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Method of quickly identifying human fungi based on sanger sequencing

A sequencing and fungal technology, applied in the field of high-throughput sequencing, can solve the problems of easy confusion, not necessarily correct and reliable strains, and achieve high accuracy and avoid cross-contamination

Pending Publication Date: 2019-05-07
SHANGHAI PASSION BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Since the colony morphology of many bacteria is very similar, it is easy to cause confusion, and with the evolution of species, many bacteria are resistant to drugs, so the bacteria identified by conventional methods are not necessarily correct and reliable, and have defects.

Method used

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  • Method of quickly identifying human fungi based on sanger sequencing
  • Method of quickly identifying human fungi based on sanger sequencing
  • Method of quickly identifying human fungi based on sanger sequencing

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Experimental program
Comparison scheme
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Embodiment Construction

[0037] see Figure 1-6 , the specific embodiment of the present invention is:

[0038] 1. Extraction of bacterial or fungal chromosomal DNA (manufactured by Zhangjiagang Lansu Biological Engineering Co., Ltd.)

[0039] 1.1 Secretion (fungus) DNA extraction kit

[0040] 1.11 For samples stored at room temperature or at 2-8°C, proceed directly to the next step; for samples stored in a refrigerator below -20°C, place them at room temperature to completely melt the samples before proceeding to the next step.

[0041] 1.12 Take out all the suspension and put it into a 1.5ml centrifuge tube, centrifuge at 13000r / min for 3 minutes, discard the supernatant.

[0042] 1.13 Add 1ml of sample cleaning solution to shake and resuspend, centrifuge at 13000r / min for 3 minutes, and discard the supernatant.

[0043] 1.14 Add 300 μl of sample treatment solution A to resuspend the pellet. Transfer all the resuspended samples to centrifuge tubes containing DNA solid extracts (pre-centrifuge be...

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PUM

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Abstract

The invention discloses a method of quickly identifying human fungi based on sanger sequencing. The method is characterized by comprising the steps of DNA (deoxyribonucleic acid) extraction and quality inspection, primer design and synthesis, PCR (polymerase chain reaction) amplification, PCR product purification, PCR product purification, sequencing and data processing. The method has the benefits that a gene sequence of a particular species can be amplified through a specific primer, verification and comparison are performed through the sequencing, and an accuracy rate is almost close to 100% through one-to-one correspondence. DNA of certain colonies is only required to be extracted and stored, and misjudgment due to cross contamination in a culture process is avoided. The method is convenient, quick and high in accuracy as compared with other methods.

Description

technical field [0001] The invention belongs to the technical field of high-throughput sequencing, and in particular relates to a method for rapidly identifying human fungi based on Sanger sequencing. Background technique [0002] Several fungal detection methods commonly used today are: [0003] 1. Direct microscope detection Take the secretions and other materials to be tested and smear them on a glass slide, and observe the structure of mycelium under the microscope to determine what kind of bacteria it is. [0004] 2. Isolation and culture specimens were inoculated in a specific medium, and colonies were formed after incubation at room temperature. The bacteria were judged by their appearance, and the colonies were smeared for microscopic examination. [0005] 3. Drug sensitivity test [0006] Since the colony shapes of many bacteria are very similar, it is easy to cause confusion, and with the evolution of species, many bacteria are resistant to drugs, so the bacteria...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6869
CPCC12Q1/6895C12Q1/6869
Inventor 辛文斌孙子奎丁方美王锋
Owner SHANGHAI PASSION BIOTECHNOLOGY CO LTD