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Method for detecting salmonella through PCR-DHPLC

A Salmonella and strain technology, applied in the field of bioanalytical chemistry, can solve the problems of low detection sensitivity, complicated and cumbersome inspection procedures, and long time-consuming, and achieve the effects of high sensitivity, simple result analysis, and improved sensitivity.

Inactive Publication Date: 2010-12-22
JIANGNAN UNIV
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Problems solved by technology

[0003] Traditional methods for detecting bacteria include biochemical culture, isolation and identification, the inspection procedures are complex and cumbersome, and it takes about 4-7 days to report the inspection results, which takes too long and the detection sensitivity is low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] (1) Enrichment culture of Salmonella:

[0052] A loop of Salmonella strain was inoculated in 10 mL of selenite cystine enrichment solution (SC enrichment solution), and cultured at 36° C. for 10 h. Salmonella (Salmonella), provided by Jiangsu Entry-Exit Inspection and Quarantine Bureau.

[0053] (2) Preparation of magnetic nanoparticles:

[0054] ① Take 0.65g FeCl 3 , 0.4g trisodium citrate, 20mL ethylene glycol, stirred for 10min.

[0055] ② Add 1.2g of sodium acetate to the above solution, and stir for 20 minutes.

[0056] ③The above solution was stirred and reacted at 230°C for 10h.

[0057] ④ Add 20 mL of absolute ethanol to the above solution, sonicate for 10 min, centrifuge at 6000 rpm for 10 min, and remove the supernatant.

[0058] ⑤ Add 20 mL of deionized water to the above precipitate, sonicate for 10 min, centrifuge at 6000 rpm for 10 min, and remove the supernatant.

[0059] ⑥ Add 5 mL of deionized water to the above precipitate.

[0060] (3) Extracti...

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PUM

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Abstract

The invention relates to a method for detecting salmonella through PCR-DHPLC, belonging to the technical field of bioanalytical chemistry. The method adopts PCR-DHPLC to detect salmonella and comprises the following steps: cultivating salmonella, preparing magnetic nanoparticles, extracting the DNAs of the salmonella genome, selecting the specific primer of salmonella, performing PCR amplification and performing DHPLC detection to the PCR product. The method of the invention adopts DHPLC instead of the traditional agarose electrophoresis to detect the PCR product, thus simplifying the reaction conditions, increasing the detection sensitivity, simplifying the operation and achieving the aim of large flux; and compared with the traditional microbial detection method and the PCR-gel electrophoresis method, the PCR-DHPLC detection method has the advantages that the sensitivity is high, the specificity is high, the detection is fast and accurate, the analysis of result is simple, the demand on samples is not high, the large flux detection can be performed, etc.

Description

technical field [0001] The invention discloses a method for detecting Salmonella by using PCR-DHPLC, which belongs to the technical field of bioanalytical chemistry. Background technique [0002] Pathogenic microorganisms are widely distributed in nature, can invade human body and animals, cause infection and even infectious diseases, and pose great harm to human and animal health. Prevention of pathogenic microorganisms is of great significance in public health. Eating food contaminated with pathogenic bacteria is the main route of transmission of foodborne diseases. Human and animal infection can be asymptomatic or clinically symptomatic, which may aggravate morbidity or mortality, or reduce the reproductive capacity of animals. [0003] The traditional methods for detecting bacteria include biochemical culture, isolation and identification, the inspection procedures are complex and cumbersome, and it takes about 4-7 days to report the inspection results, which is too tim...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N30/02C12R1/42
Inventor 胥传来勇倩倩陈伟刘微波徐丽广赵书阁
Owner JIANGNAN UNIV
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