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Kit and method for simultaneously detecting Staphylococcus aureus and five enterotoxins thereof

A staphylococcus, a detection method technology, applied in the biological field, can solve the problems of high difficulty in probe design, easy to cause false positive results, high detection cost, etc., to improve detection efficiency and accuracy, shorten detection cycle, and reduce dosage Effect

Active Publication Date: 2017-03-22
SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a detection method for simultaneous detection of Staphylococcus aureus and its 5 kinds of enterotoxins, aiming to solve the problem of high detection cost, high difficulty in probe design and easy detection of 5 kinds of enterotoxins in one reaction tube in the prior art. Problems causing false positive results

Method used

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  • Kit and method for simultaneously detecting Staphylococcus aureus and five enterotoxins thereof
  • Kit and method for simultaneously detecting Staphylococcus aureus and five enterotoxins thereof
  • Kit and method for simultaneously detecting Staphylococcus aureus and five enterotoxins thereof

Examples

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Effect test

Embodiment 1

[0079] A kit for simultaneous detection of Staphylococcus aureus and its five enterotoxins based on fluorescent probe melting curve method. The kit includes:

[0080] 1. Universal primers, such as Q ID NO: 1 and SEQ ID NO: 2 as described in Table 1;

[0081] 2. Fluorescent probe, such as Q ID NO: 3 in Table 1;

[0082] 3. Hybrid connection probes designed according to Staphylococcus aureus and its five enterotoxin-specific genes, among which, Staphylococcus aureus and its five enterotoxins, each Staphylococcus aureus and its five enterotoxin-specific genes The sequences of the ligation probes hybridized with each Staphylococcus aureus and its five enterotoxins are shown in Table 2, respectively.

Embodiment 2

[0084] A kit for simultaneous detection of Staphylococcus aureus and its five enterotoxins based on fluorescent probe melting curve method. The kit includes:

[0085] 1. Universal primers, such as Q ID NO: 1 and SEQ ID NO: 2 as described in Table 1;

[0086] 2. Fluorescent probe, such as Q ID NO: 3 in Table 1;

[0087] 3. According to Staphylococcus aureus and its 5 kinds of enterotoxin-specific gene design hybrid junction probes, wherein, Staphylococcus aureus and its 5 kinds of enterotoxins, their respective specific genes and their respective hybrid junction probe sequences are shown in the table shown in 2.

[0088] 4. The reagent system for the hybridization ligation reaction, as shown in Table 3 above;

[0089] 5. Fluorescent probe melting curve detection reagent system, as shown in Table 4 above.

Embodiment 3

[0091] A kit for simultaneous detection of Staphylococcus aureus and its five enterotoxins based on fluorescent probe melting curve method. The kit includes:

[0092] 1. Universal primers, such as Q ID NO: 1 and SEQ ID NO: 2 as described in Table 1;

[0093] 2. Fluorescent probe, such as Q ID NO: 3 in Table 1;

[0094] 3. According to Staphylococcus aureus and its 5 kinds of enterotoxin-specific gene design hybrid junction probes, wherein, Staphylococcus aureus and its 5 kinds of enterotoxins, their respective specific genes and their respective hybrid junction probe sequences are shown in the table shown in 5.

[0095] 4. The reagent system for the hybridization ligation reaction, as shown in Table 3 above;

[0096] 5. Fluorescent probe melting curve detection reagent system, as shown in Table 4 above.

[0097] table 5

[0098]

[0099]

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Abstract

The present invention provides a kit and a method for simultaneously detecting Staphylococcus aureus and five enterotoxins thereof. The kit comprises a pair of universal primers, a fluorescent probe and a hybridization linking probe designed according to the specific genes of Staphylococcus aureus and five enterotoxins thereof. According to the detection method, the DNA of the suspected bacteria extracted from a sample to be detected is adopted as a template, detection is performed by combining a multiple linking probe amplification technology and a fluorescent probe melting curve technology, and the conditions of the Staphylococcus aureus and the five enterotoxins thereof in the sample to be detected are analyzed and treated through the melting curve.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit and a method for simultaneously detecting Staphylococcus aureus and five kinds of enterotoxins thereof. Background technique [0002] Staphylococcus aureus is one of the most important food-borne pathogens and widely exists in nature. Because Staphylococcus aureus has strong resistance (heat resistance) to various physical and chemical factors, it is easy to contaminate meat and dairy products, so food poisoning incidents caused by Staphylococcus aureus account for a large proportion in countries around the world. Food contaminated by Staphylococcus aureus not only easily leads to spoilage, but also some strains can produce enterotoxin (Staphylococcal enterotoxin, SE). Enterotoxin is the main pathogenic factor causing Staphylococcus aureus food poisoning. It is a toxin protein with low molecular weight (about 26-29kDa) and high thermal stability (boiling at 100°C f...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/14C12N15/11C12R1/445
CPCC12Q1/686C12Q1/689C12Q2600/16C12Q2521/501C12Q2527/107C12Q2537/143C12Q2563/107
Inventor 扈庆华陈清凉姜伊祥石晓路林一曼邱亚群李迎慧江敏陈琼城
Owner SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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