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Nucleic acid reagent, kit, system and method for detecting Gram-negative bacteria and/or drug resistance of Gram-negative bacteria

A Gram-negative bacteria, Gram-negative technology, applied in the field of nucleic acid reagents for detecting Gram-negative bacteria and/or Gram-negative bacteria drug resistance

Active Publication Date: 2019-03-29
北京卓诚惠生生物科技股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although these detection methods can systematically and comprehensively identify drug resistance genes, they all have their own technical drawbacks.

Method used

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  • Nucleic acid reagent, kit, system and method for detecting Gram-negative bacteria and/or drug resistance of Gram-negative bacteria
  • Nucleic acid reagent, kit, system and method for detecting Gram-negative bacteria and/or drug resistance of Gram-negative bacteria
  • Nucleic acid reagent, kit, system and method for detecting Gram-negative bacteria and/or drug resistance of Gram-negative bacteria

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Embodiment

[0066] 1. Primer and probe synthesis

[0067] Sequence synthesis was performed according to the primer sequences shown in Table 1 and the probe sequences shown in Table 2. In the probe, FAM is 6-carboxyfluorescein, JOE is 2,7-dimethyl-4,5-dichloro-6-carboxyfluorescein, CY5 is 5H-indocyanine, and ROX is 6-carboxy-X-rhodan It is indicated that VIC is a dye purchased from ABI Company. The brackets in the probe sequences in Table 2 indicate that the bases on the left side of the brackets have fluorescent labels, and the content in the brackets indicates the choice of fluorescent labels.

[0068] Table 1

[0069]

[0070]

[0071] Table 2

[0072]

[0073]

[0074] 2. Sample processing

[0075] After the patient's anal swab sample is collected by conventional methods, it is processed with a commercial extraction kit.

[0076] Use the sampler matched with ParaDNA to collect the processed anal swab samples, and place them directly in the ParaDNA reactor for amplifica...

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Abstract

The invention relates to a nucleic acid reagent, a kit, a system and a method for detecting Gram-negative bacteria and / or drug resistance of Gram-negative bacteria. The nucleic acid reagent comprisesprimers shown as the SEQ ID NO. 1-60 and probes shown as the SEQ ID NO. 63-92 which are independently stored or stored as arbitrary mixtures. By using the primers and the probes, the nucleic acid reagent, the kit, the system and the method for detecting 6 pathogens, including Klebsiella pneumoniae, Escherichia coli, pseudomonas aeruginosa, enterobacter cloacae, serratia marcescens and acinetobacter baumannii, as well as 24 common drug resistance genes carried by the 6 pathogens are established by the invention; moreover, fast, comprehensive, sensitive, specific and automatic detection result determination can be realized, so that, sensitivity, specificity and simplicity of simultaneous detection of multiple Gram-negative bacteria and drug resistance thereof are significantly improved.

Description

technical field [0001] The present disclosure relates to the field of biotechnology, in particular, to a nucleic acid reagent, kit, system and method for detecting Gram-negative bacteria and / or Gram-negative bacteria drug resistance. Background technique [0002] Gram-negative bacteria accounted for 60% of the spectrum of pathogens causing infections, mainly Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Bacillus cloacae, Serratia marcescens, Acinetobacter baumannii bacilli. With the extensive use of antibiotics, Gram-negative bacteria have gradually developed resistance. Drug resistance, also known as drug resistance, generally refers to the reduction or even disappearance of the sensitivity of bacteria to drugs after repeated contact with drugs, resulting in reduced or ineffective efficacy of drugs against drug-resistant bacteria. After the emergence of drug-resistant bacteria, they quickly spread among different regions, between the same species and di...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/10C12Q1/04C12N15/11C12M1/34C12M1/00C12R1/22C12R1/19C12R1/385C12R1/43C12R1/01
CPCC12Q1/689C12Q2600/16C12Q2600/166
Inventor 王雷杨海英林笑冬王晓艳张志强
Owner 北京卓诚惠生生物科技股份有限公司
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