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Detection system for urinary system infection pathogens and kit and application thereof

A urinary system infection and pathogen detection technology, applied in the biological field, can solve the problems of high drug resistance rate of strains, incomplete coverage, and increased medical burden on patients, so as to ensure specificity and sensitivity, ensure accuracy and reliability, and reduce medical costs. cost effect

Pending Publication Date: 2021-05-28
HUADONG HOSPITAL +1
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many deficiencies in culture identification: ①Long time-consuming: ordinary bacterial culture incubation takes 16 to 24 hours, and it takes even longer for pathogens with slow growth and complex nutritional requirements; biochemical identification and drug sensitivity test are required after the colony grows, It takes at least 3 working days; ②Incomplete coverage and low positive detection rate: Because ordinary culture is mostly aimed at a single pathogen, and UTI also has difficult-to-cultivate but extremely important pathogenic bacteria such as Mycobacterium tuberculosis and Mycoplasma, which often lead to false negative
However, the current conventional detection methods have the disadvantages of low detection rate, long time-consuming, especially the inability to accurately identify multiple pathogens at the same time, and cannot provide timely, comprehensive and accurate pathogen diagnosis basis for clinical practice, leading to the widespread use of broad-spectrum Empirical treatment with antibacterial drugs leads to low curative effect, failure to control the progress of urinary tract infection in time, high drug resistance rate of induced strains, and increased medical burden on patients, etc.

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  • Detection system for urinary system infection pathogens and kit and application thereof
  • Detection system for urinary system infection pathogens and kit and application thereof
  • Detection system for urinary system infection pathogens and kit and application thereof

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Embodiment 1

[0059] 1. The composition of the kit

[0060] The urinary system infection pathogen detection kit of this embodiment includes: multiplex PCR master mix, multiplex PCR enzyme solution, primer mixture, positive control, negative control, system quality control internal reference (IC) and nuclease-free pure water. The multiplex PCR master mix is ​​prepared by mixing 10×PCR buffer, MgCl2 and dNTPs. The multiplex PCR enzyme solution is a mixture of hot start DNA polymerase and UNG enzyme.

[0061] Both the multiplex PCR master mix and the multiplex PCR enzyme solution were from Roche Company (Catalog No.: 210212).

[0062] The positive control is a plasmid mix that includes all gene targets of interest.

[0063] The negative control was nuclease-free ultrapure water.

[0064]The primer mixture includes primer pairs for detecting 18 kinds of pathogen target genes, primer pairs for detecting human DNA internal reference and primer pair for detection system quality control internal...

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Abstract

The invention relates to a urinary system infection pathogen detection system and a kit and application thereof, the detection system comprises 20 pairs of primers, including 18 pairs of detection primers, one pair of human DNA internal reference primers and one pair of system quality control internal reference primers. According to the detection system for the drug-resistant gene of the urinary system infection pathogen as well as the kit and the application of the detection system, conventional detection is not needed, and synchronous identification and quantitative analysis of various pathogens can be directly carried out on a urine sample in the same reaction system. The system has the advantages of strong specificity, high sensitivity, good repeatability, rapidness and accuracy, provides comprehensive, accurate and low-cost etiological diagnosis for clinic in the first time, and provides important reference for individualized medication and precise medical treatment.

Description

technical field [0001] The invention relates to a multiple gene detection product and a detection system used in the product, belonging to the field of biotechnology. Background technique [0002] Urinary system infection, also known as urinary tract infection (Urinary Tract Infections, UTI), is one of the most common infectious diseases clinically. A variety of pathogens can invade the urinary system and grow and reproduce in large numbers, causing clinical symptoms such as frequent urination, urgency, dysuria, dysuria, and waist and leg pain. In severe cases, severe diseases such as sepsis and acute and chronic renal dysfunction may develop; However, urogenous sepsis and septic shock progress very rapidly, and the fatality rate is as high as 28.3% to 41.1%. UTI accounts for about 40% of nosocomial infections in foreign countries, ranking first; while the incidence of UTI in my country is as high as 20.8% to 31.7%, second only to respiratory tract infections, and more than...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/6895C12Q1/6848
CPCC12Q1/689C12Q1/6895C12Q1/686Y02A50/30
Inventor 张艳梅赵虎孙召洋吴勇南丽余丁
Owner HUADONG HOSPITAL
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