Application of Abeta1-42 and CD64 combination in preparation of reagent for early detection of sepsis

Abstract

The invention provides application of a combination of A beta1-42 and CD64 in preparation of a reagent for early detection of sepsis, and belongs to the technical field of biology, the diagnosis value of combined detection of A beta1-42 and CD64 is significantly higher than that of a single index, a synergistic effect is generated, and the technical problem that a single marker is insufficient in sensitivity or specificity is solved. The CD64 is quick in response, the Abeta1-42 is closely associated with the pathological process, and the combination is beneficial to identifying high-risk patients in the early stage of diseases and striving for precious intervention time for clinic. The marker combination not only can be used for qualitative diagnosis, but also can be used for risk stratification and prognosis judgment, and has comprehensive functions, and the quantification level of the marker combination is positively correlated with the severity of diseases.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, and in particular relates to the application of Aβ1-42 and CD64 in the preparation of reagents for early detection of sepsis. Background Technology

[0002] Sepsis is a systemic inflammatory response syndrome caused by infection, which can further develop into severe sepsis, septic shock, and multiple organ dysfunction syndrome. The pathological mechanism of this disease is complex, with the core issue being the uncontrolled response of the body to infection, leading to widespread activation of the immune and coagulation systems, endothelial damage, and organ dysfunction. Patients clinically present with fever or hypothermia, tachycardia, tachypnea, and altered consciousness. The condition often progressively worsens, rapidly leading to life-threatening complications such as acute kidney injury, acute respiratory distress syndrome, and circulatory failure. Due to its insidious onset, rapid progression, and high mortality rate, sepsis has become one of the leading causes of death in intensive care units.

[0003] Early diagnosis and intervention of sepsis are crucial for improving prognosis, but clinical practice still faces significant challenges. Currently, there are no biomarkers directly applicable to the early diagnosis of sepsis. PCT and CRP are two of the most commonly used biomarkers for assessing infection in clinical practice, but their levels are generally only used to determine the strength of evidence of infection, not to directly diagnose sepsis. In recent years, many new biomarkers have been reported in research papers, but most cannot yet be directly applied clinically. Furthermore, PCT and CRP have limitations in early sensitivity and specificity: PCT may not rise significantly in non-bacterial infections, localized infections, or inflammatory states; CRP has a relatively delayed response and is affected by various non-infectious factors. In addition, sepsis is highly heterogeneous; different pathogens, infection sites, and host immune status lead to complex and variable biomarker expression profiles, making it difficult for a single indicator to achieve accurate early identification, risk stratification, and prognostic assessment.

[0004] Therefore, there is an urgent need in this field to discover novel, efficient, and especially synergistic diagnostic biomarkers or combinations that can reflect specific pathophysiological processes of sepsis, in order to overcome the bottlenecks of existing diagnostic technologies in terms of early detection and accuracy, and provide a more reliable basis for timely clinical intervention and individualized management. Summary of the Invention

[0005] In view of this, the object of the present invention is to provide the application of Aβ1-42 and CD64 in combination in the preparation of a reagent for the early detection of sepsis.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides the application of Aβ1-42 and CD64 in the preparation of reagents for the early detection of sepsis.

[0007] This invention provides a kit for early detection of sepsis, the kit comprising reagents for detecting Aβ1-42 and CD64.

[0008] Preferably, when the CD64 index is >3.30 and Aβ1-42 is >50 pg / mL, it indicates sepsis.

[0009] Preferably, the sample for detecting sepsis includes whole blood, serum, or plasma.

[0010] Preferably, the method for detecting Aβ1-42 and CD64 includes flow cytometry or immunoassay.

[0011] Compared with the prior art, the present invention has the following beneficial effects: This invention is the first to combine CD64 and Aβ1-42 for the diagnosis of sepsis. By creatively combining CD64 and Aβ1-42, two biomarkers that play different roles and have different dynamic characteristics in the pathophysiology of sepsis, ultra-early and highly accurate early warning of sepsis is achieved, effectively solving the core bottleneck of existing diagnostic technologies where sensitivity and specificity cannot be simultaneously achieved during the disease window period. Experiments have shown that the diagnostic value (AUC) of the combined detection of the two is significantly higher than that of a single indicator, producing a synergistic effect and solving the technical problem of insufficient sensitivity or specificity of a single biomarker. CD64 responds rapidly, and Aβ1-42 is closely related to the pathological process. This combination helps to identify high-risk patients in the early stage of the disease, buying valuable time for clinical intervention. This biomarker combination can not only be used for qualitative diagnosis, but its quantitative level is also positively correlated with the severity of the disease, making it suitable for risk stratification and prognosis assessment, demonstrating comprehensive functionality. Attached Figure Description

[0012] Figure 1 The ROC results for CD64 are shown in the figure. Figure 2 The ROC results for Aβ1-42 are shown in the figure. Figure 3 The ROC results for the combined detection of CD64 and Aβ1-42 are shown in the figure. Detailed Implementation

[0013] This invention provides the application of Aβ1-42 and CD64 in the preparation of reagents for the early detection of sepsis.

[0014] This invention provides a kit for early detection of sepsis, the kit comprising reagents for detecting Aβ1-42 and CD64.

[0015] In this invention, the samples used to detect sepsis include whole blood, serum, or plasma.

[0016] In this invention, it is preferred that a CD64 index > 3.30 and an Aβ1-42 > 50 pg / mL indicate sepsis.

[0017] In this invention, the method for detecting Aβ1-42 and CD64 includes flow cytometry or immunoassay.

[0018] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0019] Example 1

[0020] Four hundred infected patients from the ICUs of West China Hospital of Sichuan University (including the main campus, Tianfu campus, and Shangjin campus) were included in the study, along with 150 healthy individuals selected through health checkups at the West China Medical Examination Center of Sichuan University as controls. All participants signed informed consent forms before specimen collection. Disease progression was tracked, and the Sepsis-3 criteria were used to diagnose whether patients progressed to sepsis. Patients diagnosed with sepsis were followed up at 28 and 96 days to track their outcomes. Using the single-molecule droplet digital PCR system developed by Hangzhou Leading Gene Technology Co., Ltd., single-cell sequencing was performed, followed by whole-genome transcriptome sequencing of peripheral blood, spleen, lymph nodes, and bone marrow. Two biomarkers, CD64 and Aβ1-42, were identified. CD64, a rapidly reacting protein on the surface of neutrophils, was significantly upregulated within 4-6 hours under bacterial infection stimulation. An elevated level of these indicators signifies a strong immune response initiated by the innate immune system, a signal that precedes clinically apparent deterioration of organ function. Furthermore, the concentration of Aβ1-42, a molecule closely associated with neuroinflammation and endothelial damage, rises abnormally in the early stages of infection due to impaired clearance or increased production, indicating that the infection has triggered a systemic pathological disorder that may lead to organ damage. When both indicators simultaneously exceed specific thresholds (CD64 index > 3.30, Aβ1-42 > 50 pg / mL), it suggests that the patient is not only in a state of strong immune activation but has also entered a high-risk pathological stage prone to organ dysfunction. By detecting this combined signal, it is possible to identify high-risk individuals most likely to progress to sepsis during the initial assessment of suspected infected patients, before the traditional diagnostic criteria for sepsis are fully met, thus enabling early diagnosis and intervention.

[0021] Example 2

[0022] To simplify the detection method and verify the effectiveness of the biomarkers screened in Example 1, 216 new patients with sepsis were re-screened from the ICU of West China Hospital of Sichuan University, and 50 new healthy individuals screened from health checkups at West China Health Examination Center of Sichuan University served as controls. All participants signed informed consent forms before specimen collection. Disease progression was tracked, and the Sepsis-3 criteria were used to diagnose whether patients had progressed to sepsis. Patients diagnosed with sepsis were followed up for 28 and 96 days to track their outcomes. Flow cytometry was used to determine the concentrations of CD64 index and Aβ1-42 in peripheral blood (CD64 index > 3.30, Aβ1-42 > 50 pg / mL). The effectiveness of individual and combined detection of CD64 and Aβ1-42 was analyzed. The AUC, sensitivity, and specificity of individual and combined detection of CD64 and Aβ1-42 are detailed in Table 1. Figures 1-3 .

[0023] Table 1. AUC, sensitivity, and specificity of CD64 and Aβ1-42 alone and in combination.

[0024] Note: AUC is the area under the receiver operating curve (ROC). The closer the AUC is to 1, the better or more accurate the product's diagnostic performance.

[0025] The results showed that the diagnostic value (AUC) of the combined detection of the two biomarkers was significantly higher than that of the single biomarker, producing a synergistic effect.

[0026] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Application of Aβ1-42 and CD64 in the preparation of reagents for early detection of sepsis.

2. A kit for early detection of sepsis, characterized in that, The kit includes reagents for detecting Aβ1-42 and CD64.

3. The reagent kit according to claim 2, characterized in that, When the CD64 index is >3.30 and Aβ1-42 is >50 pg / mL, it indicates sepsis.

4. The reagent kit according to claim 3, characterized in that, The samples used to detect sepsis include whole blood, serum, or plasma.

5. The reagent kit according to claim 2, characterized in that, The methods for detecting Aβ1-42 and CD64 include flow cytometry or immunoassay.

Images