Double real time fluorescence PCR detecting method for vancomycin enterococcus

A vancomycin intestinal, real-time fluorescence technology, applied in fluorescence/phosphorescence, measurement device, material analysis by optical means, etc., can solve the problems of low repeatability, cumbersome operation, low sensitivity, etc. Amplification efficiency, the effect of strong PCR amplification ability

Inactive Publication Date: 2007-10-10
YANGTZE RIVER PHARMA GRP BEIJING HAIYAN PHARMA
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Problems solved by technology

At present, the main method of clinical VRE detection is through antibacterial test. This detection method is not only cumbersome, tim

Method used

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Examples

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Embodiment

[0054] Example——Double real-time fluorescent PCR detection of vancomycin-resistant enterococcus and its kit

[0055] 1. Composition of dual real-time fluorescent PCR diagnostic kit for vancomycin-resistant enterococci

[0056] The kit consists of PCR buffer (mix), primer 1, primer 2, two probes, Taq enzyme, UNG enzyme, including two VanA and VanB positive controls and negative controls.

[0057] According to the main factor of clinical enterococcus resistance to vancomycin is caused by two resistance genes, VanA and VanB, we selected the conserved sequences of VanA and VanB genes to design two pairs of primers and two probes, in which the probes were respectively labeled with different wavelengths. fluorescent dyes. The designed Tm values ​​of the two pairs of primers and the two probes are close, with a difference of less than 2°C, so the same parameters can be set for the reaction program.

[0058] Specific primers and probes designed for the VanA gene:

[0059] Specific ...

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Abstract

A double real time fluorescent PCR detecting method of enterococcus resisting vancomycin includes applying fluorescent double-channel detection, utilizing two detection wavelength being not influenced to each other to carry out detection, using wavelength A detection probe FPVabA to label fluorescent dyestuff intensity and using wavelength B detection probe FPVanB to label fluorescent dyestuff intensity, confirming that it is negative if C1 value is zero or greater than 37 and confirming that it is positive if C1 value is less than 36.

Description

technical field [0001] The invention relates to a method for detecting vancomycin-resistant enterococcus by double real-time fluorescent PCR. Background technique [0002] Vancomycin, a glycopeptide antibiotic discovered in the 1950s, was used as the drug of choice for the treatment of multidrug-resistant G+ bacterial infections due to its unique mechanism of action. last line of defense". But with the emergence of Vancomycin Resistant Entorecoccus (VRE), this last line of defense is also in danger of collapsing. Clinically, VRE can not only cause urinary tract infection, abdominal cavity, pelvic infection and surgical wound infection, bacteremia, endocarditis, meningitis, etc., but also transfer its resistance to other pathogenic microorganisms. Therefore, it is not only the VRE itself that poses a serious threat to humans, but also the transfer of their drug-resistant factors will produce some new drug-resistant strains that are difficult to treat. Strengthening VRE det...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N21/00C12Q1/68
Inventor 唐先兵石和鹏
Owner YANGTZE RIVER PHARMA GRP BEIJING HAIYAN PHARMA
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