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Staphylococcus aureus strain PCR (Polymerase Chain Reaction) detection kit

A detection kit and staphylococcus technology, applied in the field of microbial detection, can solve the problems of inaccurate quantification and false positives, and achieve the effects of high sensitivity and specificity, convenient operation, and ensuring reliability.

Active Publication Date: 2013-04-10
泰普生物科学(中国)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Although the conventional PCR method has the advantages of simplicity, rapidity, and sensitivity, it cannot accurately quantify and cause false positives caused by contamination after PCR post-processing.

Method used

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  • Staphylococcus aureus strain PCR (Polymerase Chain Reaction) detection kit
  • Staphylococcus aureus strain PCR (Polymerase Chain Reaction) detection kit
  • Staphylococcus aureus strain PCR (Polymerase Chain Reaction) detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Composition of the kit of the present invention

[0027] Main raw material sources and preparation methods:

[0028] Tris: analytically pure, the product of a qualified supplier, content 99.7%, qualified infrared, pH (5% water) 10.3-10.9, moisture 0.3%, melting point 167-171 ℃, qualified absorption system, qualified maximum impurity content .

[0029] MgCl 2 : Analytical pure, products from qualified suppliers, the content is not less than 99%, the reaction of the aqueous solution is qualified, and the maximum content of impurities is qualified. (MgCl2 is easy to absorb moisture, so store it in a desiccator after using a new bottle).

[0030] EDTA: analytically pure, the product of a qualified supplier, it is a white crystalline powder, soluble in water, the solution is acidic, insoluble in alcohol, the content is not less than 99.5%, the reaction of the aqueous solution is qualified, and the complexing force test is qualified, The highest content of impur...

Embodiment 2

[0070] Example 2 Use of the kit of the present invention

[0071] 1. Reagent preparation:

[0072] 1. Take out 10× concentrated cleaning solution A and 10× concentrated cleaning solution B, dilute with sterile pure water at 1:9 (volume ratio), and put them in a 4°C refrigerator for later use.

[0073] 2. Briefly centrifuge Taq DNA Polymerase and Uracil N-Glycosylase (UNG) and store them in a -20°C refrigerator for later use.

[0074] 3. After determining the number of reaction tubes n (number of samples + negative and positive controls), take out the SA-PCR reaction solution, mix n × 44.3 μl SA-PCR reaction solution / nuc-PCR reaction solution, n × 0.5 μl Taq DNA Polymerase and n×0.2μl Uracil N-Glycosylase (UNG) were added to a centrifuge tube and mixed by shaking. After brief centrifugation, aliquot 45μl into each PCR reaction tube, cover the tube and transfer it to the sample addition area to avoid Store in a 4°C refrigerator for later use.

[0075] 4. Transfer the extracte...

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Abstract

The invention relates to the field of microorganism detection, in particular to a staphylococcus aureus strain PCR (Polymerase Chain Reaction) detection kit. The invention has the technical scheme of providing a staphylococcus aureus strain PCR detection kit which comprises a PCR reaction liquid, and the PCR reaction liquid comprises primers and a fluorescent probe, wherein the primers are divided into an upstream primer and a downstream primer, the nucleotide sequence of the upstream primer is represented by SEQ ID NO.1, and the nucleotide sequence of the downstream primer is represented by SEQ ID NO.2; and the nucleotide sequence of the fluorescent probe is represented by SEQ ID NO.3. The kit has the advantages of high sensitivity, high specificity, stability, timeliness, convenience for operation and the like.

Description

technical field [0001] The invention relates to the field of microorganism detection, in particular to a PCR detection kit for Staphylococcus aureus strains. Background technique [0002] Staphylococcus aureus (SA, referred to as Staphylococcus aureus) is a medically important pathogen with wide distribution, diverse infection types and high drug resistance rate, especially methicillin resistant staphylococcus aureus, MRSA) infection is multi-drug resistant and has a high fatality rate, which has become a difficulty in clinical anti-infective treatment. [0003] Staphylococcus aureus is the most common pathogenic bacteria in human purulent infections, which can cause local purulent infections, pneumonia, pseudomembranous colitis, pericarditis, etc., and even severe systemic infections such as sepsis and sepsis. [0004] The current clinical detection methods of Staphylococcus aureus include: conventional smear microscopy, culture method, immunological assay, instrument auto...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/14G01N21/64
Inventor 何华东姚雪
Owner 泰普生物科学(中国)有限公司