CN112442543A

CN112442543A - Method for detecting stability of arterial plaque

Info

Publication number: CN112442543A
Application number: CN201910794409.8A
Authority: CN
Inventors: 孙余华
Original assignee: Individual
Current assignee: Individual
Priority date: 2019-08-27
Filing date: 2019-08-27
Publication date: 2021-03-05

Abstract

The invention relates to the field of testing medicine and cardiovascular and cerebrovascular diseases. The invention relates to a method for testing human peripheral blood, in particular to a method for testing the stability of arterial plaque, which is used for testing the chlamydia pneumoniae genome-specific nucleic acid sequence in the peripheral blood to test the chlamydia pneumoniae infection in the peripheral blood and in the arterial plaque so as to judge the stability of the arterial plaque. The following three aspects are specified: chlamydia pneumoniae infection in peripheral blood as a new blood risk factor for development of arterial plaque development and associated disease pathogenesis; the novel method for detecting the stability of the arterial plaque also classifies and stages the inflammatory response of the arterial plaque according to the infection condition and the infection degree of chlamydia pneumoniae in the arterial plaque of an individual; and determining the application method of the medicine program for stabilizing the arterial plaque and reducing the inflammatory response according to the detected human peripheral blood and the chlamydia pneumoniae infection condition in the arterial plaque. The method has good social value and economic value.

Description

Method for detecting stability of arterial plaque

Technical Field

The invention relates to a method for testing extracted human peripheral blood, in particular to a method for detecting chlamydia pneumoniae infection in arterial plaque by testing chlamydia pneumoniae in human peripheral blood through a PCR (polymerase chain reaction) technology, and further relates to a method for detecting arterial plaque stability and determining application in implementing a plaque stabilizing drug scheme. The invention relates to the field of testing medicine and cardiovascular and cerebrovascular diseases.

Background

Arterial plaque, also known as atherosclerotic plaque, is a major cause of cardiovascular and cerebrovascular diseases. Currently, the blood risk factor detection indexes related to the development of arterial plaque include high total cholesterol, high low density lipoprotein cholesterol, high small and dense low density lipoprotein cholesterol, high triglyceride, hyperglycemia, homocysteine, high lipoprotein (a) or hypersensitive C-reactive protein level, etc. (Yuhua Sun, et al. Clin. Cardiol, 2007, 30: 576-580). These blood risk factors are mostly normal components of human blood, but their values are higher than normal levels, and nearly half of arterial plaque and clinical cardiovascular and cerebrovascular disease patients have the above blood indexes in the normal range. In fact, low or high levels of any normal components in human blood are not beneficial for cardiovascular and cerebrovascular diseases (Yuhua Sun, Yuejin Yang. future Lipidol, 2008, 3: 127-. Therefore, there are other blood risk factors associated with the development of arterial plaque development and the development of cardiovascular and cerebrovascular diseases.

Most arterial plaque lesions are characterized locally by intimal thickening, plaque formation, and luminal narrowing. The occurrence of clinical diseases related to arterial plaque, particularly cardiovascular and cerebrovascular diseases, is influenced by factors such as arterial plaque load, lumen stenosis degree and the like, and is directly related to rupture or erosion of unstable arterial plaque and thrombosis. At present, the examination of whether the artery plaque exists, calcified and non-calcified lesion exists and the degree of artery lumen stenosis is mainly carried out by the techniques of carotid artery ultrasound, coronary artery CT, coronary artery angiography or cerebrovascular angiography and the like. Whether the arterial plaque is a thin fiber cap plaque, whether the rupture or the healing occurs, whether the plaque has vulnerable characteristics such as a new blood vessel and the like, the arterial plaque is mainly examined by technologies such as intravascular ultrasound, optical coherence tomography, nuclear magnetic resonance and the like, the morphological structure of the plaque can be directly observed through the examination, and the stability of the arterial plaque can be judged. However, we need to improve the understanding of the inflammatory nature of arterial plaque from the viewpoint of theoretical innovation and provide a more prospective and effective method for recognizing the stability of arterial plaque.

The pathological nature of arterial plaque is inflammatory reaction (Sun Yuhua, Wang Feng Lian. division of foreign medical immunology 1992, 1: 16-19), which occurs and progresses through vascular endothelial injury, local deposition of blood substances, proliferation and secretory transformation of vascular smooth muscle cells, various inflammatory factors produced by inflammatory cells, inflammatory exudation, cellular necrosis, fibrosis and calcification. The existence of microorganisms is also proved in the local part of the lesion of the artery plaque. Most scholars believe that the microorganisms are just "innocent bystanders" and that arterial plaque is a sterile inflammation. The inventors believe that "foreign, non-human" microorganisms within the arterial plaque are involved in the development of inflammation and determine the instability of the rupture of the arterial plaque. Among several infectious microorganisms (Yuhua Sun, et al Atherosclosis, 2005, 179: 133. sup. 137), the most important is Chlamydia pneumoniae (Sun Yuhua, et al. J. Chinese medicine, 2004, 84: 1990. sup. 1993).

Chlamydia pneumoniae (Chlamydia pneumaniae, Cp, also abbreviated as c. pneumaniae, Cp or CPn in the literature) was the first Chlamydia isolated from the conjunctival secretion of the eyes of children in taiwan in 1965, when named tw (taiwan) -183, and in 1983, another Chlamydia isolated from the pharyngeal secretion of the university student with Acute respiratory infection in seattle in usa in 1, named AR-39 (act respiratory-39), and was identified by research.

Cp belongs to a special microorganism, is intermediate in size between bacteria and viruses, and is a common pathogen of respiratory tract infection. Cp has a unique growth cycle, with 2 tautomeric morphologic and functional phenotypes: the protomer and the provenance. The protomer is blocked in metabolism, is infectious and can survive outside cells for a short time; the primordia are states of intracellular Cp reproduction and are not infectious. Cp readily evades the host immune response and antibiotics also kill it poorly.

In 1988, Swedish scholars first reported that the positive rate of serum anti-Cp antibodies in patients with acute myocardial infarction and coronary heart disease was significantly higher than that in the control (Saikku P, et al. Lancet, 1988, ii: 983-. Since then, several cross-sectional serological studies have also demonstrated that Cp infection is associated with coronary heart disease (Yuhua Sun, et al. International Journal of Cardiology, 2004, 97: 199-. Many histological studies detect Cp pathogens at arterial plaques. Under the influence of different studies and different detection techniques, the positive rate of immunohistochemical Cp detection is about 33-71% (Caroline WATSON. clinical Science, 2008, 114: 509-. Animal experimental studies also suggest that repeated Cp infection promotes development of arterial plaque and is species-specific. Cp pathogens migrate from the lung into the normal or diseased arterial vessel wall by mononuclear macrophages and lymphocytes, persist in vascular endothelial cells, infect vascular smooth muscle cells, and participate in the inflammatory response of arterial plaque. Cp pathogens can be detected in peripheral blood, and the detection rate and the detection amount of Cp pathogen DNA in the arterial plaque are in positive correlation with the Cp pathogen DNA in the peripheral blood. However, prospective clinical studies of antibiotics for patients with coronary heart disease in European and American countries have failed to further confirm the relevance of Cp infection to coronary heart disease (Grayston JT, et al. N Engl J Med, 2005, 325: 1637-1645.Cannon CP, et al. N Engl J Med, 2005, 325: 1646-. The inventors believe that the design of those prospective clinical studies needs to be more rigorous and cannot rule out the role of Cp infection in the development of arterial plaque and related diseases. It is known that the mechanisms from development and rupture or erosion of arterial plaque, to local thrombosis, to the appearance of a range of clinical symptoms and the impact of various therapeutic measures on the prognosis of the associated disease are very complex (Sun Yuhua, Wang balding. perfusion, 2018, 33: 89-95).

At present, the theory of occurrence and development of arterial plaque and related diseases worldwide mainly lies in the cholesterol theory proposed by heart research of Foley Mingham in the United states 70 years ago, and the theory influences the prevention and treatment measures of the diseases related to the arterial plaque. In fact, compared with the history of more than 200 thousands years for human, the modern medicine can only start to recognize human health and diseases in 200 years. Cholesterol is a normal component of the human body and is a basic substance for human living metabolism, and if cholesterol is stared at, the problem of arterial plaque related diseases can be difficult to solve. In view of the severe situation of cardiovascular and cerebrovascular diseases affecting human health, new ideas and concepts are needed to induce new methods for detecting arterial plaque stability.

Disclosure of Invention

The invention firstly provides a novel method for detecting blood risk factors related to the occurrence and development of arterial plaque and the onset of cardiovascular and cerebrovascular diseases of an individual, which is characterized in that a Cp genome specific nucleic acid sequence is detected and the Cp infection state and the Cp infection degree are identified by testing peripheral venous whole blood, serum, plasma or separated mononuclear cells of an arterial plaque patient. If a Cp genome-specific nucleic acid sequence is detected, the Cp genome-specific nucleic acid sequence is used for indicating Cp pathogen infection in blood, indicating Cp infection in arterial plaque and being used as a blood risk factor for development of arterial plaque and related clinical diseases, particularly cardiovascular and cerebrovascular diseases.

Cp infection in the peripheral circulating blood of a human body indicates on the one hand Cp infection in arterial plaque and on the other hand Cp pathogens in the circulating blood bind to specific antibodies and also affect blood thrombosis. Specifically, when the levels of antigen and antibody are high, the amount of antibody binding to antigen to form a macromolecular immune complex is large, blood tends to be in a highly coagulated state or a pre-thrombotic state, and the risk of thrombus formation after rupture of arterial plaque is high. The antigen refers to a Cp pathogen or a metabolite thereof, and the specific anti-Cp antibody includes IgG and IgA. Therefore, the invention uses the detection of Cp pathogen infection in blood as a blood risk factor for the development of arterial plaque development and related diseases, particularly acute cardiovascular and cerebrovascular diseases.

In a second aspect of the invention, a method for detecting arterial plaque stability is provided. And (4) judging whether the detected arterial plaque is a bacterial arterial plaque or a sterile arterial plaque according to the fact that Cp infection exists or does not exist in the detected arterial plaque. That is, from the viewpoint of occurrence and development of an individual arterial plaque, the arterial plaque needs to be further classified. The occurrence and development of the bacterial arterial plaque are related to Cp infection, or named Cp infected arterial plaque, belong to unstable arterial plaque, are easy to rupture or erode, and cause the disease incidence related to the arterial plaque; the occurrence and development of sterile arterial plaque are not related to Cp infection, or are called Cp-free infected arterial plaque, belong to stable arterial plaque, and are not easy to generate plaque rupture or erosion and cause diseases related to arterial plaque. In other words, by detecting Cp infection in peripheral blood and arterial plaque, the arterial plaque of an individual can be classified into a bacterial arterial plaque or a sterile arterial plaque, and the stability of the arterial plaque of the individual can be further determined based on the classification.

Similar to other chronic infectious inflammations, in the bacterial arterial plaque, the inflammatory response of the arterial plaque is directly related to Cp infection, the infection degree and the related inflammatory response degree are continuously changed at different periods of inflammation, and the inflammatory response degree is also influenced by the immune response and disease resistance of an individual. For example, in certain stages of infectious inflammation, lesions may have a large number of pathogens of pathogenic microorganisms, while in other stages, lesions may have only a small number of pathogens, even no pathogens, and only a large number of fibrotic or calcified lesions, as is common in chronic infectious diseases. In the case of infectious inflammation, it is theorized that a small inflammatory response is self-healing.

Further, the invention also detects arterial plaques of different inflammatory stages of the individual. The inflammatory phase of the arterial plaque is staged according to whether Cp pathogens can be detected or not: there is Cp infection in the active or sterile inflammatory phase. Namely: in a certain period, Cp infection is detected in peripheral blood and in an arterial plaque, the arterial plaque is in a Cp infection inflammation activity period, and the larger the Cp infection amount is, the higher the inflammation activity degree is, the more easily the plaque is cracked or eroded, and the diseases related to the arterial plaque, particularly the acute cardiovascular and cerebrovascular diseases are caused; in another period, no Cp infection is detected in peripheral blood, indicating that no Cp infection is present in the arterial plaque, and the arterial plaque is in a sterile inflammation period, so that the plaque is stable and the risk of diseases related to the arterial plaque is low.

In conclusion, the stability of the individual arterial plaque is judged according to the Cp infection condition in the arterial plaque, and the inflammatory response activity of the arterial plaque is further staged according to the Cp infection degree. That is, the Cp infection in the arterial plaque belongs to unstable arterial plaque, and the more Cp pathogens, the more serious the inflammatory reaction, the more unstable the plaque, the more easily rupture or erosion, and the initiation of arterial plaque related diseases, especially acute cardiovascular and cerebrovascular diseases. The plaque is relatively stable without Cp infection in the arterial plaque.

Internationally, antibiotic administration was performed in patients with coronary heart disease for Cp infection, and the current administration was based on studies on specific anti-Cp antibody positivity in peripheral blood. Positive or high CpIgG antibody levels indicate past or recurrent Cp infection, which does not fully reflect the current Cp infection status and extent. The Cp genome-specific nucleic acid sequence in human peripheral blood is detected by PCR technology, and the current Cp infection state and infection degree in blood and arterial plaque of an individual can be shown.

In a final aspect of the invention, a method is provided for administering a suitable plaque-stabilizing or inflammatory response-reducing regimen as a result of the detected peripheral blood Cp infection status and extent of infection. Since patients with arterial plaque have Cp pathogens in their blood and, correspondingly, Cp pathogen infection in the arterial plaque, various regimens for the targeted application of inhibiting or killing Cp are contemplated, preferably azithromycin or roxithromycin. The medication regimen comprises regular, periodic dosing of azithromycin or roxithromycin for a period of time: regular and periodic dosing including daily, every other day oral or other routes of administration; the period of time is one to three weeks, or one to six months, etc. Dosing was continued regularly and periodically until the Cp pathogens were reduced or eliminated in the peripheral blood or arterial plaque.

Further, as a method for evaluating the effect of drug administration and for prognosis follow-up, it is necessary to periodically and repeatedly detect the infection status and infection degree of Cp pathogens in peripheral blood and in arterial plaque of a patient with arterial plaque, thereby judging the stability and inflammation status of arterial plaque, and determining whether to continue to carry out or adjust the drug administration regimen.

Preferably, the methods for detecting Cp pathogens in arterial plaque of a patient employed in the present invention include, but are not limited to, methods for detecting Cp genome-specific nucleic acid sequences in peripheral blood of a patient, indicative of Cp infection in the blood and in arterial plaque. Other methods, such as immunohistochemical techniques, also detect Cp infection in arterial plaque, are well known in the art.

The Cp genome-specific nucleic acid sequences include conserved sequences of the Cp inclusion body membrane protein coding gene (MOMP), which have specificity to distinguish other Chlamydia and are also commonly used for design and screening of primers and probes.

In some preferred embodiments, the presence of a pathogen Cp genome-specific nucleic acid sequence, a marker of Cp infection, can be determined by nucleic acid amplification techniques, such as general PCR. Variations of PCR include nested PCR and reverse transcriptase PCR. In certain embodiments, a wide variety of PCR variants and modifications can be used, such as real-time fluorescent quantitative PCR, the Taqman probe method commonly used for fluorescent quantitative PCR, which is well known in the art and can be used by one skilled in the art in practicing the present methods.

There are many methods for clinical detection of Cp infection, and the common techniques are real-time fluorescent quantitative PCR, and also normal temperature nucleic acid amplification techniques. Preferably, fluorescent qualitative or quantitative PCR techniques, including real-time fluorescent quantitative PCR techniques, have been established for detecting Cp genome-specific nucleic acid sequences in human peripheral blood mononuclear cells, which are well-known in the art as to their reproducibility, high specificity and high sensitivity. PCR reaction raw materials, reagents, instrumentation and rapid detection kits for Cp genome-specific nucleic acid sequence detection are commercially available.

Techniques for venous blood collection and isolation of mononuclear cells are well known in the art. The peripheral blood for Cp genome-specific nucleic acid sequence detection includes venous whole blood, serum, plasma or isolated mononuclear cells from human body.

Currently, the PCR technology for detecting Cp-specific nucleic acid sequences is widely used in clinical applications for respiratory infectious diseases.

The bacterial arterial plaque of the invention refers to the microbial infection in the arterial plaque, in particular to Cp pathogen infection, and the bacterial arterial plaque indicates that the arterial plaque is in a Cp infection inflammation stage, the more pathogens, the stronger inflammation reaction, the easy increase of plaque volume or the easy rupture erosion, and belongs to unstable arterial plaque.

The sterile arterial plaque is an arterial plaque without microbial infection in the plaque, has relatively weak inflammatory response and low risk of rupture, and belongs to a stable plaque.

The stability or instability of the arterial plaque mainly refers to the characteristics of whether the arterial plaque is cracked or eroded, and also includes the properties of whether the arterial plaque is developed, increased in volume and aggravated in the stenosis degree of the artery lumen.

The diseases caused by rupture or erosion of the arterial plaque can be serious acute cardiovascular and cerebrovascular diseases caused by related large vessel blockage, have clinical symptoms of typical acute cardiovascular and cerebrovascular diseases, can also be coronary microvascular myocardial infarction or cerebral infarction caused by microvascular embolism, and have only non-serious symptoms of transient chest pain, chest distress or syncope, amaurosis nigromaculata and the like. For those with arterial plaque, less severe symptoms may also be a precursor to the onset of severe acute cardiovascular and cerebrovascular disease.

It must be noted that not every development of arterial plaque, or every rupture and erosion of arterial plaque, leads to the appearance of clinical disease symptoms. Whether the rupture of the arterial plaque causes the onset of the relevant clinical disease is influenced by blood factors in addition to factors of the ruptured plaque, such as the size of the ruptured necrotic core, the position of the rupture opening, the amount of substances which are released into the blood to cause thrombosis, and the like.

The arterial plaque and related lesions related to the present invention also include atherosclerotic, calcified or non-calcified lesions, arterial luminal stenosis, etc. which are well known in the art, and are mainly the result of examining systemic arteries such as carotid artery, coronary artery, intracerebral artery, subclavian artery, upper and lower extremity arteries, aorta, etc. by applying techniques such as extravascular ultrasound, intravascular ultrasound, angiography, CT, optical coherence tomography, nuclear magnetic resonance, etc.

The indexes of the arterial plaque tending to be stable comprise plaque volume reduction, necrotic core reduction, arterial plaque disappearance, thin fiber plaque change into thick fiber plaque or fiber plaque, the degree of arterial lumen stenosis reduction, neovascularization reduction or disappearance in the arterial plaque and the like displayed by the inspection results; in contrast, indicators of arterial plaque instability include increased plaque volume, enlarged necrotic core, thinning of plaque fibroids, appearance or increase of neovascularization within the plaque, rupture or erosion of the plaque, increased stenosis of the arterial lumen, and the like.

The diseases related to the arterial plaque comprise coronary atherosclerotic plaque or stenosis, coronary heart disease, angina, acute myocardial infarction, old myocardial infarction, sudden death, coronary artery beading lesion or neoplasia expansion, aortic expansion or dissecting aneurysm, renal artery stenosis, iliac artery stenosis, upper limb or lower limb ischemia caused by artery plaque or lumen stenosis or blockage, carotid artery plaque and stenosis, subclavian artery stenosis, cerebrovascular stenosis, cerebral thrombosis, cerebral vascular rupture, hypertension and the like.

The invention relates to acute cardiovascular and cerebrovascular diseases related to arterial plaque, which comprise unstable angina, acute myocardial infarction, sudden cardiac death caused by coronary thrombosis, transient cerebral ischemia caused by cerebral thrombosis and cerebral apoplexy, wherein the cerebral apoplexy comprises cerebral thrombosis and cerebral vascular rupture caused by cerebral atherosclerosis rupture.

The index of the decrease in the incidence of the disease relating to arterial plaque according to the present invention includes improvement in the symptoms of the patient, decrease in the incidence of the disease, prolongation of the life expectancy, and the like.

The purpose of the invention is as follows: the new method for detecting the stability of the arterial plaque and the method adopting the corresponding medication scheme for stabilizing the arterial plaque and reducing the inflammatory response have positive effects on reducing the morbidity of diseases related to the arterial plaque, particularly acute cardiovascular and cerebrovascular diseases.

And (3) benefit analysis: the invention relates to a method for testing human peripheral blood, in particular to a method for testing the stability of arterial plaque, which is used for testing Cp infection in the peripheral blood and in the arterial plaque by testing Cp genome-specific nucleic acid sequences in the peripheral blood. The following three aspects are specified: cp infection in peripheral blood as a new blood risk factor for development of arterial plaque development and associated disease pathogenesis; the novel method for detecting the stability of the arterial plaque also classifies and stages the inflammatory response of the arterial plaque according to the Cp infection condition and the infection degree in the individual arterial plaque; determining the application method of the dosage scheme for stabilizing arterial plaque and reducing inflammatory response according to the detected Cp infection conditions in the peripheral blood and the arterial plaque of the human. The method provided by the invention is simple and effective, is noninvasive and harmless, is easy to copy, can be implemented in a large amount in a short time, and has good economic value prospect and good social value.

Detailed Description

The examples are intended to illustrate the invention, but not to limit the scope of the invention. The disease examination methods and experimental techniques described in the examples are conventional means well known to those skilled in the art, and the equipment and detection reagents used are commercially available.

Example collection: 40 carotid ultrasonic inspectors in Beijing, a physical examination center were collected, and 23 males aged 53 years. Of these, 10 cases were with carotid plaque or luminal stenosis and a history of old myocardial infarction, 19 cases were with carotid plaque and carotid stenosis, and 11 cases were without carotid plaque. The basic characteristics and carotid artery test results of the three groups of examples are shown in Table 1.

Carotid artery ultrasonic examination: carotid artery ultrasonic examination can be used as a window index for measuring whether arterial plaque is developed and the severity of the arterial plaque in systemic aorta blood vessels, particularly coronary arteries and cerebral arteries, has the characteristics and advantages of rapidness, economy, effectiveness, no wound and no harm, and is often used as a consideration index in the field of arterial plaque research. The present embodiment uses ultrasound inspection of carotid arteries and image results as a method for detecting arterial plaque and assessing its stability. The carotid artery ultrasonic examination adopts a domestic color ultrasonic diagnostic apparatus, the frequency of the probe is 5-12MHz linear array probe, and a gray scale imaging mode is adopted. The right side of the probe starts from the bifurcation of the innominate artery and the left side of the probe starts from the beginning of the main artery arch, and the main artery and branches of the common carotid artery, the bifurcation of the internal and external carotid arteries, the internal carotid artery and the external carotid artery are continuously probed by scanning in a transverse cutting mode and a longitudinal cutting mode (including an anterior position, a posterior position, an internal position, an external position and an anterior position). The carotid intima thickness is 1.0-1.5mm, which is the intimal thickening. The thickness of the inner membrane is more than or equal to 1.5mm or the thickness of the limited inner membrane is more than 50% of that of the peripheral inner membrane, and then the plaque is formed. The examination indexes include the thickness of the blood vessel intima, whether plaque exists, the plaque distinguishes soft plaque, hard plaque and mixed plaque, the degree of stenosis of the artery lumen, whether plaque is cracked or ulcerated, whether plaque surface is smooth or thrombus exists, and the like.

Peripheral venous blood collection and mononuclear cell DNA extraction: respectively collecting 5ml of peripheral venous blood, performing EDTA anticoagulation, conventionally separating mononuclear cells, extracting DNA according to the kit instructions, storing at-20 ℃ and performing unified preparation and detection.

And (3) PCR detection: this example employs the fluorescent quantitative PCR detection technique Cp infection. The kit is characterized in that a domestic real-time fluorescent quantitative PCR instrument and a commercial Chlamydia pneumoniae probe method nucleic acid quantitative detection kit are used, and an upstream primer sequence: 5'-AGTTGAGCATATTCGTGAGG-3', downstream primer sequence: 5'-TTTATTTCCGTGTCGTCCAG-3', Taqman fluorescent probe sequence: FAM-5'-ACAACCATCAGTATCTCACAAGGCAACAC-3' BHQ1, FAM is a fluorescence reporter group, and BHQ1 is a fluorescence quenching group. The recombinant plasmid carries a species-specific chlamydia pneumoniae Pst I fragment as a standard. A20. mu.l PCR reaction system containing about 150ng of DNA was set up, and 2 parts of a negative control (ultrapure water), 2 parts of a 10-fold gradient diluted standard sample, and 2 parts of each DNA sample to be tested were set up. Real-time fluorescent quantitative PCR amplification procedure: 5min at 95 ℃; 30s at 95 ℃; 60 ℃ for 60s, 40 cycles of amplification, and reading the fluorescence signal at 60 ℃ annealing for each cycle. And (4) judging a result: the amplification curve is an s-shaped curve, and if the Ct value is less than 30, the amplification curve is judged to be positive; if the Ct value is more than or equal to 30 and less than 40, the positive is weak; if the Ct value is greater than 40, the result is judged to be negative.

Cp infection detection results: the total Cp infection positive rate was 50% for 40 subjects. Of 29 carotid plaque patients, 19 peripheral blood patients had Cp infection, with a positive rate of 65.5% (19/29), average: 1607 copies/10⁴Peripheral blood mononuclear cells, range: 102-17211 copy/10⁴Peripheral blood mononuclear cells. Of 11 patients without plaque in the carotid artery, only 1 of them showed a weak positive Cp infection (56 copies/10)⁴Peripheral blood mononuclear cells).

Partial individual example cases

Example 1, no dyslipidemia, no hypertension, no diabetes, no plaque in carotid, and negative Cp infection.

Example 2, hypertension, diabetes, carotid plaque, fibrotic plaque or calcified hard plaque, no rupture, Cp infection negative, sterile arterial plaque, belonging to stable plaque.

Example 3 dyslipidemia, Mixed plaques, rupture, Cp infection Positive, Cp nucleiAcid 7689 copies/10⁴Peripheral blood mononuclear cells, having bacterial arterial plaque, belonging to unstable arterial plaque, are recommended to implement a regimen against Cp infection.

Example 4 old myocardial infarction, carotid multiple plaque, rupture, Cp infection, Cp nucleic acid 17211 copies/10⁴Peripheral blood mononuclear cells, having bacterial arterial plaque, belonging to unstable arterial plaque, are recommended to implement a regimen against Cp infection.

Example 5 carotid stenosis > 70%, mixed plaques, ruptured, ulcerated and mural thrombus, Cp infection positive, Cp nucleic acid 10588 copies/10⁴Peripheral blood mononuclear cells, which are active in inflammatory phase with bacterial infection and belong to unstable arterial plaque, are recommended to implement a medication scheme aiming at Cp infection.

Example 6, diabetes, carotid plaque, > 70% carotid stenosis, fibrotic plaque or calcified hard plaque, no rupture, thrombus, Cp infection negative, sterile arterial plaque, belonging to stable plaque.

The above examples show the feasibility and effectiveness of the method for detecting the stability of arterial plaque provided by the invention, wherein the proposal of the method for Cp infection medication scheme is only used for reference, and whether to implement and how to implement the method in clinical practice needs to be determined by a clinician according to the individual situation by comprehensively measuring the advantages and disadvantages of the clinician.

TABLE 1 basic characteristics of three groups of examples and Chlamydia pneumoniae infection by carotid ultrasound and fluorescence quantitative PCR detection

Claims

1. A method for detecting the stability of arterial plaque of a human body is to detect chlamydia pneumoniae infection in the arterial plaque to judge the stability of the arterial plaque of the human body.

2. The method of claim 1, wherein the detecting chlamydia pneumoniae infection in arterial plaque is detecting chlamydia pneumoniae genome-specific nucleic acid sequences in peripheral blood of a human, indicating blood and intra-arterial plaque chlamydia pneumoniae infection.

3. The method of claim 2, wherein the nucleic acid sequence specific for the Chlamydia pneumoniae genome comprises a conserved sequence of Chlamydia pneumoniae genome inclusion body membrane protein genes.

4. The method according to claim 2, wherein the detection method of the Chlamydia pneumoniae genome-specific nucleic acid sequence in human peripheral blood comprises using PCR, fluorescence qualitative PCR, fluorescence quantitative PCR or the like.

5. The method of claim 4, wherein the fluorescent quantitative PCR technology is used for detecting the Chlamydia pneumoniae genome-specific nucleic acid sequence in human peripheral blood by detecting the copy number of the nucleic acid sequence.

6. The method according to claim 2, wherein the nucleic acid sequence specific for the Chlamydia pneumoniae genome is detected in human peripheral blood, wherein the peripheral blood comprises extracted venous whole blood, serum, plasma or isolated mononuclear cells.

7. A method of classifying arterial plaque in an individual by classifying arterial plaque of the individual according to claim 1 with or without chlamydia pneumoniae infection in the arterial plaque as a bacterial arterial plaque and without chlamydia pneumoniae infection in the arterial plaque as a sterile arterial plaque.

8. A method of classifying the stability of an arterial plaque of an individual by classifying the stability of an arterial plaque according to the bacterial or sterile arterial plaque of claim 7, the bacterial arterial plaque being an unstable arterial plaque and the sterile arterial plaque being a stable arterial plaque.

9. A method for detecting a blood risk factor for the onset of an arterial plaque-related disease in an individual, said method comprising determining the risk of the onset of an arterial plaque-related disease by detecting Chlamydia pneumoniae infection in blood according to claim 2.

10. A method for detecting the stability of an arterial plaque of an individual, wherein the method is used for detecting the stability of the arterial plaque according to the detection condition of chlamydia pneumoniae infection in the arterial plaque, such as chlamydia pneumoniae infection in the arterial plaque, namely bacterial arterial plaque, belongs to unstable plaque, is easy to crack or erode, and causes diseases related to the arterial plaque to develop; if the artery plaque is free from chlamydia pneumoniae infection, the artery plaque is a sterile artery plaque, belongs to a stable plaque, and is not easy to crack or erode the plaque and cause diseases related to the artery plaque.

11. A method for detecting the active period of the bacterial and inflammatory diseases of the arterial plaque of an individual, which is characterized in that the active period or the sterile inflammatory period of the arterial plaque is judged by detecting the infection condition of the chlamydia pneumoniae in the arterial plaque according to claim 1, if the chlamydia pneumoniae infection exists in the arterial plaque, the arterial plaque is in the active period of the bacterial and inflammatory diseases, and diseases related to the arterial plaque are easy to develop; if the artery plaque is not infected by chlamydia pneumoniae, the artery plaque is in an aseptic inflammation stage, and the plaque is relatively stable and is not easy to cause diseases related to the artery plaque.

12. A method for detecting the infection degree and inflammation activity degree of chlamydia pneumoniae of an arterial plaque of an individual, which comprises judging the infection degree and inflammation activity degree of chlamydia pneumoniae of the arterial plaque according to the copy number of the detection nucleic acid sequence in claim 5, indicating the rupture or erosion risk of the plaque and the risk of thrombosis and diseases related to the arterial plaque.

13. A method of determining an individual to take a regimen for stabilizing arterial plaque and reducing inflammatory response by detecting chlamydia pneumoniae infection in blood and in arterial plaque according to claim 2, the regimen comprising the use of azithromycin or roxithromycin or the like, for the targeted use of inhibiting or killing chlamydia pneumoniae pathogens.

14. A method of determining whether the subject's regimen for stabilizing arterial plaque and reducing inflammatory response according to claim 13 is to be continued by detecting whether there is any chlamydia pneumoniae infection in the blood and in the arterial plaque of the subject according to claim 2.

15. The method according to any one of claims 9-14, wherein the disease is an acute cardiovascular disease.