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A method and kit for detecting multiple highly pathogenic pathogenic bacteria

A highly pathogenic and pathogenic bacteria technology, applied in the field of molecular biology and nucleic acid detection, can solve the problems of poor repeatability, tediousness, lack of highly pathogenic pathogens, etc.

Active Publication Date: 2016-12-14
SHANGHAI TELLGEN LIFE SCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The second is to detect and identify the DNA of the specific gene of the bacteria. This method does not require cultivation, which greatly shortens the detection time, and generally only takes a few hours to complete; but this method has a disadvantage, that is, each detection It can only judge whether it is a certain germ or not, and then conduct another reaction to judge whether it is another germ. Therefore, this method has a corresponding detection range, that is, it can only detect the germs within its detection range. If you want to detect Ten kinds of germs need to carry out ten reactions. Although these reactions can be carried out sequentially or in parallel, it is generally cumbersome.
However, like other solid-state chip detection, due to the defects of the solid-state chip itself, this method requires multiple washings, the process is still cumbersome, and the sensitivity is generally not high. More importantly, the repeatability is poor and the detection quality is unstable.
[0009] In summary, there is still a lack of satisfactory technology in the field that can simultaneously detect multiple highly pathogenic pathogenic bacteria

Method used

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  • A method and kit for detecting multiple highly pathogenic pathogenic bacteria
  • A method and kit for detecting multiple highly pathogenic pathogenic bacteria
  • A method and kit for detecting multiple highly pathogenic pathogenic bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0120] The detection of embodiment 1 highly pathogenic pathogenic bacteria

[0121] 1. Coating of probe microspheres:

[0122] 1. Synthesize probes according to the list below

[0123]

[0124] Note: The spacer arm is poly(T) composed of 10 Ts 10 -, so the probe Ban-P1 is equal to poly(T) 10 +SEQ ID NO.: 3, probe Ype-P1 is equal to poly(T) 10 +SEQ ID NO.: 4, and so on.

[0125] 2. Select 11 kinds of fluorescently encoded microspheres with numbers 22, 24, 26, 28, 31, 32, 33, 34, 35, 36 and 42 [Luminex Company], and carry out the probe corresponding to each probe in the above table in turn. Coated, the method is as follows:

[0126] (1) Equilibrate various microspheres and a portion of EDC powder stored at -20°C at room temperature for 30 minutes;

[0127] (2) Take each corresponding probe and dissolve it in double distilled water, the concentration is 0.01mM (10pmol / μL);

[0128] (3) Mix the microspheres evenly with an oscillator;

[0129] (4) Take 50ul each (6.0×10 ...

Embodiment 2

[0195] 1. Coating of probe microspheres:

[0196] 1. Synthesize probes according to the list below

[0197]

[0198] NOTE: The spacer arm is -HEG-.

[0199] 2. Select 11 kinds of fluorescently encoded microspheres with numbers 22, 24, 26, 28, 31, 32, 33, 34, 35, 36 and 42 [Luminex Company], and carry out the probe corresponding to each probe in the above table in turn. Coated, the method is as follows:

[0200] (1) Equilibrate various microspheres and a portion of EDC powder stored at -20°C at room temperature for 30 minutes;

[0201] (2) Take each corresponding probe and dissolve it in double distilled water, the concentration is 0.01mM (10pmol / μL);

[0202] (3) Mix the microspheres evenly with an oscillator;

[0203] (4) Take 50μl each (6.0×10 5 ) Microspheres, put into a clean 1.5ml centrifuge tube with pre-marked numbers;

[0204] (5) Centrifuge under the condition of 8000g centrifugal force for 5 minutes to precipitate the microspheres, carefully discard the supe...

Embodiment 3

[0265] The detection of embodiment 3 highly pathogenic pathogenic bacteria

[0266] Example 2 was repeated except that samples 16-27 were replaced with samples 28 and 29. Among them, samples No. 1 and No. 16 were combined as sample No. 28. Combine samples No. 3 and No. 25 as sample No. 29.

[0267] The test results showed that sample No. 28 contained Clostridium botulinum and Streptococcus suis; sample No. 29 contained tularafil and Legionella pneumophila. The test results are consistent with the actual situation of the samples.

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Abstract

The invention provides a method and kit for detecting multiple highly pathogenic pathogenic bacteria. Specifically, the present invention discloses a method for simultaneously detecting a variety of highly pathogenic pathogenic bacteria, including performing a PCR reaction with a specific primer set for highly pathogenic bacteria in a polymerase reaction system, thereby obtaining amplified amplification products; and detection with specific probes or probe microspheres. The invention also provides corresponding kits. The invention can detect and identify a variety of highly pathogenic bacteria sensitively and easily, including: Bacillus anthracis, Yarrowia pestis, Clostridium botulinum, Brucella, Streptococcus suis, Vibrio cholerae, Tula Vermicelli, Pseudomonas mallei (or Pseudomonas pseudomallei), Benacoxia, Legionella pneumophila.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and nucleic acid detection. Specifically, the present invention relates to a method and a kit for detecting a variety of highly pathogenic pathogenic bacteria. Background technique [0002] The pathogens that cause sudden infectious diseases can cause outbreaks of severe infectious diseases in peacetime, and can be used as biological weapons in wartime. These pathogenic microorganisms are mainly divided into six categories: viruses, bacteria, rickettsia, chlamydia, mycoplasma and fungi, and there are more than 40 kinds of them. Bacillus anthracis, Yersinia pestis, Clostridium botulinum, Brucella mallei, Brucella abortus, Streptococcus suis 2 suis II), Vibriocholerae, Francisella tularensis, Burkholderia mallei, Burkholderia pseudomallei, Coxiella burnetii ), Legionella pneumophila (Legionella pneumophila) etc. are highly pathogenic pathogenic bacteria. [0003] At present, the bacte...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/14C12Q1/04C12N15/11
Inventor 谭畅王勤熙张秀斐姚见儿
Owner SHANGHAI TELLGEN LIFE SCI CO LTD
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