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Primer for detecting chaetomium globosum in jet fuel, application and detection method

A technology of Chaetomium globosa and detection method, which is applied in the field of application and detection, detection of Chaetomium globosa primers in jet fuel, can solve the problems that the sensitivity and specificity cannot meet the needs of growing development, etc., to ensure the safety of aircraft flight, High sensitivity and good specific effect

Active Publication Date: 2020-06-19
LOGISTICAL ENGINEERING UNIVERSITY OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The current detection method is mainly PCR technology, but the sensitivity and specificity of this method cannot meet the growing needs

Method used

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  • Primer for detecting chaetomium globosum in jet fuel, application and detection method
  • Primer for detecting chaetomium globosum in jet fuel, application and detection method
  • Primer for detecting chaetomium globosum in jet fuel, application and detection method

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0041] Example 1: Primer Design and Synthesis

[0042] A set of specific primers were designed according to the β protein gene sequence of Chaetomium globosum, including outer primers (F3, B3), inner primers (FIP, BIP), and a loop primer (LB). The 5' end of FIP was biotinylated. The primer sequences are shown in Table 1,

[0043] Table 1 Primers for Chaetomium globosum

[0044]

[0045] According to the primer sequences in Table 1, LAMP primers were synthesized in Dalian Bao Biology Co., Ltd.

example 2

[0046] Example 2: Optimization of reaction conditions for amplification of Chaetomium globosum

[0047] The amplification conditions were optimized using the genomic DNA of Chaetomium globosum as a template. The concentrations of primers prepared during LAMP reaction were: 0.2 μmol / L for each of F3 and B3, 1.6 μmol / L for each of FIP and BIP, and 0.8 μmol / L for each of LB. LAMP premix is ​​20mmol / LTris-HCl (pH8.8), 10mmol / LKCl, 8mmol / L MgSO 4 , 10mmol / L (NH4) 2 SO 4 , 0.1% Tween 20, 0.8mol / L Betaine, 1.4mmol / L dNTPs, 8U / μL Bst DNA polymerase. The LAMP reaction system of Chaetomium globosum is shown in Table 2.

[0048] Table 2 LAMP reaction system of Chaetomium globosum

[0049]

[0050] In order to obtain the best LAMP amplification efficiency and comprehensively consider the applicable conditions of LFD, the primers provided by the present invention and the LAMP reaction system were used to perform LAMP amplification reactions at three different temperatures (61°C, 63...

example 3

[0052] Example 3 The LAMP-LFD technique of the present invention is used to determine the sensitivity of Chaetomium globosum.

[0053] 1. Using the extracted Chaetomium globosum DNA as a template, dilute to 10 times according to 10-fold equal gradient -8 Gradient (concentrations are 335, 33.5, 3.35ng / μL, 335, 33.5, 3.35pg / μL, 335, 33.5, 3.35fg / μL). Different dilutions of DNA were used as PCR, LAMP and LAMP-LFD reaction templates.

[0054] 2. Use LAMP, LAMP-LFD and PCR methods to detect Chaetomium globosum, and compare the sensitivity of the three methods.

[0055] 2.1 Amplify with the above-mentioned Chaetomium globosum (Chaetomium globosum) genomic DNA through gradient dilution as the template of the LAMP reaction, according to the primers in Example 1 and the reaction system and temperature and time in Example 2, use the LFD method provided by the invention Analyze the LAMP reaction product, see the results figure 1 .

[0056] 2.2 Amplify the above-mentioned Chaetomium g...

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Abstract

The invention belongs to the technical field of biological detection, and discloses a primer for detecting chaetomium globosum in jet fuel, application and a detection method. The primer for detectingthe chaetomium globosum comprises an outer primer F3, an outer primer B3, an inner primer FIP, an inner primer BIP and a loop primer LB; and the sequence of the outer primer F3 is shown as SEQ ID NO.1, the sequence of the outer primer B3 is shown as SEQ ID NO.2, the sequence of the inner primer FIP is shown as SEQ ID NO.3, the sequence of the inner primer BIP is shown as SEQ ID NO.4, and the sequence of the loop primer LB is shown as SEQ ID NO. 5. The detection method has the advantages of being high in sensitivity and specificity.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a primer for detecting Chaetomium globosa in jet fuel, an application and a detection method. Background technique [0002] The description of the background technology of the present invention belongs to the relevant technology related to the present invention, and is only used to illustrate and facilitate the understanding of the content of the present invention. prior art on the filing date. [0003] Microbial contamination of aviation fuel can lead to fuel quality degradation, accelerated corrosion of pipelines and storage tanks, clogged filters and threats to flight safety. The microorganisms in jet fuel not only affect the cleanliness and storage stability of the fuel, but also bring harm to the safety of equipment use, which has become a major problem that plagues the quality management of aviation fuel in my country. A next-generation Illumina MiSeq high-th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6844C12Q1/04C12N15/11C12R1/645
CPCC12Q1/6895C12Q1/6844C12Q2531/119C12Q2565/625
Inventor 苏鹏熊云李泽振范林君牛明明孙新枫朱鹏
Owner LOGISTICAL ENGINEERING UNIVERSITY OF PLA
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