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Nucleic acid aptamer, complementary sequence and detection method for detecting hemolytic streptococcus

A technology of hemolytic streptococcus and nucleic acid aptamer, applied in the field of clinical testing, can solve the problems of expensive reagents, difficult to popularize, etc., and achieve the effects of large detection volume, direct and objective results, and simple operation steps

Inactive Publication Date: 2013-03-27
HUNAN FOOD SAFETY PRODN ENG TECH RES CENT
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Problems solved by technology

However, these modern detection methods for single group hemolytic streptococcus all need certain equipment and technical requirements, and the cost of reagents is relatively expensive, so it is difficult to popularize

Method used

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  • Nucleic acid aptamer, complementary sequence and detection method for detecting hemolytic streptococcus
  • Nucleic acid aptamer, complementary sequence and detection method for detecting hemolytic streptococcus
  • Nucleic acid aptamer, complementary sequence and detection method for detecting hemolytic streptococcus

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Embodiment 1

[0033] 1. Experimental materials

[0034] 1.1 Fluorescence spectrophotometer measurement (Hitachi, Japan F7000); transmission electron microscope (JEM-1230, JEOL, Japan); pH meter (Shanghai REX Instrument Factory)

[0035] 1.2 Culture medium: brain heart infusion broth and blood plate are produced by Guangdong Huankai Microbial Technology Co., Ltd.

[0036] 1.3 Species: Hemolytic Streptococcus [CMCC (B) 32210], Staphylococcus aureus [ATCC6538], Salmonella [CMCC (B) 50115], Pseudomonas aeruginosa [CMCC (B) 10104], Bacillus sakazakii [ ATCC29004].

[0037] 2. A nucleic acid aptamer sequence marked with FAM that can specifically bind to hemolytic streptococcus was synthesized; at the same time, a complementary sequence of a DNA target sequence was synthesized.

[0038] 3. Sample treatment: In the biological safety cabinet, the sample was diluted and enriched according to the requirements of GB4789.11-2003, and the sample enrichment solution was incubated in a 37°C incubator for...

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Abstract

The invention relates to a nucleic acid aptamer, a complementary sequence and a nondiagnostic detection method for detecting hemolytic streptococcus, and belongs to the fields of clinical examination and food detection. The nucleic acid aptamer which is connected with an FAM fluorescence signal and is in specific combination with the hemolytic streptococcus, and an oligonucleotide sequence which is complementary with the aptamer are designed to establish a nucleic acid aptamer biosensor detection method for detecting the hemolytic streptococcus on the basis of colloidal gold quenching fluorescence. The nucleic acid aptamer, the complementary sequence and the detection method are mainly used for quickly detecting the hemolytic streptococcus, and have the advantages of strong specificity, high sensitivity, high detection efficiency, simplicity in operation and high safety.

Description

technical field [0001] The invention belongs to the field of clinical testing and food testing, and in particular relates to a nucleic acid aptamer, a complementary sequence and a method for detecting hemolytic streptococci based on the fluorescence quenching of colloidal gold. Background technique [0002] Hemolytic streptococci are spherical or elliptical, with a diameter of 0.6-1.0 μm, and are arranged in chains with different lengths, ranging from 4-8 to 20-30 bacterial cells. The length of the chain is related to the type and growth of the bacteria Environment related. The bacterium does not form spores, has no flagella, is easily stained by common basic dyes, is Gram-positive, and turns Gram-negative after aging culture or phagocytosis by neutrophils. [0003] The bacterium is an aerobic or facultative anaerobic bacterium with high nutritional requirements. It grows poorly on ordinary medium and needs to be supplemented with serum, blood, and ascites. Most strains nee...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/14C12Q1/06C12N15/115C12N15/11G01N21/64
Inventor 彭新凯李乐聂静苑钟菲菲刘子音黄亮杨丽霞夏立新
Owner HUNAN FOOD SAFETY PRODN ENG TECH RES CENT
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