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Method for detecting toxicity of non-mucus klebsiella pneumoniae

A technology of Klebsiella pneumoniae and mucus, applied in the qualitative detection of non-mucinous Klebsiella pneumoniae virulence as high virulence or low virulence, and in the field of non-mucinous Klebsiella pneumoniae virulence detection, It can solve the problem that the virulence of non-mucinous Klebsiella pneumoniae cannot be identified, and achieve the effects of easy result judgment, small non-specific reaction and high specificity

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

AI Technical Summary

Problems solved by technology

In this technical solution, the amp R The gene is used as a biomarker, and specific primers are designed, and then based on the polymerase chain reaction, the qualitative detection of the virulence of Klebsiella pneumoniae of non-mucinous type is completed, so as to make up for the virulence detection method of Klebsiella pneumoniae in the prior art. It can be judged by whether it is hypermucinous type first, but the virulence of non-mucinous Klebsiella pneumoniae cannot be identified

Method used

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  • Method for detecting toxicity of non-mucus klebsiella pneumoniae
  • Method for detecting toxicity of non-mucus klebsiella pneumoniae
  • Method for detecting toxicity of non-mucus klebsiella pneumoniae

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Embodiment 1

[0042] non-mucinous Klebsiella pneumoniae amp R The application of the gene as a marker in the method for detecting the virulence of non-mucinous Klebsiella pneumoniae, if the marker is detected as positive, it is judged that the virulence of the non-mucinous Klebsiella pneumoniae is high virulence; otherwise, If the marker is tested negative, it is judged that the toxicity of non-mucinous Klebsiella pneumoniae is low toxicity;

[0043] said amp R The gene sequence is shown in SEQ ID NO.1.

Embodiment 2

[0045] A method for detecting the virulence of non-mucinous Klebsiella pneumoniae, comprising the steps of:

[0046] S1: Inoculate the non-mucinous Klebsiella pneumoniae to be detected in the culture medium, culture it, and take the culture solution with an OD600nm of 0.6 as the sample of the non-mucinous Klebsiella pneumoniae to be detected;

[0047] S2: Take at least four PCR tubes and number them respectively as positive control tubes, negative control tubes, experimental tubes and blank control tubes; prepare a PCR amplification system, among which,

[0048] Add 17 μL PCR reaction solution, 1 μL upstream primer, 1 μL downstream primer and 1 μL positive control to the positive control tube;

[0049] Add 17 μL PCR reaction solution, 1 μL upstream primer, 1 μL downstream primer and 1 μL negative control to the negative control tube;

[0050] Add 17 μL of PCR reaction solution, 1 μL of upstream primers, 1 μL of downstream primers and 1 μL of the non-mucinous Klebsiella pneumo...

Embodiment 3

[0057] On the basis of embodiment 2, the difference of this embodiment is:

[0058] Take the culture solution with OD600nm of 0.8 as the non-mucinous Klebsiella pneumoniae sample to be detected.

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Abstract

The invention discloses a method for detecting toxicity of non-mucus klebsiella pneumoniae, and belongs to the technical field of biological detection. The method comprises the following steps: takingan ampR gene sequence in a high-toxicity non-mucus klebsiella pneumoniae genome as a marker, designing a primer by using the specific sequence, and finally completing qualitative detection on the toxicity of the non-mucus klebsiella pneumoniae based on polymerase chain reaction. The method overcomes the defect that: the toxicity of the klebsiella pneumoniae can only be judged by judging whether the klebsiella pneumoniae is mucus or not and the toxicity difference in non-mucus strains cannot be distinguished in qualitative detection methods of the klebsiella pneumoniae in the prior art, so that the application values of the marker and the detection method for detecting the toxicity of the non-mucus klebsiella pneumoniae are guaranteed.

Description

technical field [0001] The invention relates to a method for detecting the virulence of non-mucinous Klebsiella pneumoniae, in particular to a qualitative detection method for detecting whether the virulence of non-mucinous Klebsiella pneumoniae is high or low, and belongs to biological detection technology field. 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 y...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/10C12R1/22
CPCC12Q1/689
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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