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PCR (Polymerase Chain Reaction) assay kit for rana boulenger guenther-infecting staphylococcus aureus and assay method

A detection kit and staphylococcus technology, applied in the fields of bioengineering technology and biomedicine, can solve problems such as lack of specificity, and achieve the effect of strong specificity and high sensitivity

Inactive Publication Date: 2015-12-23
CHONGQING UNIV OF ARTS & SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0015] The present invention finds that the PCR kit using the above primers is not sensitive and lacks specificity in detecting the infection of Staphylococcus aureus in spiny-bellied frogs

Method used

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  • PCR (Polymerase Chain Reaction) assay kit for rana boulenger guenther-infecting staphylococcus aureus and assay method
  • PCR (Polymerase Chain Reaction) assay kit for rana boulenger guenther-infecting staphylococcus aureus and assay method
  • PCR (Polymerase Chain Reaction) assay kit for rana boulenger guenther-infecting staphylococcus aureus and assay method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 4

[0096] Embodiment 4 PCR rapid detection kit of the present invention is to the detection of pathogenic spiny-bellied frog

[0097] Using the kit described in Example 1, proceed as follows:

[0098] (1) The DNA extracted from the tissues of 6 spiny-bellied frogs taken from the affected farms were used as templates for PCR detection.

[0099] (2) Take 10 μL of 2-fold reaction mixture buffer, 0.5 μL each of upstream and downstream primers (S-F, S-R), 1 μL of TaqDNA polymerase, ddH 2 07 μL, DNA template 1.0 μL, and another 1 μL of positive control solution and negative control solution were taken as templates, as positive and negative control groups. After mixing evenly, centrifuge for a few seconds and place on a PCR reaction instrument for reaction.

[0100] (3) Carry out PCR amplification reaction according to the following conditions: pre-denaturation at 95°C for 5 minutes; then denaturation at 95°C for 30 seconds, annealing at 56°C for 30 seconds, extension at 72°C for 30 s...

Embodiment 5

[0103] Example 5 Detection of the PCR rapid detection kit of the present invention to the samples of the spiny-bellied frog in different regions

[0104] Using the kit described in Example 1, proceed as follows:

[0105] (1) Using samples of Rana spinosa with symptoms of Staphylococcus aureus infection collected from Sichuan, Guizhou, Chongqing and other places, the DNA in the tissue was extracted as a template for PCR detection.

[0106] (2) Take 10 μL of 2-fold reaction mixture buffer, 0.5 μL each of upstream and downstream primers (S-F, S-R), 1 μL of TaqDNA polymerase, ddH 2 07 μL, template 1.0 μL, and take 1 μL each of the positive control solution and the negative control solution as the template, as the positive and negative control groups. After mixing evenly, centrifuge for a few seconds and place on a PCR reaction instrument for reaction.

[0107] (3) Carry out PCR amplification reaction according to the following conditions: pre-denaturation at 95°C for 5 minutes; ...

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Abstract

The invention discloses a PCR (Polymerase Chain Reaction) rapid assay kit for rana boulenger guenther-infecting staphylococcus aureus and an assay method. The PCR rapid assay kit comprises a PCR assay kit for assaying the staphylococcus aureus of ana boulenger guenthers, which includes 2X mixed reaction buffer, sense primer S-F:5minute -AGGTAACGGCGTAAATAG-3minute, antisense primer S-R: 5minute-ACTTGCTTCAGGACCATA-3minute, Taq enzyme and sterilized ddH2O. The PCR rapid assay kit disclosed by the invention has the advantages of rapidness, accuracy and specificity in the assay of the rana boulenger guenther-infecting staphylococcus aureus, meets the requirement of clinical diagnosis and provides convenient conditions for the assay of the staphylococcus aureus infecting the rana boulenger guenthers.

Description

technical field [0001] The invention belongs to the fields of bioengineering technology and biomedicine, and in particular relates to a PCR detection reagent and a detection method for staphylococcus aureus of the spiny-bellied frog, which are suitable for detecting the spiny-bellied frog infected with the staphylococcus aureus. Background technique [0002] The spiny-bellied frog is a large-scale edible frog with rich nutrition and medicinal value. The protein content of its fresh meat jerky sample is 16.25%, and the fat content is 1.26%. It has rich nourishing effects and medicinal value. With the continuous improvement of the level, the edible value of spiny-bellied frog is also gradually rising, and it has a good development prospect. At the same time, as a transitional animal in the evolution of vertebrates, it is an ideal experimental animal for the study of anatomy, genetics and embryonic development. Due to serious environmental pollution, destruction of wild habita...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/14
CPCC12Q1/686C12Q2545/113
Inventor 姜玉松徐敬明樊汶樵吴宝红李晓英
Owner CHONGQING UNIV OF ARTS & SCI
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