Nucleic acid reagent, kit, system and method for detecting pathogenic bacteria of bloodstream infection

Abstract

The disclosure relates to a nucleic acid reagent for detecting bloodstream infection pathogenic bacteria. The nucleic acid reagent is characterized by comprising magnetic beads, which are separately stored independently or stored in a mutual-random-mixed manner, a capture probe shown in SEQ ID No. 1-6, a detection primer shown in SEQ ID No. 8-13 and a detection probe shown in SEQ ID No. 16-18, wherein the magnetic beads are modified with avidin, the capture probe is modified with biotin, and the avidin and the biotin can be specifically bonded. The nucleic acid reagent disclosed by the disclosure has high specificity and sensitivity to the bloodstream infection pathogenic bacteria and can achieve rapid detection on the bloodstream infection pathogenic bacteria.

Description

technical field [0001] The present disclosure relates to the field of biotechnology, in particular, to a nucleic acid reagent, kit and system for detecting pathogenic bacteria of bloodstream infection. Background technique [0002] Bloodstream infection (BSI) refers to a serious systemic infection caused by pathogenic microorganisms and toxins invading the blood circulation system. Pathogenic microorganisms that cause bloodstream infection mainly include bacteria, fungi, and viruses. In recent years, the global morbidity and mortality of BSI have been increasing year by year, and BSI progresses rapidly, which has become a common nosocomial infectious disease. Among the pathogens causing bloodstream infections, Gram-negative bacteria dominate, among which Acinetobacter baumannii (Ab), Pseudomonas aeruginosa (PA) and Klebsiella pneumoniae pneumoniae, KP) is the main pathogen causing bloodstream infection. [0003] At present, blood culture is the "gold standard" for diagnosi...

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Problems solved by technology

[0003] At present, blood culture is the "gold standard" for diagnosing BSI, but its detection cycle is long, usually 12-24 hours or even longer, and the sensitivity is poor. According to literature reports, the current positive rate of blood culture in China is only 16.9% , if the patient has previously used antibiotics, the positive rate will be further reduced, seriously affecting the patient's diagnosis and the best time for treatment

In addition, there are some non-culture auxiliary diagnosis, such as blood procalcitonin (PCT), neutrophil / lymphocyte ratio (neutrophil / lymphocyte ratio, NLR), C-reactive protein (C-reactive protein, CRP) and white blood cell count (white blood cell, WBC), etc., all have certain application value, but the conclusions are different and the specificity is poor

[0004] In addition, it has been reported that PCR technology can be used to directly detect the DNA of pathogenic bacteria in the blood of bloodstream infection. Therefore, direct detection of pathogenic bacteria from bloodstream infected blood has the following limitations: (1) pathogenic bacteria or their nucleic acid content is low, difficult to detect or inaccurate; (2) the current nucleic acid extraction technology steps are relatively Complicated, time-consuming, low yield, and unable to completely remove inhibitory factors affecting PCR in blood; (3) There are a large number of human genome backgrounds, which affect the detection of pathogenic bacteria or their nucleic acids; (4) Easy to be contaminated during PCR

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