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Staphylococcus aureus methicillin resistant strain PCR (Polymerase Chain Reaction) detection kit

A technology of methicillin-resistant strains and detection kits, applied in the field of microbial detection, can solve problems such as false negatives, achieve high specificity, ensure reliability, and facilitate operation

Active Publication Date: 2012-04-04
福州泰普生物科学有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Latex agglutination reagent is easy to use and fast to detect, but it may be affected by the expression of PBP2α protein, and false negative results may occur

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Composition of the kit of the invention

[0045] Main raw material source and preparation method:

[0046] Tris: analytically pure, the product of a qualified supplier, the content is 99.7%, infrared is qualified, pH (5% water) 10.3-10.9, moisture 0.3%, melting point 167-171℃, the absorption system is qualified, and the highest impurity content is qualified .

[0047] MgCl 2 : Analytical pure, qualified supplier's product, the content is not less than 99%, the aqueous solution reaction is qualified, and the highest impurity content is qualified. (MgCl2 is very easy to absorb moisture, put in a desiccator after opening a new bottle).

[0048] EDTA: analytically pure, the product of a qualified supplier, is white crystalline powder, soluble in water, the solution is acidic, hardly soluble in alcohol, the content is not less than 99.5%, the aqueous solution reaction is qualified, and the complexing power test is qualified. The highest impurity content is qualified.

[0...

Embodiment 2

[0098] Example 2 Use of the kit of the invention

[0099] 1. Reagent preparation:

[0100] 1. Take out 10× concentrated cleaning solution A and 10× concentrated cleaning solution B, dilute them with sterile pure water at a ratio of 1:9 (volume ratio), and put them in a refrigerator at 4°C for later use.

[0101] 2. Centrifuge the Taq DNA Polymerase and Uracil N-Glycosylase (UNG) briefly, and place them in a refrigerator at -20°C for later use.

[0102] 3. After determining the number of reaction tubes n (number of specimens + negative and positive controls) to be carried out, take out the SA-PCR reaction solution / mecA-PCR reaction solution, and add n×44.3μl SA-PCR reaction solution / mecA-PCR reaction solution , N×0.5μl Taq DNA Polymerase and n×0.2μl Uracil N-Glycosylase (UNG) were added to a centrifuge tube and shaken to mix. After instant centrifugation, aliquot 45μl into each PCR reaction tube and cover the tube. Transfer to the sample area and place in a refrigerator at 4°C in the ...

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Abstract

The invention relates to the field of microorganism detection, in particular to a staphylococcus aureus methicillin resistant strain PCR (Polymerase Chain Reaction) detection kit. The invention has the technical scheme of amplifying and detecting a highly conservative specific nucleic acid sequence nuc gene in the staphylococcus aureus gene by using the polymerase chain reaction and molecular beacon and fluorescent probe technologies and simultaneously amplifying and detecting the drug resistant nucleic acid sequence mecA gene to judge whether the strain is a methicillin resistant strain of staphylococcus aureus. The invention also provides PCR amplification primers of the nuc gene and the mecA gene and the sequence of the probe. The kit is applicable to the rapid detection of the staphylococcus aureus methicillin resistant strain in a clinical specimen of a suspect staphylococcus aureus infector and 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 microbial detection, in particular to a PCR detection kit for methicillin-resistant Staphylococcus aureus strains. Background technique [0002] Staphylococcus aureus (Staphylococcus aureus, SA, referred to as Staphylococcus aureus) is a medically important pathogen. It has a wide distribution, various infection types, and a high drug resistance rate. In particular, methicillin resistant staphylococcus Aureus (MRSA) infection is multi-drug resistant and has a high mortality rate, which has become a difficult point in clinical anti-infection treatment. [0003] Staphylococcus aureus is the most common pathogen in human purulent infections. It can cause local purulent infections, pneumonia, pseudomembranous enteritis, pericarditis, etc., and even severe systemic infections such as sepsis and sepsis. [0004] The current clinical application of Staphylococcus aureus detection methods include: conventional smear microscopy, cu...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/14C12R1/445
Inventor 何华东姚雪
Owner 福州泰普生物科学有限公司
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