NO.1-5, and the primer ensures the reliability of the detection result. The invention further provides a CPA detection kit forthe escherichia coli shiga toxin II type. The CPA detection kit has the advantages of high sensitivity, good applicability, good specificity, simplicity, convenience and rapidness in operation, accurate and reliable result, low detection cost and the like. The CPA detection primer or the CPA detection kit is used for carrying out a cross primer isothermal amplification reaction on a to-be-detected sample, a result can be directly judged and read by naked eyes through fluorescent dye, whether the to-be-detected sample contains the escherichia coli shiga toxin type II or not can be rapidly andaccurately detected, and the kit is particularly suitable for small and medium-sized units and field detection.

Human and humanized monoclonal antibodies which binds specifically to subunit A of Shiga like toxin II have been developed which are effective to prevent or ameliorate one or more symptoms of HUS in a human. Effective dosages for treatment or prevention range from approximately 0.1 to 5.0 mg of antibody/kg of patient weight. The examples demonstrate the preferred dosage ranges based on the pig model, and what is being tested in phase I clinical trials. Antibodies are preferably transfused over a period of two hours, although this will depend on the patient and the disease state at the time of treatment. Preferred dosages for treatment of humans are between 0.1 mg/kg-5.0 mg/kg of 5C120, or an equivalent dosage of another antibody to subunit A of STX2. In the most preferred embodiments, dosages of 0.1 mg/kg, 0.5 mg/kg, or 5.0 mg/kg of 5C12 (low dose, anticipated therapeutic dose based on animal data and high dose) are administered.