RU2554765C1 - Method of neurosyphilis diagnostics - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method of neurosyphilis diagnostics includes the microscopic analysis of cerebrospinal fluid samples, with carrying out edge dehydration of the cerebrospinal fluid samples, and their microscopic analysis being carried out in a polarised light; in case of the detection of anisotropic structures in the form of dendrites or spherulites inside which oval-shaped formations, containing lipids, are located, an early form of neurosyphilis is diagnosed; and in case of the detection of a multitude of ovals, aggregated in the form of balls, included in the anisotropic structures and/or located separately, late meningovascular neurosyphilis is diagnosed. The invention task is to obtain objective criteria for the diagnostics of neurosyphilis, provision of early, including pre-clinical diagnostics of the disease, reduction of the terms for obtaining data, reduction of costs for carrying out analyses.

EFFECT: in a number of cases the method is the only one which makes it possible to obtain the valid criteria for diagnostics in the form of a picture of the brain structures destruction, which makes it possible to diagnose neurosiphilis confidently and prescribe specific therapy in due time.

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Description

The invention relates to medicine, namely to laboratory diagnostics, and can be used to determine the severity of damage to brain structures of pale treponema in patients with a history of syphilis.

The problem of timely diagnosis of neurosyphilis remains relevant, despite the creation of a wide base of laboratory tests, including general clinical and serological parameters in the study of cerebrospinal fluid. However, due to the insufficient sensitivity of the tests used, situations often arise in which the invasion of pale treponema into the structures of the nervous system cannot be ruled out or confirmed. In some cases, only the clinician's experience is crucial in the diagnosis of this pathology, and the timely appointment of adequate therapy helps to prevent irreversible organic disorders in the nervous system and the death of the patient. At the same time, there are forms of an early asymptomatic course of neurosyphilis, i.e. there are no clinical signs of the disease with loss of sensitivity of traditional laboratory methods and follow-up.

A known method for the diagnosis of neurosyphilis in the study of cerebrospinal fluid using serological tests (Lukyanov AM Neurosyphilis. Modern aspects of the clinic, diagnosis, therapy. Minsk, Paradox, 2009, p. 246-263), including the definition and study of VDRL (Veneral Disease Reserch Laboratory), RPGA, RIFc (RIF with whole cerebrospinal fluid), ELISA-IgG.

However, at present, the results of serological tests in the study of cerebrospinal fluid in most cases are insensitive or insensitive, which does not allow the diagnosis of neurosyphilis. This situation developed in the 21st century due to a change in the reaction of the human body to the widespread use of antibiotics, which was not noted earlier (the first half of the 20th century), when the above tests were highly sensitive and unequivocally addressed the diagnosis of neurosyphilis.

A known method for the diagnosis of neurosyphilis (RF Patent No. 2230323, IPC 33/50, publ. 2004), including the determination of sphingomyelin in the cerebrospinal fluid of patients using flow thin-layer chromatography, the level of which is diagnosed.

The disadvantage of this method is its complexity and complexity, requires expensive equipment and reagents, as well as the presence of highly qualified personnel. This method can only be carried out in the laboratories of specialized research institutes, so it is difficult to use it in practical work.

Closest to the claimed invention is a method for the diagnosis of neurosyphilis, including microscopic examination of cerebrospinal fluid (Lukyanov A.M. Neurosyphilis. Modern aspects of the clinic, diagnosis, therapy. Minsk, Paradox, 2009, p. 233).

The disadvantage of this method is that at present the sensitivity of the body to inflammatory processes caused by the introduction of pale treponema into the brain structures has sharply decreased due to the widespread use of antibacterial drugs by the population, which has affected the decrease in the body's immune response to a foreign antigen.

In this regard, the object of the invention is to remedy these disadvantages, obtain objective criteria for the diagnosis of neurosyphilis, provide early, including preclinical diagnosis of the disease, reduce the time required to obtain data, reduce research costs.

To this end, in a method for the diagnosis of neurosyphilis, including a microscopic examination of cerebrospinal fluid samples, it is proposed to carry out regional dehydration of cerebrospinal fluid samples, and to carry out their microscopic examination in polarized light. In this case, when revealing anisotropic structures in the form of dendrites or spherulites, within which formations in the form of ovals containing lipids are located, an early form of neurosyphilis is diagnosed; when revealing many ovals, grouped in the form of balls included in anisotropic structures and / or separately located, late meningovascular neurosyphilis is diagnosed.

It is known that the composition of liquid crystal structures is based on the systems “lipid-water”, “lipid-protein-water”, which, when transferred to the solid-state state by the method of edge dehydration of biological fluids, form the structures observed by microscopy in polarized light (Shabalin V.N. Shatokhina S.N. Morphology of human biological fluids. M., Triad, 2001, p. 52-54). The membranes of the cells of the brain and spinal cord are rich in lipids and proteins. When they are destroyed, degradation products enter the cerebrospinal fluid, one of the functions of which is the removal of toxic substances from brain tissue. Since pale treponema penetrates the brain and forms multiple foci of destruction in it, causing pathological changes in the central nervous system (paresis, paralysis, strokes, mental disorders, etc.), an idea arose - to study the structures of the cerebrospinal fluid of the lipid-water systems, "Lipid - protein - water" in patients who have suffered syphilis, in order to diagnose destructive processes in brain tissue.

In FIG. 1 shows spherulitis of cerebrospinal fluid of a healthy person. It can be seen that its structure is monolithic and it does not contain any additional inclusions (× 300); in FIG. Figure 2 shows an anisotropic dendrite with inclusions in the form of ovals containing lipids in a patient with early asymptomatic neurosyphilis. Amorphous ovals characterize the processes of destruction in the central nervous system (× 300); in FIG. 3 - a plurality of ovals grouped in the shape of balls located inside and outside the spherulite (× 50), in FIG. 4 - the same (× 200). The method is as follows.

A patient with a history of syphilis (seropositive reaction of blood serum to syphilis) is given cerebrospinal fluid to diagnose neurosyphilis. To do this, conduct a study by the method of regional dehydration, for which a drop of cerebrospinal fluid is applied to the surface of a glass slide and covered with a coverslip (creating an analytical cell). After 72 hours, the cells are microscopied in polarized light at various magnifications and, if structures are detected, an early or late form of neurosyphilis is diagnosed.

Example 1

Patient M., 36 years old. 2 years ago, a syphilis diagnosis was made. Got a course of specific therapy. Currently, the patient is in a psychiatric hospital, inadequate, disoriented in space. The results of a general clinical study of cerebrospinal fluid: protein concentration - 0.47; 5/3 cytosis (lymphocytes - 3, monocytes - 2), which corresponds to normal parameters.

Serological tests: VDRL - negative, RPHA - slightly positive (2+), RIFc - positive (3+), ELISA G - negative, i.e. the result is doubtful.

The results of the study of the claimed method: dendrites with ovals included in their structure were detected (Fig. 2).

Conclusion: neurosyphilis, early form.

Clinician examination: there are anamnestic and neuropsychiatric data, which, together with the results obtained by the claimed method, make it possible to diagnose neurosyphilis and prescribe a course of specific therapy for neurosyphilis.

Adequate treatment was carried out, after 6 months a repeated examination of the cerebrospinal fluid by the proposed method did not reveal structures characteristic of neurosyphilis.

Example 2

Patient F., 52 years old. Established late syphilis. I did not receive a course of specific therapy. Currently, she entered the neurological department about the consequences of cerebral infarction, right-sided hemiparesis.

The results of a general clinical study of cerebrospinal fluid: protein concentration - 0.21; cytosis 8/3 (lymphocytes - 8), which corresponds to the parameters of the norm.

Serological studies: VDRL - negative, RPHA - weakly positive (2+), RIFc - weakly positive (2+), ELISA G - weakly positive (KP = 2.5), i.e. the result is doubtful.

The results of the study of the claimed method: in the analytical cells found a large number of ovals, grouped in the form of balls included in anisotropic structures and outside them (Fig. 3, 4). Conclusion: neurosyphilis, late meningovascular form.

Clinician examination: there are neurological data that, together with the results obtained by the claimed method, make it possible to diagnose the late form of neurosyphilis and prescribe a course of specific therapy for neurosyphilis.

Example. 3

Patient O., 40 years old. Moved syphilis 8 years ago. Got 2 courses of specific therapy.

A study was carried out according to the method described above.

The results of the study of the claimed method: in all analytical cells detected spherulites with normal parameters (Fig. 1).

Conclusion: there are no signs of neurosyphilis.

Clinician examination: no clinical signs of neurosyphilis were detected.

According to the proposed method, studies were conducted in 106 people with a history of syphilis and various neurological symptoms, cerebrospinal fluid was obtained to diagnose neurosyphilis.

Studies were carried out by known methods: neurosyphilis was confirmed by serological studies and the results of counting cerebrospinal fluid cells in 42 people. The remaining 64 patients received negative or weakly positive test results.

For those patients (64 people) who needed to clarify the diagnosis, the study was conducted by the claimed method. The results showed that the destructive process in the central nervous system caused by pale treponema occurred in only 26 people. These patients were prescribed additional treatment and, after a follow-up study, after 6 months, there was a positive dynamics of neurological symptoms (restoration of speech, sensitivity, motor functions) and the absence of signs of neurosyphilis according to the claimed method with repeated examination. The remaining 38 patients were diagnosed with neurosyphilis.

The claimed method for the diagnosis of neurosyphilis in some cases is the only one that allows to obtain reasonable diagnostic criteria in the form of a picture of the destruction of brain structures, which allows you to confidently diagnose neurosyphilis and timely prescribe specific therapy. In GBOZ MO MOKKVD this method is in demand by clinicians, especially in cases where the diagnosis of neurosyphilis is controversial. Conducting timely diagnosis provides targeted and, therefore, effective therapy.

Claims (1)

A method for diagnosing neurosyphilis, including microscopic examination of cerebrospinal fluid samples, characterized in that they conduct regional dehydration of cerebrospinal fluid samples, and their microscopic examination is carried out in polarized light and when anisotropic structures are detected in the form of dendrites or spherulites, inside of which there are formations in the form of ovals containing lipids, diagnose an early form of neurosyphilis, and when revealing many ovals, grouped in the form of balls included in the anisotra optic structures and / or separately located, diagnose late meningovascular neurosyphilis.

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