Kit for digital PCR detection of Burkholderia

A technology for detection reagents and kits, applied in the field of digital PCR detection kits for Burkholderia, can solve the problems of inconvenient treatment, high detection sensitivity, long consumption time, etc., and achieve the effect of good sensitivity

Pending Publication Date: 2021-12-07
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI +1
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  • Abstract
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  • Application Information

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Problems solved by technology

In the prior art, the identification of Burkholderia is still mostly carried out by means of strain isolation and culture, but most of the methods of isolation and culture require 18-24 hours, which consumes a long time and brings inconvenience to the treatment
[0004] Digital PCR technology is an absolute quantitative technology for nucleic acid molecules. Its detection requires a short period of time, and the sample demand is low, and its detection sensitivity is high. If digital PCR technology can be applied to the detection of Burkholderia, it can greatly improve Detection efficiency, more timely determination of treatment options for infected patients, but there is no report on the application of digital PCR in the detection of Burkholderia

Method used

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  • Kit for digital PCR detection of Burkholderia
  • Kit for digital PCR detection of Burkholderia
  • Kit for digital PCR detection of Burkholderia

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Embodiment 1

[0040] 1. Primer probe design

[0041] 1.1 According to the sequence analysis results of Burkholderia cepacia, Burkholderia mallei, and Burkholderia pseudomallei, primers and probes were designed using Primer Express software; The primer probe of the source internal reference, the specific information is as follows:

[0042]

[0043] 2. Sample

[0044] 2.1 Bacterial DNA samples: Outsourced inactivated bacteria, genomic DNA was extracted by magnetic bead method.

[0045] 2.2 Human DNA samples: buccal swabs, genomic DNA extracted by magnetic bead method.

[0046] 2.3 Exogenous internal reference template: Synthesized by a biological company, dissolved and diluted for later use.

[0047] 3. Detection method

[0048] 3.1 Digital PCR amplification system

[0049] 3.1.1 25XB primer probe mixture

[0050] components stock solution (μM) Volume (μL / person) 50 x ROX dye 0.3 B-F 400 0.0375 B-R 400 0.0375 Bc-P1 100 0.0375 Bc-P2 100 ...

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Abstract

The invention relates to the technical field of biology, in particular to a kit for digital PCR detection of Burkholderia. The invention provides a primer probe group for digital PCR (Polymerase Chain Reaction) detection of Burkholderia, and provides a kit for digital PCR detection of Burkholderia. The primer and the probe group can be used for accurately identifying whether a sample contains DNA (Deoxyribose Nucleic Acid) of burkholderia cepacia, burkholderia pseudomallei and burkholderia pseudomallei or not, and the primer and the probe group have good sensitivity in detection of the three kinds of the burkholderia cepacia, the burkholderia pseudomallei and the burkholderia pseudomallei.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a digital PCR detection kit for Burkholderia. Background technique [0002] The genus Burkholderia (Burkholderia) belongs to the rRNA group I of Pseudomonas, and the species in the genus include Burkholderia cepacia (B.cepacia), Burkholderia cocos (B.cocovenenans), Burkholderia mallei (B.rnallei), Burkholderia pseudomallei (B.pseudomallei), Burkholderia caryophylla (B.caruophylli) , Burkholderia gladiolus (B.gladioli), Burkholderia pickeri (B.picketti) and Burkholderia solanaceae (B.solanacearum), etc. [0003] Species within the genus Burkholderia are infective to humans, animals, and plants, respectively, and are pathogenic bacteria. Among them, five species are related to human diseases, namely Burkholderia cepacia, Burkholderia mallei, Burkholderia pseudomallei, Burkholderia gladiolus Deutschia and Burkholderia Pieteri. Of these, Burkholderia cepacia infections have been most...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6851C12R1/01
CPCC12Q1/689C12Q1/6851C12Q2600/166C12Q2531/113C12Q2545/113C12Q2563/107
Inventor 杨启文杜娟夏江朱海涛
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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