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Integron In0

A technology of integrons and strains, applied in the field of integrons In0, to reduce selection pressure and prevent explosive epidemics

Inactive Publication Date: 2015-04-22
王冬国
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Antibacterial drugs have played an important role in controlling Escherichia coli infection, but with the widespread use of antibacterial drugs, drug-resistant strains of Escherichia coli, especially multi-drug-resistant strains, continue to appear. important factor for industry development

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Identification of integron In0

[0028] 1. Isolation and identification of strain EC6335

[0029] 1.1 Materials

[0030] Bacterial susceptibility card: AST-GN13 from bio Merieux, France. AST-GN13 drug-sensitive types include: amikacin, ampicillin, ampicillin / sulbactam, aztreonam, cefazolin, cefepime, cefotetan, ceftazidime, ceftriaxone, ciproxa Star, ertapenem, gentamicin, imipenem, levofloxacin, nitrofurantoin, piperacillin / tazobactam, tobramycin, SMZ.

[0031] Supplementary drug-sensitivity disc (drug-sensitivity plate agar diffusion test): the disc comes from Oxoid Company in the United Kingdom, with cefoperazone / sulbactam (75 μg / 30 μg).

[0032] 1.2 Method

[0033] Apparatus identification: transfer positive bacterial strains from the pus of patients in the intensive care unit of Taizhou Municipal Hospital to the blood plate for isolation and culture (at 35°C containing 5% CO 2 Cultivate in the incubator for 16-18h), and then carry out bacterial ident...

Embodiment 2

[0049] Example 2 Plasmid transduction experiments to study the function of integron In0

[0050] 1. Method

[0051](1) The donor bacterium is EC6335 strain, and the recipient bacterium is E.coli J53AzR (resistance to sodium azide). The donor bacteria and recipient bacteria were inoculated on LB plates, respectively, and cultured overnight at 35°C. Pick a single colony and inoculate them in 4 mL of LB broth, and culture at 37°C and 220 r / min until the logarithmic growth phase. Take 0.5ml of donor and recipient bacteria in 4ml of LB broth, and culture overnight at 37°C. Zygotes were screened on trypan soy agar (TSA) plates containing sodium azide (300 mg / L) and imipenem (2 mg / L). Incubate at 35°C for 18-24h. The plasmid extracted from the drug-resistant strain (the steps are the same as in Example 1) was used as a template for PCR amplification, and the primers in Table 1 were used to detect the presence of the integron gene sequence.

[0052] (2) Select positive bacterial ...

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Abstract

The invention discloses a new integron In0 with a sequence shown by SEQ ID NO.1. The integron is discovered in a genome of drug-resistant Escherichia coli EC6335. Therefore, on the one hand, the discovery of the integron disclosed by the invention has important theoretical and practical significances for discussing a molecular mechanism of bacterial drug resistance transfer from a gene level, and on the other hand, the discovery of the integron provides a certain guiding function for clinical medication, and is beneficial for clinically reducing the use of certain antibacterial drugs, reducing the selection pressure of horizontal transfer of drug-resistant gene cassettes, and preventing the explosive prevailing of drug-resistant bacteria.

Description

technical field [0001] The invention belongs to the field of molecular biology technology and drug-resistant bacteria monitoring, and relates to a newly discovered integron In0 closely related to drug resistance of Escherichia coli. Background technique [0002] Escherichia coli is one of the most common pathogenic bacteria in medicine and veterinary clinics, which has caused serious damage to human health and the development of animal husbandry. Antibacterial drugs have played an important role in controlling Escherichia coli infection, but with the widespread use of antibacterial drugs, drug-resistant strains of Escherichia coli, especially multi-drug-resistant strains, continue to appear. important factor for industry development. [0003] Bacterial drug resistance mechanism - integron (integron) system has been widely noticed by researchers, and great progress has been made. Through the integron system, the bacteria capture the foreign drug resistance gene through the ...

Claims

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Application Information

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IPC IPC(8): C12N15/70C12N1/21C12N15/10C12R1/19
Inventor 王冬国梁海燕牟永华陈佳玉
Owner 王冬国
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