Diagnostic method for early recognition of fetal intrauterine infection

Abstract

The invention provides a diagnostic method for early recognition of fetal intrauterine infection, comprising: collecting a blood sample of a pregnant woman and at least one fetal body fluid sample selected from an amniotic fluid sample or an umbilical cord blood sample; performing quantitative detection on a plurality of inflammatory factors in the blood sample and the fetal body fluid sample through a multichannel immunological detection technology, wherein the inflammatory factors comprise interleukin 6, C-reactive protein, tumor necrosis factor alpha, interleukin 1 beta and interleukin 8; various inflammatory factors in a pregnant woman blood sample and a fetal body fluid sample are comprehensively detected, simultaneous analysis of the multiple factors is achieved, abnormal fluctuation of the inflammatory factors is captured in real time in combination with dynamic monitoring of concentration changes, and clinical symptoms of the pregnant woman are analyzed through comparison with clinical symptoms of the pregnant woman. And diagnosis results can be recorded through an electronic health record system and early warning is generated, so that doctors are supported to judge and intervene in time, and the risks of missed diagnosis and misdiagnosis are effectively reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of medical diagnosis, and in particular to a diagnostic method for early identification of fetal intrauterine infection. Background Art

[0002] Fetal intrauterine infection refers to infection caused by pathogens transmitted from the mother to the fetus, which can cause fetal developmental abnormalities, premature birth, miscarriage, and even fetal death, seriously threatening the health and life safety of the fetus. Intrauterine infection is usually caused by bacteria, viruses or other pathogenic microorganisms. The early symptoms of infection are not obvious. Traditional diagnostic methods rely on the clinical manifestations of pregnant women or imaging examinations, but these methods have delays and limitations, and often cannot detect lesions in time in the early stages of intrauterine infection.

[0003] Traditional detection methods usually focus on a single biomarker or a specific pathogen. In practical applications, there are diagnostic limitations, which can easily lead to insufficient diagnostic sensitivity and accuracy. Conventional detection methods are highly dependent on single-collected samples and cannot dynamically capture the changing patterns of inflammatory factors over time, which can easily lead to a lag in early warning of intrauterine infection and delay the best time for intervention.

[0004] To this end, a diagnostic method for early identification of fetal intrauterine infection is proposed. Summary of the invention

[0005] In view of this, the present invention provides a diagnostic method for early identification of fetal intrauterine infection to solve or alleviate the technical problems existing in the prior art and at least provide a beneficial option;

[0006] The technical solution of the present invention is implemented as follows: a diagnostic method for early identification of fetal intrauterine infection, comprising:

[0007] collecting a blood sample and at least one fetal body fluid sample from the pregnant woman, wherein the fetal body fluid sample is selected from an amniotic fluid sample or an umbilical cord blood sample;

[0008] Quantitatively detecting multiple inflammatory factors in the blood sample and fetal body fluid sample by multi-channel immunological detection technology, wherein the inflammatory factors include interleukin 6, C-reactive protein, tumor necrosis factor α, interleukin 1β and interleukin 8;

[0009] During the detection process, inflammatory factors are measured in a layered and segmented manner, and high-concentration factors are quickly and quantitatively detected through a rapid screening module; low-concentration factors are then tested with higher sensitivity, and factors in different concentration ranges are tested in segments using immunoturbidimetry and enzyme-linked immunosorbent assay.

[0010] Based on the concentration changes of the inflammatory factors, the time series analysis method is used to analyze the trend of the concentration changes of the inflammatory factors over time. By calculating the concentration differences of each inflammatory factor at different time points, its change pattern is identified, and the concentration abnormality is evaluated in combination with the segmented detection results. If the concentration of a certain inflammatory factor increases continuously or rapidly in a short period of time, especially beyond the normal reference range, the abnormal changes related to fetal intrauterine infection are identified;

[0011] Further preferably, the multi-channel immunological detection technology adopts immunoassay technology to simultaneously perform quantitative detection of different inflammatory factors through multiple independent channels, including enzyme-linked immunosorbent assay ELISA, immunoturbidimetry and real-time fluorescence quantitative PCR, and simultaneously detect the concentrations of multiple inflammatory factors;

[0012] Further preferably, the concentration change of the inflammatory factor includes:

[0013] (1) By regularly testing inflammatory factors in maternal blood samples and fetal body fluid samples, the concentration differences of interleukin-6, C-reactive protein, tumor necrosis factor α, interleukin-1β, and interleukin-8 at different time points were calculated;

[0014] (2) Dynamic monitoring of the concentration of each inflammatory factor over time, using time series analysis to evaluate its changing trend in different stages of pregnancy, and identifying whether there are significant fluctuations or abnormal continuous increases. If a certain inflammatory factor concentration is found to increase continuously or accelerate within a certain period, it can be used as an early warning signal for fetal intrauterine infection;

[0015] (3) Based on the individualized normal physiological data of pregnant women and fetuses, a baseline model for changes in the concentration of inflammatory factors is established. If the concentration of one or more inflammatory factors exceeds the normal fluctuation range of the baseline model, it is considered an abnormal change.

[0016] Further preferably, based on the results of quantitative detection of inflammatory factor concentrations in maternal blood samples and fetal body fluid samples using multi-channel immunological detection technology, combined with the changing trend of the inflammatory factor concentrations, statistical methods are used to analyze the data to identify potential characteristics of fetal intrauterine infection.

[0017] Further preferably, the diagnostic method performs preliminary concentration screening on the maternal blood samples and fetal body fluid samples respectively before sample testing, and divides the samples into a high-concentration group and a low-concentration group according to the concentration range. The inflammatory factors in the high-concentration group are quickly screened by immunoturbidimetry, while the inflammatory factors in the low-concentration group are detected by the more sensitive enzyme-linked immunosorbent assay (ELISA).

[0018] Further preferably, after each inflammatory factor concentration test, the possibility of fetal intrauterine infection is evaluated by comparing it with the pregnant woman's body temperature, changes in vaginal secretions, and clinical symptoms of abnormal fetal movement. If abnormal changes in the inflammatory factor concentration are found, accompanied by worsening clinical symptoms of the pregnant woman, it is presumed that the fetus has suffered intrauterine infection.

[0019] Further preferably, the diagnostic method includes regular follow-up, and the data of each monitoring result and clinical symptoms are input into the electronic health record system to form a continuous data record. During the monitoring period, if obvious abnormal fluctuations in the concentration of inflammatory factors are detected, corresponding judgments are made in time through reports to provide early warning of the risk of fetal intrauterine infection.

[0020] Further preferably, the diagnostic method adds a target factor stabilizer during sample processing to protect the activity of the inflammatory factor, including:

[0021] (1) For target proteins that may be degraded in blood samples, anti-degradation enzyme reagents are added to inhibit protein hydrolysis;

[0022] (2) For oxidation-sensitive factors in fetal body fluid samples, antioxidants are added to prevent oxidation reactions;

[0023] (3) During transportation and storage, the samples are placed in a constant temperature environment and an appropriate amount of buffer is added to maintain pH stability.

[0024] The embodiment of the present invention has the following advantages due to the adoption of the above technical solution:

[0025] The present invention comprehensively detects multiple inflammatory factors in pregnant women's blood samples and fetal body fluid samples, realizes simultaneous analysis of multiple factors, and captures abnormal fluctuations of inflammatory factors in real time in combination with dynamic monitoring of concentration changes. It also improves the accuracy and sensitivity of diagnosis through comparative analysis with the clinical symptoms of pregnant women. The diagnostic results will be recorded and an early warning will be generated through the electronic health record system, supporting doctors to make timely judgments and interventions, solving the problems of single analysis and delayed diagnosis in traditional detection technologies, and effectively reducing the risks of missed diagnosis and misdiagnosis.

[0026] The present invention realizes rapid screening of high-concentration factors through a layered and segmented detection method, and accurately measures low-concentration factors through high-sensitivity detection technology, thereby reducing detection interference problems caused by excessive concentration spans and improving the reliability of detection results. At the same time, stabilizers are added during sample processing to ensure the activity of markers during the detection process and avoid detection errors caused by sample degradation or oxidation.

[0027] The above overview is for illustrative purposes only and is not intended to be limiting in any way; in addition to the illustrative aspects, embodiments, and features described above, further aspects, embodiments, and features of the present invention will be readily apparent by reference to the drawings and the following detailed description. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0029] Figure 1 is a diagnostic flow chart of the present invention; DETAILED DESCRIPTION

[0030] In the following, only certain exemplary embodiments are briefly described. As those skilled in the art will appreciate, the described embodiments may be modified in various different ways without departing from the spirit or scope of the present invention. Therefore, the drawings and description are to be regarded as illustrative in nature and not restrictive;

[0031] The embodiments of the present invention are described in detail below with reference to the accompanying drawings.

[0032] Example 1

[0033] like Figure 1 As shown, this embodiment discloses a diagnostic method for early identification of fetal intrauterine infection, comprising:

[0034] A blood sample and at least one fetal body fluid sample are collected from the pregnant woman, and the fetal body fluid sample is selected from an amniotic fluid sample or an umbilical cord blood sample; the collection of the pregnant woman's blood sample and the fetal body fluid sample is regularly monitored at different stages of pregnancy, and after each sample collection, multiple inflammatory factors in the sample are quantitatively tested, and the monitoring cycle is dynamically adjusted to adapt to the health status and clinical manifestations of the pregnant woman by analyzing the concentration change trend;

[0035] Through multi-channel immunological detection technology, multiple inflammatory factors such as interleukin-6, C-reactive protein, tumor necrosis factor α, interleukin-1β and interleukin-8 in maternal blood samples and fetal body fluid samples are quantitatively detected. The detection method is based on a multi-channel detection platform and uses immunological analysis to achieve simultaneous detection of multiple inflammatory factors in a single system, thereby meeting the high clinical requirements for detection efficiency, accuracy and sensitivity. The characteristic of multi-channel immunological detection is that it uses multiple independent detection channels to specifically detect different inflammatory factors. Each channel uses strictly calibrated antibodies or nucleic acid probes to accurately bind and identify the target factors.

[0036] Specifically, ELISA is used to quantitatively analyze the inflammatory factors in the sample. ELISA achieves the capture of the target factor in the microplate through the specific reaction of antigen-antibody, and detects and quantifies it through enzymatic reaction color or fluorescence signal. Different antibodies can be pre-coated on the ELISA microplate, and each well targets different inflammatory factors to achieve parallel detection of multiple targets. Secondly, the immunoturbidimetric method is an optical detection based on the formation of immune complexes after antigen-antibody reaction, which is particularly suitable for rapid screening of target factor concentrations. By detecting changes in light scattering intensity, the concentration of inflammatory factors can be quickly quantified, which is particularly suitable for samples with large detection volumes. Real-time fluorescence quantitative PCR can further verify abnormal changes in protein levels by amplifying and detecting specific nucleic acid sequences of inflammatory factors.

[0037] During the detection process, the inflammatory factors were accurately measured in a stratified and segmented manner to solve the problem of detection interference caused by the large difference in the concentration range of the factors. The high-concentration inflammatory factors in the sample were preliminarily detected through the rapid screening module. The rapid screening module used immunoturbidimetry to quantitatively measure the high-concentration factors. Based on the influence of the immune complex formed after the combination of antigen and antibody on the light scattering signal, it is suitable for the rapid analysis of high-concentration markers and can provide accurate concentration data in a short time. Then, for the low-concentration inflammatory factors in the sample, the more sensitive enzyme-linked immunosorbent assay (ELISA) was used for detection. The ELISA method combines antigens with specific antibodies and then amplifies the signal through enzymatic reactions to achieve accurate detection of low-concentration factors. To ensure the accuracy of stratified and segmented detection, the samples are divided into high-concentration group and low-concentration group according to the concentration range before entering the detection channel. The high-concentration group directly enters the rapid screening module, while the low-concentration group needs to be appropriately diluted before entering the ELISA detection system. Through the stratified and segmented measurement strategy, the detection of high-concentration factors avoids the problem of signal saturation, and the detection of low-concentration factors also avoids the decrease in sensitivity caused by the interference of high-concentration factors.

[0038] In the detection process, the sample is pretreated to remove lipid or protein degradation products in the plasma sample, and the sample is injected into the multi-channel detection system. The reaction modules in each channel complete antibody binding, signal amplification and detection in turn. The final detection data is automatically read by the system and integrated on the data analysis platform to form a multi-dimensional detection result. Combined with the concentration changes of inflammatory factors and their dynamic performance during pregnancy, the early signals of fetal intrauterine infection can be effectively identified. This method is used for the joint detection of maternal blood samples and fetal body fluid samples. It can not only detect the concentration changes of a certain inflammatory factor alone, but also conduct a joint analysis of multiple factors.

[0039] Changes in the concentrations of inflammatory factors include:

[0040] (1) By regularly testing inflammatory factors in maternal blood samples and fetal body fluid samples, the concentration differences of interleukin-6, C-reactive protein, tumor necrosis factor α, interleukin-1β, and interleukin-8 at different time points were calculated;

[0041] (2) Dynamically monitor the concentration changes of each inflammatory factor and use time series analysis to evaluate its changing trend at different stages of pregnancy. Time series analysis combined with multi-point sampling data can identify long-term patterns and short-term fluctuations in concentration changes;

[0042] For example, if the concentration of an inflammatory factor continues to rise or reaches a peak in a short period of time, it indicates that there may be an acute inflammatory response; if the concentration level fluctuates periodically, its pathological significance needs to be judged in combination with other clinical indicators of the pregnant woman. If the concentration of an inflammatory factor is detected to increase continuously or accelerate in a specific cycle, especially if it exceeds the normal reference range, it is considered a high-risk signal for intrauterine infection.

[0043] (3) Based on the collected inflammatory factor concentration data, combined with the individualized normal physiological parameters of pregnant women and fetuses, a baseline model of concentration changes is established, which is constructed from the previous health records of pregnant women and fetuses and reference data of similar groups. It is used to define the fluctuation range and change trend of each inflammatory factor under normal physiological conditions. By comparing the test data with the baseline model, abnormal fluctuations in concentration can be quickly identified;

[0044] Based on the results of quantitative detection of inflammatory factor concentrations in maternal blood samples and fetal body fluid samples using multi-channel immunological detection technology, combined with the changing trend of inflammatory factor concentrations, statistical methods are used to conduct a comprehensive analysis of the data to identify potential characteristics of fetal intrauterine infection; while analyzing, the test results are compared with clinical symptoms such as the pregnant woman's body temperature, changes in vaginal secretions, and abnormal fetal movements. If the concentration of inflammatory factors changes abnormally and is accompanied by aggravated clinical symptoms of the pregnant woman, it is inferred that the fetus has had intrauterine infection;

[0045] By comparing the concentration values ​​at adjacent detection time points, the difference value △C=Ct2-Ct1 is calculated, where Ct2 is the current concentration and Ct1 is the concentration at the previous time point. The concentration change trend is modeled based on time series analysis. During the analysis, the normal range of inflammatory factors is obtained by large-scale data statistics.

[0046] For example, the normal reference range of CRP is 0.1-10mg / L, and that of IL-6 is 0-7pg / mL. If the calculated △C continues to be positive and the value increases, or if it is found that the concentration of a factor reaches more than twice its reference range in a short period of time, such as CRP>20mg / L or IL-6>15pg / mL, it is judged to be significantly abnormal. At the same time, combined with the sensitivity data verification of the segmented detection results, it ensures that abnormal changes in high-concentration and low-concentration factors are accurately captured.

[0047] Finally, each monitoring result and the clinical symptom data of the pregnant woman are input into the electronic health record system to form a continuous data record. If an obvious abnormal fluctuation in the concentration of inflammatory factors is detected during the monitoring period, the system will automatically generate a report to remind the doctor to make a timely judgment and warn the possibility of fetal intrauterine infection in advance. The health record generated during the diagnosis process can be used as a reference for the doctor's follow-up and subsequent diagnosis;

[0048] After each test of the inflammatory factor concentration, a comprehensive assessment of the pregnant woman's health status and the fetal intrauterine environment is conducted based on the pregnant woman's stage of pregnancy and changes in the test results, combined with the doctor's diagnostic experience.

[0049] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art can easily think of various changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A diagnostic method for early identification of fetal intrauterine infection, characterized in that: include: collecting a blood sample and at least one fetal body fluid sample from the pregnant woman, wherein the fetal body fluid sample is selected from an amniotic fluid sample or an umbilical cord blood sample; Quantitatively detecting multiple inflammatory factors in the blood sample and fetal body fluid sample by multi-channel immunological detection technology, wherein the inflammatory factors include interleukin 6, C-reactive protein, tumor necrosis factor α, interleukin 1β and interleukin 8; During the detection process, inflammatory factors are measured in a layered and segmented manner, and high-concentration factors are quickly and quantitatively detected through a rapid screening module; low-concentration factors are then tested with higher sensitivity, and factors in different concentration ranges are tested in segments using immunoturbidimetry and enzyme-linked immunosorbent assay. Based on the changes in the concentration of the inflammatory factors, the time series analysis method is used to analyze the trend of the changes in the concentration of the inflammatory factors over time. By calculating the concentration differences of each inflammatory factor at different time points, its change pattern is identified, and the concentration abnormality is evaluated in combination with the segmented detection results. If the concentration of a certain inflammatory factor increases continuously or rapidly in a short period of time, especially if it exceeds the normal reference range, abnormal changes related to fetal intrauterine infection are identified.

2. A diagnostic method for early identification of fetal intrauterine infection according to claim 1, characterized in that: The multi-channel immunological detection technology adopts immunoassay technology to simultaneously perform quantitative detection of different inflammatory factors through multiple independent channels, including enzyme-linked immunosorbent assay (ELISA), immunoturbidimetry and real-time fluorescence quantitative PCR, and simultaneously detects the concentrations of multiple inflammatory factors.

3. A diagnostic method for early identification of fetal intrauterine infection according to claim 1, characterized in that: The changes in the concentration of the inflammatory factors include: (1) By regularly testing inflammatory factors in maternal blood samples and fetal body fluid samples, the concentration differences of interleukin-6, C-reactive protein, tumor necrosis factor α, interleukin-1β, and interleukin-8 at different time points were calculated; (2) Dynamic monitoring of the concentration of each inflammatory factor over time, using time series analysis to evaluate its changing trend in different stages of pregnancy, and identifying whether there are significant fluctuations or abnormal continuous increases. If a certain inflammatory factor concentration is found to increase continuously or accelerate within a certain period, it can be used as an early warning signal for fetal intrauterine infection; (3) Based on the individualized normal physiological data of pregnant women and fetuses, a baseline model for changes in the concentration of inflammatory factors is established. If the concentration of one or more inflammatory factors exceeds the normal fluctuation range of the baseline model, it is considered an abnormal change.

4. A diagnostic method for early identification of fetal intrauterine infection according to claim 1, characterized in that: Based on the results of quantitative detection of inflammatory factor concentrations in maternal blood samples and fetal body fluid samples using multi-channel immunological detection technology, combined with the changing trend of the inflammatory factor concentrations, statistical methods were used to analyze the data to identify potential characteristics of fetal intrauterine infection.

5. A diagnostic method for early identification of fetal intrauterine infection according to claim 4, characterized in that: Before sample testing, the diagnostic method conducts preliminary concentration screening on pregnant woman's blood samples and fetal body fluid samples, and divides the samples into high-concentration group and low-concentration group according to the concentration range. The inflammatory factors in the high-concentration group are quickly screened by immunoturbidimetry, while the inflammatory factors in the low-concentration group are detected by the more sensitive enzyme-linked immunosorbent assay (ELISA).

6. A diagnostic method for early identification of fetal intrauterine infection according to claim 1, characterized in that: After each inflammatory factor concentration test, the possibility of fetal intrauterine infection is assessed by comparing it with the pregnant woman's body temperature, changes in vaginal secretions, and clinical symptoms of abnormal fetal movement. If abnormal changes in the inflammatory factor concentration are found and accompanied by worsening clinical symptoms of the pregnant woman, it is presumed that the fetus has had intrauterine infection.

7. A diagnostic method for early identification of fetal intrauterine infection according to claim 1, characterized in that: The diagnostic method includes regular follow-up, and inputting the data of each monitoring result and clinical symptoms into the electronic health record system to form a continuous data record. During the monitoring period, if obvious abnormal fluctuations in the concentration of inflammatory factors are detected, corresponding judgments are made in time through reports to provide early warning of the risk of fetal intrauterine infection.

8. A diagnostic method for early identification of fetal intrauterine infection according to claim 1, characterized in that: The diagnostic method adds a target factor stabilizer during sample processing to protect the activity of inflammatory factors, including: (1) For target proteins that may be degraded in blood samples, anti-degradation enzyme reagents are added to inhibit protein hydrolysis; (2) For oxidation-sensitive factors in fetal body fluid samples, antioxidants are added to prevent oxidation reactions; (3) During transportation and storage, the samples are placed in a constant temperature environment and an appropriate amount of buffer is added to maintain pH stability.

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