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Application of specific genes of staphylococcus epidermidis

A technology of staphylococcus epidermidis and specific genes, applied in the biological field, can solve the problems of low sensitivity, narrow application range, and inability to achieve rapid goals, etc.

Active Publication Date: 2015-09-09
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the detection methods of staphylococcus epidermidis in clinical application include conventional smear microscopy, culture method, immunological determination, instrument automatic analysis and identification system, etc., but these methods have certain defects, such as conventional smear microscopy is the most basic The current bacteriological examination method has low sensitivity and poor specificity; the culture method is currently the main way and means to find the source of infection clinically, but it is very time-consuming and cannot achieve rapid goals; Salt content has high requirements. If the salt content is relatively high, the synthesis of protein will be inhibited and false negatives will appear. Expensive

Method used

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  • Application of specific genes of staphylococcus epidermidis
  • Application of specific genes of staphylococcus epidermidis
  • Application of specific genes of staphylococcus epidermidis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1: Specific target gene screening and primer design

[0016] Local BLAST analysis

[0017] From the complete genome sequence of Staphylococcus epidermidis downloaded from NCBI (http: / / www.ncbi.nlm.nih.gov / genome / ), local BLAST search was performed on the local nucleic acid database to obtain the sequence fragments and databases of Staphylococcus epidermidis Compare the results.

[0018] BLAST reconfirmation

[0019] Using the online BLAST function, 2 strategies were used to confirm their species specificity and S. epidermidis identification availability. One strategy is to exclude Staphylococcus epidermidis during BLAST, and the results show that those without any similar sequences are specific genes of Staphylococcus epidermidis; the second strategy is to search within Staphylococcus epidermidis when BLAST returns a large number of similar sequences. Staphylococcal strain consensus sequences. Combining the two strategies, we finally get Amino acid tr...

Embodiment 2

[0022] Embodiment 2: the establishment of detection method

[0023] DNA extraction

[0024] Use Beijing Zhuangmeng International Biogene Technology Co., Ltd. Bacterial Genomic DNA Small Extraction Kit for extraction and recovery, the steps are as follows:

[0025] (1) Take 5 mL of bacterial culture solution, centrifuge at 12,000 rpm for 1 minute, and try to suck up the supernatant;

[0026] (2) Add 500 μL of cell suspension to the centrifuge tube with the bacterial pellet left, use a pipette or a vortex shaker to thoroughly suspend the bacterial cell pellet, and incubate at 37°C for 30 minutes. Mix by inversion several times every 10 minutes. Centrifuge at 12,000rpm for 2 minutes, and try to suck up the supernatant;

[0027] (3) Add 225 μL of buffer A to the cell pellet, shake until the cell is completely suspended;

[0028] (4) Add 6 μL RNaseA solution to the tube, shake for 15 seconds, and place at room temperature for 5 minutes;

[0029] (5) Add 10 μL of proteinase...

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Abstract

The invention discloses application of specific genes of staphylococcus epidermidis. The genes are Amino acid transporter LysE genes, and the Genbank login number is 3240523. Through the primer, whether the Amino acid transporter LysE genes exist in a sample or not is detected through a polymerase chain reaction, and then whether the staphylococcus epidermidis exists in the sample or not is determined. The staphylococcus epidermidis can be detected through the specific genes fast, easily, conveniently and accurately.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a new application of a specific gene of staphylococcus epidermidis, in particular applying the gene to establishing a technical method for detecting staphylococcus epidermidis. Background technique [0002] Staphylococcus epidermidis is an opportunistic pathogenic bacterium that exists on the surface of the skin, causing sepsis, and the tendency of chronic prostatitis is increasing day by day. The drug resistance of Staphylococcus epidermidis is very serious, and it is difficult to treat diseases caused by Staphylococcus epidermidis clinically. Therefore, it is particularly important to quickly detect Staphylococcus epidermidis. [0003] At present, the detection methods of staphylococcus epidermidis in clinical application include conventional smear microscopy, culture method, immunological determination, instrument automatic analysis and identification system, etc., but these methods...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/14
Inventor 宋玉竹刘有福张阿梅毛小琴夏雪山牛华孔令聘谢丽娇
Owner KUNMING UNIV OF SCI & TECH
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