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Specific primer and kit for rapid detection of candida albicans

A Candida albicans and kit technology, applied in the directions of microorganisms, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of low fever, high fatality rate, complicated methods, etc., and achieve real-time detection, high-efficiency specific amplification, Amplify the effect of strong compatibility

Pending Publication Date: 2020-06-19
深圳华薇生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The diagnosis of invasive candidiasis is difficult. The early symptoms are usually persistent low-grade fever, and the traditional gold standard for identification, that is, the culture method, takes at least two days to obtain the experimental results, which often prevents patients from receiving timely treatment and leads to a high mortality rate.
Molecular biology techniques such as PCR, real-time fluorescent PCR, and immunoassays have also been used for the detection of Candida albicans, but these methods are relatively complicated

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Primer design:

[0026] The nucleotide sequences of the specific primers used to detect Candida albicans are as follows:

[0027] Ft: 5'-TCTTTTTCGATGCGTACTGGACCAGCCGAGCCTTTCCTTCTGGGTAGC-3';

[0028] Bt: 5'-TGGTTTCTAGGACCATCGTAATGATTAATAGGGACGGTCG-3';

[0029] IF: 5'-AATTTGACTC-3';

[0030] IB: 5'-CCTTGGCCGAGAGGTCTG-3'.

[0031] 2. Kit design:

[0032] The kit includes reagent one, reagent two and blocking agent;

[0033] Wherein, reagent one comprises primer Ft, Bt, primer IF, IB, Bst DNA polymerase, dNTP and indicator;

[0034] Reagent two comprises Tris-HCl, ammonium sulfate, magnesium sulfate, potassium chloride, Tween-20 and betaine;

[0035] The sealing agent is paraffin.

[0036] Among them, the final concentration of each reagent in the kit is: Ft, Bt each 1.5μM, IF, IB each 0.9μM, dNTP each 1.5mM, Bst DNA polymerase 8U, Tris-HCl 20.0mM, ammonium sulfate 10mM, potassium chloride 50mM , Magnesium Sulfate 10mM, Tween-200.1%, Betaine 0.7M, pH 8.5.

Embodiment 2

[0038] 1. Primer design:

[0039] The nucleotide sequences of the specific primers used to detect Candida albicans are as follows:

[0040] Ft: 5'-TCTTTTTCGATGCGTACTGGACCAGCCGAGCCTTTCCTTCTGGGTAGC-3';

[0041] Bt: 5'-TGGTTTCTAGGACCATCGTAATGATTAATAGGGACGGTCG-3';

[0042] IF: 5'-AATTTGACTC-3';

[0043] IB: 5'-CCTTGGCCGAGAGGTCTG-3'.

[0044] 2. Kit design:

[0045] The kit includes reagent one, reagent two and blocking agent;

[0046] Wherein, reagent one comprises primer Ft, Bt, primer IF, IB, Bst DNA polymerase, dNTP and indicator;

[0047] Reagent two comprises Tris-HCl, ammonium sulfate, magnesium sulfate, potassium chloride, Tween-20 and betaine;

[0048] The sealing agent is paraffin.

[0049] Among them, the final concentration of each reagent in the kit is: Ft, Bt each 1.2μM, IF, IB each 0.7μM, dNTP each 1.2mM, Bst DNA polymerase 6U, Tris-HCl 20.0mM, ammonium sulfate 10mM, potassium chloride 50mM , Magnesium Sulfate 10mM, Tween-200.1%, Betaine 0.7M, pH 9.

Embodiment 3

[0051] 1. Primer design:

[0052] The nucleotide sequences of the specific primers used to detect Candida albicans are as follows:

[0053] Ft: 5'-TCTTTTTCGATGCGTACTGGACCAGCCGAGCCTTTCCTTCTGGGTAGC-3';

[0054] Bt: 5'-TGGTTTCTAGGACCATCGTAATGATTAATAGGGACGGTCG-3';

[0055] IF: 5'-AATTTGACTC-3';

[0056]IB: 5'-CCTTGGCCGAGAGGTCTG-3'.

[0057] 2. Kit design:

[0058] The kit includes reagent one, reagent two and blocking agent;

[0059] Wherein, reagent one comprises primer Ft, Bt, primer IF, IB, Bst DNA polymerase, dNTP and indicator;

[0060] Reagent two comprises Tris-HCl, ammonium sulfate, magnesium sulfate, potassium chloride, Tween-20 and betaine;

[0061] The sealing agent is paraffin.

[0062] Among them, the final concentration of each reagent in the kit is: Ft, Bt each 1μM, IF, IB each 0.5μM, dNTP each 1mM, BstDNA polymerase 6U, Tris-HCl 20.0mM, ammonium sulfate 10mM, potassium chloride 50mM, magnesium sulfate 10mM, Tween-200.1%, Betaine 0.6M, pH 9.5.

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Abstract

The present invention relates to the technical field of biological detection and particularly to a specific primer and a kit for rapid detection of candida albicans. Firstly, the specific primer is provided, besides, the specific primer is relatively critical, and then the specific primer is used to prepare a buffer system to form the kit for the rapid detection of the candida albicans.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a specific primer and a kit for rapid detection of Candida albicans. Background technique [0002] Candida albicans (Candida albicans) is the most important human opportunistic pathogenic fungus. It usually exists on the surface of normal human skin and mucous membranes. Bacteria multiply and cause diseases, which can be summarized into two categories: mucocutaneous candidiasis and invasive or deep organ candidiasis. [0003] Mucocutaneous candidiasis mainly includes thrush, interdigital erosion, papuloid, paronychia and gynecological candidiasis. Among them, oral and vulvovaginal candidiasis is the most common, which is characterized by a white film of bacteria that is easily scraped off on the affected area. Oral candidiasis (also known as oral thrush) occurs mostly in infants and young children under 2 years old, and is less common in adults. Vulvovaginal candi...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/04C12N15/11C12R1/725
CPCC12Q1/6895
Inventor 张勇
Owner 深圳华薇生物科技有限公司
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