Kit for detecting non-mucus type klebsiella pneumoniae with high virulence

A Klebsiella pneumoniae and kit technology are applied in the field of kits for detecting highly virulent non-mucus-type Klebsiella pneumoniae, and kits for detecting the virulence level of Klebsiella pneumoniae, and can solve the problem of inability to identify non-mucus type Klebsiella pneumoniae. The virulence of mucoid Klebsiella pneumoniae is easy to judge the results, the amplification efficiency and quality are improved, and the design principle is reasonable.

Pending Publication Date: 2020-01-10
SHENZHEN PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In this technical scheme, the kit is amp R The gene is a biomarker, and based on the polymerase chain reaction, the qualitative detection of highly virulent non-mucinous Klebsiella pneumoniae is completed to make up for the detection method of high-virulence Klebsiella pneumoniae in the prior art. The virulence of non-mucinous Klebsiella pneumoniae cannot be identified by the high mucus type

Method used

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  • Kit for detecting non-mucus type klebsiella pneumoniae with high virulence
  • Kit for detecting non-mucus type klebsiella pneumoniae with high virulence
  • Kit for detecting non-mucus type klebsiella pneumoniae with high virulence

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Experimental program
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Effect test

Embodiment 1

[0046] non-mucinous Klebsiella pneumoniae amp R Application of the gene as a marker in the preparation of a kit for detecting highly virulent non-mucinous Klebsiella pneumoniae, if the marker is tested positive, it is judged that the non-mucinous Klebsiella pneumoniae is highly virulent non-mucinous Klebsiella pneumoniae; on the contrary, if the marker is tested negative, it is judged that the non-mucinous Klebsiella pneumoniae is low-virulence non-mucinous Klebsiella pneumoniae;

[0047] said amp R The gene sequence is shown in SEQ ID NO.1.

Embodiment 2

[0049] A kit for detecting highly virulent non-mucinous Klebsiella pneumoniae, including a PCR reaction solution, primers for amplifying the marker SEQ ID NO.1, a positive control and a negative control; wherein,

[0050] PCR reaction solution: including Taq polymerase with a concentration of 5U / μL, tris hydrochloride solution with a concentration of 200mmol / L, magnesium chloride solution with a concentration of 20mmol / L, potassium chloride solution with a concentration of 500mmol / L, volume ratio It is 0.2% triton solution, the volume ratio is 10% formamide solution and the concentration is 10mmol / L deoxyribonucleoside triphosphate;

[0051] Primers: including upstream primers and downstream primers, the sequence of the upstream primer is 5'-AACAGGCGGCTGAACATACA-3', the sequence of the downstream primer is 5'-TTTACCCTGCTGCGCTCATT-3', and the concentrations of both the upstream primer and the downstream primer are 0.5 μmol / L;

[0052] Positive control: a non-mucinous Klebsiella...

Embodiment 3

[0055] On the basis of embodiment 1-2, further, such as figure 1 Shown:

[0056] The method for using the kit for detecting highly virulent non-mucinous Klebsiella pneumoniae comprises the following steps:

[0057] S1: Sampling

[0058] Inoculating the non-mucinous Klebsiella pneumoniae to be detected in the culture medium, culturing, and taking the culture fluid as a sample of the non-mucinous Klebsiella pneumoniae to be detected;

[0059] S2: PCR amplification system preparation

[0060] Take four PCR tubes and number them as positive control tubes, negative control tubes, experimental tubes and blank control tubes; the PCR amplification system is 20 µL, of which,

[0061] Add 17 μL PCR reaction solution, 1 μL upstream primer, 1 μL downstream primer and 1 μL positive control to the positive control tube;

[0062] Add 17 μL PCR reaction solution, 1 μL upstream primer, 1 μL downstream primer and 1 μL negative control to the negative control tube;

[0063] Add 17 μL of PC...

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Abstract

The invention discloses a kit for detecting non-mucus type klebsiella pneumoniae with high virulence, and belongs to the technical field of biological detection. An ampR gene sequence in a non-mucus type klebsiella pneumoniae genome with high virulence is used as a marker, primers are designed based on the specific sequence, and finally qualitative detection of the non-mucus type klebsiella pneumoniae with high virulence is completed based on a polymerase chain reaction to make up for the situation that a qualitative detection method of klebsiella pneumoniae in the prior art can determine thevirulence of the klebsiella pneumoniae only by judging whether a mucus type exists or not, but cannot distinguish the difference in virulence among non-mucus type strains, so that the clinical application value of the marks and kit for detecting the non-mucus type klebsiella pneumoniae with high virulence and an application method of the kit is ensured.

Description

technical field [0001] The invention relates to a kit for detecting highly virulent non-mucinous Klebsiella pneumoniae, in particular to a kit for detecting the virulence level of Klebsiella pneumoniae and a use method thereof, belonging to the technical field of biological detection. Background technique [0002] Klebsiella pneumoniae is a Gram-negative bacterium belonging to the family Enterobacteriaceae and the genus Klebsiella. It is widely distributed in nature. The upper respiratory tract and intestinal tract of the human body are conditional pathogenic bacteria, that is, under normal circumstances, Klebsiella pneumoniae does not cause disease, but it can cause infection when the body's immunity is low or long-term use of antibiotics leads to imbalance of flora. Klebsiella pneumoniae includes high-virulence Klebsiella pneumoniae and low-virulence (common) Klebsiella pneumoniae. The difference between the two is: 1. The former can infect young people without underlying ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/686C12Q1/10C12N15/11C12R1/22
CPCC12Q1/689C12Q1/686
Inventor 黄维邹全明张敏章金勇胡春霞惠真邓秋洋钟裕欣柳施一崔瑞勤
Owner SHENZHEN PEOPLES HOSPITAL
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