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Candida albicans detection kit based on RPA rapid detection technology

A technology of Candida albicans and detection kit, which is applied in the direction of recombinant DNA technology, DNA/RNA fragments, microbial determination/inspection, etc., to meet the detection needs, reliable detection results, and low reaction temperature

Pending Publication Date: 2020-07-31
CHENGDU HAIZHIYUAN BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] As far as we know, there is currently no detection kit for Candida albicans developed based on RPA at home and abroad.

Method used

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  • Candida albicans detection kit based on RPA rapid detection technology
  • Candida albicans detection kit based on RPA rapid detection technology
  • Candida albicans detection kit based on RPA rapid detection technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1 Real-time fluorescence detection kit and its application method based on RPA constant temperature detection of Candida albicans

[0064] 1. Composition of the kit

[0065] 1.1 Primers and probes

[0066] The primer and probe sequences of the present invention are as shown in Table 1. In the probe sequence, the base substitution dSpacer (for exonuclease III recognition and cutting) between the 30th and 31st bases is tetrahydrofuran, and the 3' end is blocked. C3-spacer modification of the breaking group (it can also be phosphate biotin or amine); the thymine at the 29th base is replaced by the fluorescent group FAM (dT-FAM), and the thymine at the 32nd base is quenched Group BHQ1 substitution (dT-BHQ1).

[0067] Table 1 Primer and probe sequences

[0068]

[0069]

[0070] 1.2 Other reagents

[0071] 100 μl of 50× core enzyme mixture freeze-dried powder mixture, including recombinase, DNA polymerase, single-strand binding protein, and exonuclease I...

Embodiment 2

[0106] Example 2 The test strip detection kit and its application method based on RPA constant temperature detection of Candida albicans

[0107] 1. Composition of the kit

[0108] 1.1 Primers and probes

[0109] The primer and probe sequences are as shown in Table 4, and the 5' end of the downstream primer has biotin (biotin) label; In the probe sequence, the base substitution dSpacer (for endonuclease IV recognition cleavage) is tetrahydrofuran, and the 3' end is modified by a blocking group C3-spacer (it can also be phosphate biotin or amine).

[0110] Table 4 Primer and probe sequences

[0111]

[0112] 1.2 Other reagents

[0113] 100 μl 50×core enzyme mixture lyophilized powder mixture: recombinase, polymerase, single-chain binding protein, endonuclease IV (mass ratio 4:1:9:3).

[0114] 3ml rehydration buffer [2×Reaction Buffer (Tiwist DX TM )].

[0115] 500ul 280mM magnesium acetate.

[0116] Colloidal gold immunochromatographic test strips Consists of sample...

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PUM

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Abstract

The invention discloses a rapid, simple, specific and sensitive detection kit suitable for most laboratories for detecting Candida albicans based on RPA (recombinase polymerase amplification). The kitcomprises a pair of primers and a probe, the nucleotide sequences of the primers are shown as SEQIDNO.1-2, and the nucleotide sequence of the probe is shown as SEQIDNO.3. The kit has two forms: a real-time fluorescence detection kit and a test strip kit. Experiments show that the kit is short in candida albicans detection time and high in sensitivity and specificity, the cost is further saved bycombining a simple nucleic acid crude extraction method, a new technical reference can be provided for field and nursing point detection, and the kit is of great significance in the aspect of pathogenic bacteria detection in areas lack of resource basic equipment.

Description

technical field [0001] In the field of pathogenic microorganism inspection, it involves kits and applications for detecting Candida albicans, especially fluorescence detection kits, test strip detection kits and applications for rapid detection of Candida albicans based on RPA. Background technique [0002] Candida albicans is the most common fungus that causes invasive candidiasis (Invasive Candidiasis, IC), and is an important opportunistic pathogen. Under normal circumstances, it exists in the pharyngeal cavity, upper respiratory tract, intestinal tract and vagina of a normal population, and generally occurs in a small amount in a normal body and does not cause disease. When some risk factors (such as damage to the natural barrier of the skin and gastrointestinal tract, repeated or long-term use of broad-spectrum antibiotics, immune system damage, etc.) sick. Invasive candidiasis ranges from initial symptomatic candidemia to fulminant sepsis, with a mortality rate of ov...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6844C12Q1/04C12N15/11
CPCC12Q1/6844C12Q1/6895C12Q2600/166C12Q2531/119C12Q2563/107C12Q2521/507C12Q2521/101C12Q2522/101C12Q2545/113
Inventor 李晓鲁蓝海蒙雨丹
Owner CHENGDU HAIZHIYUAN BIOTECHNOLOGY CO LTD
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