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Rapid detection primers, kit and detection method for C.parapsilosis with loop-mediated isothermal amplification

A technology of Candida parapsilosis and ring-mediated isothermal, which is applied in biochemical equipment and methods, microbial measurement/testing, DNA/RNA fragments, etc., to achieve high sensitivity, rapid detection, and strong specificity

Inactive Publication Date: 2013-02-20
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the LAMP detection method for Candida parapsilosis

Method used

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  • Rapid detection primers, kit and detection method for C.parapsilosis with loop-mediated isothermal amplification
  • Rapid detection primers, kit and detection method for C.parapsilosis with loop-mediated isothermal amplification

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Embodiment Construction

[0012] The key of the invention lies in the selection of target gene and the design of primers. The inventors of the present invention selected a special sequence in the conserved sequence of Candida parapsilosis as the target gene, and determined that it is unique to Candida parapsilosis after searching and comparing in GenBank. For this sequence, the primer explorer V3 software was used to design primers. The software gives 1000 sets of candidate primers. Finally, the following 2 pairs of primers were obtained, and the primer sequences were:

[0013] SEQ ID NO 1:

[0014] 5'-GCCTACCAAAAGCAAAGTTTTTCAAAA-GAATGAAAAGTGCTTAACTGC-3';

[0015] SEQ ID NO 2:

[0016] 5'-GGCCTGCCAGAGATTAAACTC-CCGTTGTTGAAAGTTTTGAC-3';

[0017] SEQ ID NO 3:

[0018] 5'-TAGGTGAACCTGCGGAAG-3';

[0019] SEQ ID NO 4:

[0020] 5'-GATGCGAGAACCAAGAG-3'.

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Abstract

The invention provides rapid detection primers, a kit and a detection method for C.parapsilosis with loop-mediated isothermal amplification (LAMP). The LAMP rapid detection primers for C.parapsilosis comprise a pair of inner primers and a pair of outer primers, wherein, the sequences of the inner primers are shown as the SEQ ID No.1 and SEQ ID NO.2, the sequences of the outer primers are shown as the SEQ ID No.3 and SEQ ID NO.4. By utilizing the invention to detecting C.parapsilosis, the advantages of high sensitivity, strong singularity, rapid detection and simple operation are achieved.

Description

technical field [0001] The invention belongs to the detection technology scope of biological fungi, and in particular relates to rapid detection primers, a kit and a detection method for candida parapsilosis ring-mediated isothermal amplification. Background technique [0002] Loop-mediated isothermal amplification (LAMP for short) is to design 2 pairs of special primers for 6 specific sites on the target sequence, and through DNA polymerase with strand displacement activity, under constant temperature conditions A new technology for specifically, efficiently and rapidly amplifying DNA. The amplified product is a mixture of DNA fragments with a stem-loop structure and a polycyclic cauliflower-like structure composed of a series of inverted repeats of the target sequence. Existing technologies apply LAMP to the detection of cytomegalovirus, mycobacteria, salmonella, paracoccidioides brasiliensis, and African trypanosoma. The key to its specificity and efficiency lies in the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
Inventor 沈定霞王凯飞刘朝军叶丽艳郭玲张有江
Owner GENERAL HOSPITAL OF PLA