Authenticating gram positive bacteria species and method for testing drug resistant gene and dedicating kit

A Gram-positive, detection kit technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of not targeting special populations, single molecular biology detection indicators, and poor practicability.

Inactive Publication Date: 2008-10-22
CAPITALBIO CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, conventional molecular biology detection indicators are single, so the practicability is poor
[0013] 2. The current main problem of drug resistance of Gram-positive bacteria
[0034] 1. It can only be used for the identification of bacterial species (such as Li Junwen et al., an oligonucleotide probe for simultaneous detection of multiple bacteria, application number 01120290.4; Li Junwen et al., a DNA microprobe for detecting common pathogenic bacteria in water. Array, application number 01120291.2; Ma Wei et al., biochip for diagnosis of common clinical pathogenic bacteria based on 16S rDNA gene species-specific sequence, application number 02136567.9) or bacterial drug resistance detection (Liu Yuan et al., using gene chip technology to detect drug resistance genes, Application No. 01120441.9)
[0035] 2. Wide range of requirements, no special population, poor practicability
[0036] 3. These patents do not provide a set of effective and specific detection schemes, so their feasibility is poor

Method used

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  • Authenticating gram positive bacteria species and method for testing drug resistant gene and dedicating kit
  • Authenticating gram positive bacteria species and method for testing drug resistant gene and dedicating kit
  • Authenticating gram positive bacteria species and method for testing drug resistant gene and dedicating kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0110] Embodiment 1, utilize Gram-positive bacterial species identification and drug-resistant gene detection kit of the present invention to carry out staphylococcus, enterococcus, streptococcus species identification and drug-resistant gene detection

[0111] 1. Detection targets, primers and probes

[0112] The detection targets of this kit include: Staphylococcus aureus, coagulase-negative staphylococcus of the genus Staphylococcus, Enterococcus faecalis and Enterococcus faecium of the genus Enterococcus, Streptococcus pneumoniae of the genus Streptococcus, and their drug resistance genes mecA, aac(6')-Ie-aph(2")-Ia, vanA, vanB, pbp-la / pbp-2b / pbp-2x, ermA, ermB, ermC, mefA and msrA.

[0113] Select primers and probes corresponding to the detection target, as shown in Table 3 and Table 4 (detailed information is shown in Table 1 and Table 2).

[0114] Table 3 Gram-positive bacteria identification and drug resistance detection gene chip kit primers (31 in total)

[0115] ...

Embodiment 2

[0153] Embodiment 2, using the gram-positive bacterial species identification and drug-resistant gene detection kit of the present invention to carry out enterococcus identification and drug-resistant gene detection

[0154] 1. Detection targets, primers and probes

[0155] The detection targets of this kit include: Enterococcus faecalis, Enterococcus faecium, Enterococcus faecium and Enterococcus durascens and Enterococcus hirae, Enterococcus gallinarum and Enterococcus avium, Enterococcus japonica and Enterococcus saccharolyticus of the Enterococcus genus , and its resistance genes aac(6')-Ie-aph(2")-Ia, aadE, vanA and vanB.

[0156] Select primers and probes corresponding to the detection target, as shown in Table 8 and Table 9 (detailed information is shown in Table 1 and Table 2).

[0157] Table 8 Enterococcus identification and drug resistance detection gene chip kit used primers (15 in total)

[0158]

[0159]

[0160] Table 9 Enterococcus identification and dru...

Embodiment 3

[0177] Embodiment 3, use Gram-positive bacterial species identification and drug-resistant gene detection kit of the present invention to carry out staphylococcus species identification and drug-resistant gene detection

[0178] 1. Detection targets, primers and probes

[0179] The detection targets of this kit include: Staphylococcus aureus of the genus Staphylococcus, coagulase-negative staphylococci, and their drug resistance genes mecA, blaZ, aac(6')-Ie-aph(2")-Ia, ermA / ermC and msrA.

[0180] Select primers and probes corresponding to the detection target, as shown in Table 11 and Table 12 (detailed information is shown in Table 1 and Table 2).

[0181] Primers and probes were synthesized by Shanghai Boya Biotechnology Company. The TAMRA fluorescent label at the 5' end of some primers and the amino modification at the 5' end of the probe were also completed by Shanghai Boya Biotechnology Company.

[0182] 2. Identification of staphylococcus and preparation of gene chi...

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PUM

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Abstract

This invention discloses a method of authenticating species of gram positive bacterium and examining drug resistant gene, special reagent kit. This kit contains species special gene of amplified gram positive bacterium and antithetical couplet of primer gene and gene probe of gram positive bacterium and anti-drug. This method of authentication of species and examination of drug-resistant gene consists of these steps: 1. amplify the species gene and anti-drug gene by the primer in the box. 2. Hybridize said two gene probes. This invention of kit has high integrated degree, high sensitivity, and broad application, and high specific activity, stable and reliable detecting signal.

Description

technical field [0001] The invention relates to a Gram-positive bacterial species identification method and a drug resistance gene detection method and a special kit thereof. Background technique [0002] 1. Significance of bacterial identification and drug resistance detection [0003] Bacteria are the pathogens that cause most infectious diseases. The pathogenic bacteria that cause clinical bacterial infections include aerobic bacteria and anaerobic bacteria, which are divided into two categories: Gram-positive bacteria and Gram-negative bacteria. Bacteria and bacilli. Aerobic Gram-positive cocci and Gram-negative bacilli are the most common pathogenic bacteria in clinical practice. Gram-positive bacteria and Gram-negative bacteria have significant differences in their mechanism of action and drug resistance due to the difference in their cell wall and other structures. Clinically, there are great differences in the clinical use of these two types of bacteria. [0004] ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 张治位祝令香王璨杨华卫张琼程京
Owner CAPITALBIO CORP
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