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Kit for rapidly detecting methicillin-resistant staphylococcus aureus and application thereof

A technology for methicillin-resistant Staphylococcus aureus, applied in the field of rapid detection kits for methicillin-resistant Staphylococcus aureus, to achieve rapid detection, low cost, and high throughput

Pending Publication Date: 2016-03-09
HANGZHOU DIAN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, not all MRSAs carry the mecA gene. Therefore, when identifying MRSA based on the mecA gene, there are often false negative results.

Method used

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  • Kit for rapidly detecting methicillin-resistant staphylococcus aureus and application thereof
  • Kit for rapidly detecting methicillin-resistant staphylococcus aureus and application thereof
  • Kit for rapidly detecting methicillin-resistant staphylococcus aureus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Detection of the nucleic acid sequence of methicillin-resistant Staphylococcus aureus gene detection.

[0033] 2 pairs of specific primers and 2 probes were entrusted to Yingwei Jieji (Shanghai) Trading Co., Ltd. to synthesize:

[0034] F1 (SEQ ID NO: 1): 5'-tataacaacatgaaaaatgattatgg-3';

[0035] R1 (SEQ ID NO: 2): 5'-catatgaaggtgtgcttacaagtgc-3';

[0036] F2 (SEQ ID NO: 3): 5'-gctgatggaaaaatggtaaacga-3';

[0037] R2 (SEQ ID NO: 4): 5'-ctcttttttcgcttgtgcttca-3';

[0038] T1 (SEQ ID NO: 5): Fam-5'-aggtactgctatccaccctcaaacaggtga-3'-tamra;

[0039] T2 (SEQ ID NO: 6): hex-5'-gttcgtcaaggcttggctaaagttgctt-3'-tamra.

Embodiment 2

[0040] Embodiment 2: Reagent preparation.

[0041] Prepare PCR reaction solution components: 10×PCRBuffer, 2mMMgCl 2 , 0.2mMdNTPs, 2U / μL Taq DNA polymerase, SEQ ID NO: 1~620μmol / L, stored at -20℃;

Embodiment 3

[0042]Embodiment 3: detection method.

[0043] Instrument: Roche 480 fluorescent quantitative PCR detector, BECKMAN 22R desktop micro-refrigerated centrifuge, Eppendorf5810R desktop refrigerated centrifuge, Taicang Hualida Laboratory Equipment Company WH-866 vortex oscillator.

[0044] (1) Preparation of bacterial genomic DNA template: refer to published literature, different types of bacterial specimens, prepare bacterial genomic DNA according to the kit instructions, and use it as a PCR reaction template for future use.

[0045] (2) using the genomic DNA described in step (1) as a template, using 2 pairs of specific primers and 2 probes to carry out the amplification detection of the drug-resistant gene mecA of methicillin-resistant Staphylococcus aureus, specifically comprising the following steps;

[0046] (a) Preparation of PCR reaction solution: Take out the components of the kit from the -20°C refrigerator, thaw at room temperature, and put them on an ice box for la...

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PUM

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Abstract

The invention discloses a primer for detecting methicillin-resistant staphylococcus aureus. The primer comprises a primer body for amplifying a methicillin-resistant mecA medicine resistant gene, a primer body for amplifying a staphylococcus aureus nuc gene, a fluorescent probe T1 for detecting the methicillin-resistant mecA medicine resistant gene and a fluorescent probe T2 for detecting the staphylococcus aureus nuc gene. The nucleotide sequence is shown as SEQ ID NO:1-6. The invention further discloses a kit comprising the primer for detecting the methicillin-resistant staphylococcus aureus. The primer and the kit are suitable for rapidly detecting methicillin-resistant staphylococcus aureus strains in a suspicious staphylococcus aureus infected person clinic sample, and have the advantages of being high in sensitivity and specificity, stable, timely, convenient to operate and the like.

Description

technical field [0001] The invention belongs to the technical field of microbial detection, and in particular relates to a rapid detection kit for methicillin-resistant Staphylococcus aureus and an application thereof. Background technique [0002] Staphylococcus aureus (SAU) is one of the most common pathogens causing nosocomial infections. With the widespread use of a large number of broad-spectrum antibacterial drugs in clinical practice, the drug resistance of Staphylococcus aureus in hospital infections is also increasing. In recent years, nosocomial infections The isolation rate of methicillin-resistant Staphylococcus aureus (MRSA) has been at a high level. [0003] The mecA gene is a foreign gene obtained by Staphylococcus aureus through the transposition mechanism, encoding and synthesizing PBP2α. The mecA gene not only mediates the drug resistance of Staphylococcus aureus to β-lactam antibiotics, but also the DNA fragment SCCmec where the mecA gene is located can c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/14C12N15/11C12R1/445
CPCC12Q1/6851C12Q1/689C12Q2531/113C12Q2537/143C12Q2563/107
Inventor 任绪义虞闰六施宏
Owner HANGZHOU DIAN BIOTECH CO LTD
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