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Human sepsis pathogen detection kit and detection method

A detection kit and technology for sepsis, applied in biochemical equipment and methods, measurement/testing of microorganisms, resistance to vector-borne diseases, etc., can solve the problems of time-consuming, interference of blood culture positive rate and lack of diagnosis in sepsis Sensitivity and specificity and other issues, to achieve good sensitivity and specificity, high clinical application prospects, time and cost advantages

Active Publication Date: 2021-08-24
重庆市急救医疗中心重庆市第四人民医院重庆市急救医学研究所 +1
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AI Technical Summary

Problems solved by technology

At the same time, the positive rate of blood culture will be affected by factors such as blood collection volume, time of blood collection, frequency of blood collection, etc., which will interfere with the positive rate of blood culture in sepsis
In addition, it takes a long time to diagnose sepsis by blood culture, usually 48 to 72 hours
(2) Blood routine: It is currently considered clinically that white blood cells are 9 / L or greater than >20×10 9 / L should be highly suspected of infection, however, studies have shown that 50% of neonatal sepsis children's white blood cells can be within the normal range, and abnormal white blood cell counts can also appear in other pathological conditions, so the white blood cell count is very important for sepsis infection Diagnosis lacks sensitivity and specificity

Method used

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  • Human sepsis pathogen detection kit and detection method
  • Human sepsis pathogen detection kit and detection method
  • Human sepsis pathogen detection kit and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment one: the composition of kit

[0023] 1. Specific primer probe

[0024]

[0025]

[0026]

[0027] Note: where W is G or T: R is G or A; Y is C or T; M is A or C; S is G or C; D is A or G or T; K is G or T.

[0028] 2. The PCR reaction reagent is: TaqPath TM 1-Step Multiplex Master Mix (No ROX).

[0029] 3. Deionized water.

Embodiment 2

[0030] Embodiment two: DNA / RNA preparation

[0031] 1ml of human peripheral venous blood was collected according to medical routine, and EDTA was added for anticoagulation. Using Thermo's MagMAX TM CORE Nucleic Acid Purification Kit+MagMAX TM CORE Mastitis&PanbacteriaModule Extraction Kit, extract DNA and RNA.

Embodiment 3

[0032] Embodiment three: PCR reaction system preparation

[0033] Prepare the PCR reaction system according to the following system:

[0034] A total of 7 reaction systems (the total reaction system is 20 μL), which are respectively S1-S7 in the above table, are composed of mixed internal reference primers and probes. PCR reaction reagent: 10 μL; the concentration of each primer and probe is 10M, the volume of primers is 0.8 μL, and the volume of probe is 0.4 μL; sample template: 2 μL; deionized water is enough to make up 20 μL.

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Abstract

The invention provides a human sepsis pathogen detection kit and a corresponding detection method. The kit provided by the invention comprises primers and fluorescent probes fordetecting acinetobacter baumannii, klebsiella pneumoniae, streptococcus pneumoniae, bacteroides fragilis, staphylococcus aureus, pseudomonas aeruginosa, aeromonas hydrophila, escherichia coli, quail chicken enterococcus, enterobacter cloacae, stenotrophomonas maltophilia, coagulase negative staphylococcus, candida albicans, candida tropicalis, candida glabrata, cytomegalovirus, human herpesvirus type 4, human parvovirus B19, Candida parapsilosis, streptococcus.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection, in particular, the invention relates to a detection kit and a corresponding detection method for human sepsis pathogens. Background technique [0002] Sepsis is a syndrome in which infection causes dysregulation of the host response, leading to life-threatening organ dysfunction, and is a clinical syndrome with a high mortality rate. Sepsis not only seriously threatens human health, but also brings a huge economic burden to medical care. The fatality rate of sepsis in my country is higher than that in developed countries. A population-based epidemiological survey of sepsis found that 1,025,997 patients died of sepsis nationwide in 2015, accounting for 12.6% of the total case fatality rate. Another single-center retrospective study of 759 patients with severe sepsis from 2006 to 2016 showed that the mortality rate was as high as 36.6%. [0003] In May 2018, Chinese emergency...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6895C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/689C12Q1/6895C12Q1/701C12Q1/705C12Q1/686C12Q2563/107C12Q2545/114C12Q2537/143Y02A50/30
Inventor 李维邬文燕李逊姬晓雯庞博王芃芃
Owner 重庆市急救医疗中心重庆市第四人民医院重庆市急救医学研究所
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