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Targeted database creation kit for plurality of pathogens

A kit and pathogen technology, applied in DNA/RNA fragmentation, microbial determination/inspection, recombinant DNA technology, etc., can solve the problems of high noise background, long cycle, low throughput, etc., and achieve narrow identification range and rapid identification Effect

Active Publication Date: 2018-09-11
THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

The detection and detection of pathogens is an important part of the prevention and control of infectious diseases. In the face of thousands of known and unknown pathogens, the traditional isolation and cultivation of pathogens has blindness, low throughput, and long cycle. The confirmation of pathogens requires epidemiology, pathogenic However, conventional metagenomic next-generation sequencing has disadvantages such as high noise background, time-consuming and laborious analysis, etc.

Method used

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  • Targeted database creation kit for plurality of pathogens
  • Targeted database creation kit for plurality of pathogens
  • Targeted database creation kit for plurality of pathogens

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0010] Pathogen Panel establishment and verification

[0011] 43 pairs of multiplex PCR primers and corresponding positive templates were artificially synthesized, and the practicability and feasibility of the kit were preliminarily verified by next-generation sequencing analysis.

[0012] (1) Specific primer sequence design and synthesis

[0013] (2) Multiplex PCR reaction system establishment and positive template synthesis

[0014] See Table 1 for a list of primers and the lengths of synthetic template fragments.

[0015] Table 1. 43 kinds of pathogen primer sequence design

[0016]

[0017]

[0018]

[0019] a. Establishment of multiplex PCR system

[0020] The reaction system preparation and reaction conditions are as follows:

[0021] Multiplex PCR reaction system preparation

[0022]

[0023] Multiplex PCR reaction conditions: pre-denaturation at 95°C for 10 min; denaturation at 98°C for 15 sec, annealing at 60°C for 5 min, 10 cycles; storage at 10°C. ...

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Abstract

The invention relates to a targeted database creation kit for 43 pathogens. The pathogens comprise bacteria, viruses, fungi and protozoa. Targeted detection primer sequences of the pathogens are shownas SEQ ID NO. 1 to 86. According to the kit, targeted database creation can be performed on pathogen nucleic acids in an early infection stage for rapid diagnosis of the pathogens after next-generation sequencing, and directions are provided for subsequent pathogen isolation and identification, serologic detection, clinical diagnosis and symptomatic treatment.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a pathogen targeting library construction kit and a multiplex PCR system. Background technique [0002] With the development of society and economy, the threat of emerging infectious diseases to global public health security is increasing day by day. The detection and detection of pathogens is an important part of the prevention and control of infectious diseases. In the face of thousands of known and unknown pathogens, the traditional isolation and cultivation of pathogens has blindness, low throughput, and long cycle. The confirmation of pathogens requires epidemiology, pathogenic However, conventional metagenomic next-generation sequencing has disadvantages such as high noise background and time-consuming and laborious analysis. If rapid targeted amplification of pathogenic nucleic acid is performed in the early stage of infectious diseases, followed by high-throughput sequencing a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12Q1/686C12N15/11
CPCC12Q1/6806C12Q1/686C12Q2537/143
Inventor 邓国宏郑钧丰孙凤明谭文婷王修华
Owner THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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