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A molecular detection kit for hepatic abscess-causing Klebsiella pneumoniae and its application

A technology of Klebsiella pneumoniae and detection kit, which is applied in the field of disease diagnosis to achieve the effect of improving the success rate of treatment

Active Publication Date: 2019-05-31
NANJING FIRST HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is currently no quick and easy method for the detection and identification of Klebsiella pneumoniae causing liver abscess

Method used

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  • A molecular detection kit for hepatic abscess-causing Klebsiella pneumoniae and its application
  • A molecular detection kit for hepatic abscess-causing Klebsiella pneumoniae and its application
  • A molecular detection kit for hepatic abscess-causing Klebsiella pneumoniae and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Development and validation of specific sequences:

[0034]The screening process of specific gene sequences combines the next-generation sequencing technology (Next-Generation Sequencing, NGS), comparative genomics (Comparative Genomics) and bioinformatics (Bioinformatics) methods, and uses scientific statistical methods to analyze sequences in multiple The specificity of the distribution in the Klebsiella pneumoniae genome was tested, and PCR validation was performed in the isolated Klebsiella pneumoniae and non-hepatoabscess Klebsiella pneumoniae to determine the sequence in Klebsiella pneumoniae. Specificity in Lebsiella. The specific implementation process is as follows:

[0035] Two strains of Klebsiella pneumoniae and one strain of Klebsiella pneumoniae isolated from mainland China were selected for whole-genome sequencing, and the sequencing results were spliced ​​by SPAdes (v3. Analysis platform for gene prediction and functional annotation. The obtained seque...

Embodiment 2

[0043] Gene-specific verification:

[0044] The specific steps for the verification of pagO and luxR-induced liver abscess Klebsiella pneumoniae-specific genes in clinical isolates are as follows:

[0045] 1. Select clinically isolated hepatic abscess and non-hepatic abscess Klebsiella pneumoniae bacterial strains for detection of pagO (shown in SEQ ID N O.1) and luxR (shown in SEQ ID NO.2) in causing hepatic abscess pneumonia Specificity in Klebsiella. 40 strains of Klebsiella pneumoniae isolated from the abscess of patients with liver abscess were selected as the detection group (hepatic abscess group), and 36 strains of Klebsiella pneumoniae isolated from non-Klebsiella pneumoniae patients with liver abscess were selected as negative controls group (non-hepatic abscess-induced group). At the same time, other common pathogenic bacteria (other pathogenic bacteria groups) isolated clinically were also selected to further verify the specificity of these factors in Klebsiella ...

Embodiment 3

[0061] Application of pagO and luxR sequences or primers designed based on the sequences in the preparation of Klebsiella pneumoniae liver abscess diagnostic kit:

[0062] The design idea of ​​this experiment is as follows: the clinically obtained blood culture samples are divided into a blood culture positive group and a healthy person negative group, where the blood culture positive group refers to all samples with bacteria cultured in the blood, not limited to the blood cultured with pneumonia. Lebsiella, including cultures of other pathogenic bacteria. Use PCR to detect the presence of specific factors (i.e. pagO and luxR) in the positive blood culture sample and the negative blood culture sample, and finally analyze the corresponding analysis of the PCR result and the patient's final clinical diagnosis, so as to judge that the method provided by the present invention is effective against pneumonia. Detection rate of Lebsiella liver abscess.

[0063] 1. Collection of clin...

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PUM

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Abstract

The invention discloses a molecular detection kit for klebsiella pneumoniae causing hepatic abscess and an application thereof. The invention provides gene specific sequences pagO and luxR of klebsiella pneumoniae causing hepatic abscess, two genes are specially existed in the klebsiella pneumoniae causing hepatic abscess, and can be taken as a diagnostic marker of klebsiella pneumoniae causing hepatic abscess. The primers of design of two genes can be used for preparing the molecular detection kit for klebsiella pneumoniae causing hepatic abscess, the kit can be used for diagnosis or early diagnosis of the molecular detection kit for klebsiella pneumoniae causing hepatic abscess, and has great commercialized exploitation value.

Description

technical field [0001] The invention relates to the field of disease diagnosis, in particular to a hepatic abscess-causing Klebsiella pneumoniae molecular detection kit and application thereof. Background technique [0002] Klebsiella pneumoniae (K.pneumoniae subsp.pneumonia) belongs to Enterobacteriaceae bacteria. It often parasitizes human skin, nasopharynx and intestinal tract, and is an opportunistic pathogen. Clinically, it can cause pneumonia and urinary tract infection, etc. ). From 1980 to 1990, Liu et al. in Taiwan reported 7 cases of severe liver abscess with endophthalmitis caused by highly virulent Klebsiella pneumoniae for the first time (Liu Y, Cheng D, Lin C. Klebsiellapneumoniae liver abscess associated with septic endophthalmitis. Arch InternMed 1986;146:1913–16). Since then, Klebsiella pneumoniae liver abscesses have become a global epidemic, especially in Asia. Compared with common Klebsiella pneumoniae, Klebsiella pneumoniae causing liver abscess has...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/31C12N15/11C12Q1/689C12Q1/04
CPCC12Q1/689
Inventor 张彦亮陶臻黄岩俞杨黄金伟江建平叶美萍
Owner NANJING FIRST HOSPITAL
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