Staphylococcus aureus PCR (polymerase chain reaction) detection primer, kit and detection method

A detection kit and technology for Staphylococcus, which is applied in the field of PCR detection primers for Staphylococcus aureus, can solve the problem of less application of cosmetic samples, and achieve the effects of saving detection cost and time, wide market application prospects, and high sensitivity

Inactive Publication Date: 2016-11-09
GUANGDONG INST OF MICROORGANISM
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of these methods focus on drug resistance-associated genes and toxin genes, and are rarely applied to the detection of cosmetic samples

Method used

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  • Staphylococcus aureus PCR (polymerase chain reaction) detection primer, kit and detection method
  • Staphylococcus aureus PCR (polymerase chain reaction) detection primer, kit and detection method
  • Staphylococcus aureus PCR (polymerase chain reaction) detection primer, kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Screening and determination of multiplex PCR primers for detection of Staphylococcus aureus

[0038]1. Primer design

[0039] Candidate detection target genes or sequences for the detection of Staphylococcus aureus were screened by consulting relevant references, and then their nucleotide sequences were analyzed by bioinformatics. After analysis, the thermostable nuclease gene (nuc gene) was selected in this study , fibrinogen binding protein gene (clf A gene) and specific sequence (SmaI sequence) as specific detection target sequence.

[0040] Primers were designed for the above-mentioned detection target sequences, and then through experimental verification, 3 pairs of primers with high specificity were screened out as follows:

[0041] Primer pair for thermostable nuclease gene (nuc gene) amplification:

[0042] nuc-F: 5'-GGGATGGCTATCAGTAA-3' (as shown in SEQ ID NO.1),

[0043] nuc-R: 5'-TGAATCAGCGTTGTCTT-3' (as shown in SEQ ID NO.2);

[0044] Primer p...

Embodiment 2

[0054] Example 2: Detection of Staphylococcus aureus in cosmetics

[0055] Six different types of cosmetic samples sent by a company for inspection, namely moisturizing lotion, lotion, face cream, facial cleanser, toner and liquid foundation, were tested using the microbial detection method in the "Hygienic Standards for Cosmetics" (2007 edition), and none of them were found Microbial contamination, artificial random pathogen contamination, add Staphylococcus aureus ATCC 6538 to moisturizer, add Staphylococcus aureus ATCC 6538 and Escherichia coli ATCC 8039 to lotion, add Staphylococcus aureus ATCC6538 and Pseudomonas aeruginosa to face cream coli ATCC 9027, Escherichia coli ATCC 8039 was added to cleansing cream, Pseudomonas aeruginosa ATCC 9027 was added to toner, Escherichia coli ATCC 8039 and Pseudomonas aeruginosa ATCC 9027 were added to foundation liquid, and each The final concentration of the bacterial suspension of the sample was 10 2 cfu / mL, thus preparing artificia...

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Abstract

The invention discloses a staphylococcus aureus PCR (polymerase chain reaction) detection primer, a kit and a detection method. Through the combined utilization of a staphylococcus aureus heat-resistant nucleic acid enzyme gene (nuc gene), a fibrinogen binding protein gene (clf A gene) and a specificity sequence, the staphylococcus aureus PCR is specifically designed, and the staphylococcus aureus multiple PCR detection primer, the kit and the detection method are developed; the technical effects of strong detection specificity and high sensitivity are realized, the detection consumes less time, the operation is convenient and simple, the detection efficiency can be greatly increased, the requirement of rapid detection is met, and the staphylococcus aureus PCR (polymerase chain reaction) detection primer, the kit and the detection method can be applied to practical production work.

Description

technical field [0001] The invention belongs to the field of microorganism detection, and in particular relates to a PCR detection primer, a kit and a detection method for Staphylococcus aureus. Background technique [0002] With the improvement of human living standards, cosmetics have gradually become a necessity in people's lives. Various nutritional elements in cosmetics provide a good living environment for microorganisms. Once the microorganisms are contaminated in cosmetics, it is bound to endanger people's health, especially the golden yellow grapes. Coccus pollution, Staphylococcus aureus is the most pathogenic to humans in the Staphylococcus genus, can cause local purulent lesions in the human body, and in severe cases can lead to sepsis, it can not only infect the respiratory tract, poison the digestive tract, but also Drilling into the skin induces dermatitis. Therefore, Staphylococcus aureus is listed as a routine inspection item for cosmetics in my country's "Hyg...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/14C12N15/11
CPCC12Q1/686C12Q1/689C12Q2600/16C12Q2537/143C12Q2527/125
Inventor 文霞杨秀茳谢小保孙廷丽李彩玲
Owner GUANGDONG INST OF MICROORGANISM
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