Method of neurosyphilis diagnostics

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.

4 dwg, 3 ex

 

The invention relates to medicine, namely to laboratory diagnosis, and can be used in determining the severity of lesions of brain structures pale pallidum in patients with syphilis in the anamnesis.

The problem of timely diagnosis of neurosyphilis remains relevant despite the creation of a broad base of laboratory tests, including clinical and serological indices in the study of cerebrospinal fluid. However, due to insufficient sensitivity of tests used often situations arise in which we cannot exclude or confirm the invasion of Treponema pallidum in the structure of the nervous system. In some cases the skill of the Clinician is crucial in the diagnosis of this pathology, and the timely appointment of adequate therapy helps prevent irreversible organic disorders of the nervous system and death of the patient. At the same time, there are some forms of early asymptomatic neurosyphilis, i.e. no clinical signs of disease with loss of sensitivity to traditional laboratory methods and follow.

Known method for the diagnosis of neurosyphilis in the study of cerebrospinal fluid by using serological tests (Lukyanov A. M. Neurosyphilis. Modern aspects of the clinic, diagnosis, therapy. Minsk, Paradox, 2009, pp. 246-263),�Lucasi definition and research VDRL (Veneral Disease Reserch Laboratory), TPHA, Rifts (REEF with whole spinal fluid, ELISA-IgG.

Currently, however, the results of serological tests in the study of cerebrospinal fluid in most cases, are insensitive or low-sensitive, which makes the diagnosis of neurosyphilis. Such situation has developed in the 21st century in connection with the change in the response of humans to the widespread use of antibiotics, which was not mentioned earlier (first half of the 20th century) when the above tests were highly sensitive and definitely solved the problems of diagnosis of neurosyphilis.

Known method for the diagnosis of neurosyphilis (Patent RF №2230323, IPC 33/50, publ. 2004), including the identification of sphingomyelin in the cerebrospinal fluid of patients with flow-through thin-layer chromatography, the level of which diagnosis is performed.

The disadvantage of this method is its complexity and time consuming, require expensive equipment and reagents, and the availability of highly qualified personnel. This method can be carried out only in specialized laboratories of the research Institute, so in practice it is difficult to use it.

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

The disadvantage of this method is that currently the body's sensitivity to inflammatory processes caused by the introduction of Treponema pallidum in the brain structure, decreased sharply due to the wide use by the population antibacterial drugs that have influenced a decrease in the immune response to a foreign antigen.

In connection with this object of the invention is to eliminate these drawbacks, and to provide objective criteria for the diagnosis of neurosyphilis, early, including preclinical diagnosis of the disease, reducing the time of data acquisition, reduction in the cost of conducting research.

For this method, the diagnosis of neurosyphilis, including microscopic examination of cerebrospinal fluid specimens, it is suggested to marginal dehydration of samples of cerebrospinal fluid and microscopic examination of exercise in polarized light. Thus the detection of anisotropic structures in the form of dendrites or spherulites, which are located inside of the formations in the shape of ovals, containing lipids, are diagnosed with an early form of neurosyphilis; in identifying the many ovals that are grouped in the form of balls included in the anisotropic structure and/or separately located, d�Instituut a late neurosyphilis.

It is known that the liquid crystal composition of the structures based on the "lipid - water", "lipid - protein - water", which when translated in solid state the method of boundary dehydration of biological fluids form the structure observed by microscopy in polarized light (Shabalin V. N., Shatokhina S. N. The morphology of biological fluids of man. M., Triad, 2001, p. 52-54). The cell membrane of the brain and spinal cord is rich in lipids and proteins. When the destruction of degradation products into the cerebrospinal fluid, one function of which is to eliminate toxins from the brain tissue. As Treponema pallidum penetrates into the brain and forms in it multiple foci of destruction, causing pathological changes in the Central nervous system (paresis, paralysis, stroke, mental disorders, etc.), the idea is to explore patterns of cerebrospinal fluid systems "lipid - water", "lipid - protein - water" in patients who have had syphilis, for the purpose of diagnosis of destructive processes in the brain tissue.

Fig. 1 shows the spherulite cerebrospinal fluid of a healthy person. It is seen that the structure is monolithic and lacks any additional inclusions (×300); Fig. 2 shows the anisotropic dendrite with the presence of inclusions in the form of ovals containing the lipids in ill�with early asymptomatic neurosyphilis. Amorphous ovals characterize the processes of destruction in the Central nervous system (×300); Fig. 3 - lots of ovals, grouped in the form of balls located inside the spherulite and outside (×50) of Fig. 4 - the same (×200). The method is as follows.

The patient, a history which was syphilis (seropositive reaction serum for syphilis), receive cerebrospinal fluid for diagnosis of neurosyphilis. To do this, conduct a study on the method of boundary dehydration, which on the surface of a glass slide put a drop of cerebrospinal fluid and cover it with a cover glass (creation analysis cell). After 72 hours the cells mikroskopiruût in polarized light at various magnifications and the detection of structures is diagnosed early or late form of neurosyphilis.

Example 1

Patient M., 36 years old. 2 years ago diagnosed with syphilis. Received a course of specific therapy. Currently the patient is in a psychiatric hospital, inadequate, disoriented in space. The results of clinical studies of the cerebrospinal fluid: the concentration of protein - 0,47; cytosis 5/3 (lymphocytes - 3, monocytes - 2), which corresponds to the parameters of normal.

Serology: VDRL - negative, TPHA - weakly positive (2+), Rift - positive (3+), ELISA G - negative�, i.e. the outcome is uncertain.

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

Conclusions: neurosyphilis, early form.

The examining Clinician: there are medical history and neuropsychological data, which together with the results obtained by the claimed method, allow the diagnosis of neurosyphilis and prescribe a specific course of therapy for neurosyphilis.

It was followed by adequate treatment, after 6 months re-examination of the cerebrospinal fluid by the proposed method did not reveal structures characteristic of neurosyphilis.

Example 2

Patient F., 52 years. Installed late syphilis. A course of specific therapy not received. Currently enrolled in the Department of neurology at the effects of cerebral infarction, right-sided hemiparesis.

The results of clinical studies of the cerebrospinal fluid: the concentration of protein - 0,21; cytosis 8/3 (lymphocytes - 8), which corresponds to the parameters of normal.

Serology: VDRL - negative, TPHA - weakly positive (2+), Rift - weakly positive (2+), ELISA G is weakly positive (R=2,5), i.e. the outcome is uncertain.

The study results of the claimed method: the analytical cell was found a large number of ovals that are grouped�'s in the form of balls, included in anisotropic structures and outside them (Fig. 3, 4). Conclusions: neurosyphilis, late meningovascular form.

The examining Clinician: there is neurological data together with the results obtained by the claimed method, allow the diagnosis of late forms of neurosyphilis and prescribe a specific course of therapy for neurosyphilis.

Example. 3

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

The study carried out by the above procedure.

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

Conclusion: signs of neurosyphilis are missing.

The examining Clinician: clinical signs of neurosyphilis is not found.

The proposed method studies have been conducted among 106 people, with a history of syphilis and various neurological symptoms, was obtained cerebrospinal fluid for diagnosis of neurosyphilis.

Studies have been carried out by known methods: neurosyphilis was confirmed by serological studies and the results of the cell count of the cerebrospinal fluid in 42 people. The remaining 64 patients were obtained negative or weakly positive results.

Those patients (64 men), which demanded�camping further diagnosis, a study was conducted of the claimed method. The results showed that the destructive process in the Central nervous system caused pallidum pale, had only 26 people. These patients were assigned to additional treatment and at the follow-up study after 6 months was noted positive dynamics of neurological symptoms (recovery of speech, sensation, motor function) and no signs of neurosyphilis claimed method when you study. The remaining 38 patients, the diagnosis of neurosyphilis was withdrawn.

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 the structures of the brain that allows you to confidently make the diagnosis of neurosyphilis and promptly appoint a specific therapy. In sbeh MOCVD this method demanded by clinicians, especially in those cases where the diagnosis of neurosyphilis is controversial. The modern diagnostics provides targeted and therefore effective therapy.

Method for the diagnosis of neurosyphilis, including microscopic examination of samples of cerebrospinal fluid, characterized in that the conducting boundary dehydration of the samples spinnomozgova� fluid, and their microscopic examination is carried out in polarized light and the detection of anisotropic structures in the form of dendrites or spherulites, which are located inside of the formations in the shape of ovals, containing lipids, are diagnosed with an early form of neurosyphilis, and in identifying the many ovals that are grouped in the form of balls included in the anisotropic structure and/or separately located, are diagnosed with a late neurosyphilis.



 

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