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Method for predicting the character of bacterial keratitis flow

Method for predicting the character of bacterial keratitis flow
IPC classes for russian patent Method for predicting the character of bacterial keratitis flow (RU 2245553):

G01N33/68 - involving proteins, peptides or amino acids
G01N33/535 -
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FIELD: medicine, ophthalmology.

SUBSTANCE: in lacrimal liquid one should detect the content of interleukin 8 (IL-8) and that of interleukin 1 beta (IL-1β) to calculate prognostic coefficient (PC) due to dividing the first value by the second one by the following formula: At PC value being below 10.0 one should predict favorable disease flow, and at PC value being above 10.0 - unfavorable flow.

EFFECT: higher accuracy of prediction.

2 ex

 

The invention relates to medicine, namely to ophthalmology, and can be used as a method of predicting the nature of the flow in bacterial keratitis.

Bacterial keratitis is one of the common causes of sudden vision loss, blindness and eye loss. In this regard, the prediction of the occurrence of the complicated course of the disease early in the disease will be of great help for the appointment of adequate treatment from the first day.

Known factors that contribute to the emergence of bacterial keratitis: corneal injury and subsequent merger of purulent infection, soft contact lenses, chronic conjunctivitis, purulent actionistic, irrational use of antibiotics and corticosteroids (Michael E, Gorgiladze M.S. Application acterized in combination with pulsed electromagnetic field in the treatment of bacterial keratitis // Ophthalmological journal, 1993, No. 3, pp.163-165; Marchuk Û.F. Eye infection // Russian medical journal, in 1999, v.7, №1, - p.16-19).

However, these factors were used only to determine the causes of ulcers of the cornea, without considering the possible relationship of these data to predict future disease: favorable (epithelialization of the ulcer) or complicated course (abscess of the cornea, corneal perforation, endophthalmitis, pan is talmit).

Known methods of diagnosis of severe adverse currents of bacterial keratitis in immunological parameters of blood on the basis of immunodeficiency on T - and b-systems of immunity (Silchenko T.S., Sakovich V.N., Immunoreactivity with suppurative keratitis. // Ophthalmological journal, 1990, N 7, s-411).

However, a disadvantage of this method of diagnosis is a statement of changes only systemic immunological parameters without reflection condition of the local resistance, namely in the tear fluid. It should be noted that changes in systemic immunity can be, and other accompanying diseases.

Closest to the claimed method and selected as a prototype is a method for predicting the effectiveness of treatment of bacterial keratitis, depending on the size of the ulcers and the use of corticosteroids, in which it is proved that the size of the defect of the cornea (4,4±2.4 mm) and the original corticosteroids are prognostic factors for treatment failure (see R.Y.Kim., .L.Cooper, L.D.Kelly “Prognostic factors for the efficacy of treatment of ulcerative bacterial keratitis”, 1996).

For reasons that impede the achievement of specified following technical result include insufficient information, and the accuracy of this method for diagnosis and prognosis is of aracter course of the disease.

The present invention is the prediction of the shape of the flow of bacterial keratitis: favorable with mild infections, cleansing and epithelialization of the ulcer focus and complicated course of the disease when activated purulent infection.

The technical result of the invention is determining the ratio of immunological parameters tear fluid characteristic favorable and complicated course of bacterial keratitis.

The problem is solved by examining the tear fluid, where, according to the invention, determine the content of interleukin 8 (IL-8) and the content of interleukin 1 beta (1β)hope prognostic factor (PC) by dividing the first figure by the second formula

and largest PC below 10,0 predicts favorable over and above 10.0 - complicated.

Studies on patent and scientific and technical information showed that the proposed method of forecasting is not known and should not be explicitly studied the prior art, i.e. meets the criteria of “novelty” and “inventive step”.

The method is as follows:

The definition of interleukines in the tear fluid is conducted by a method enzyme immunoassay using the test system is m “Cytokine” (S.-Petersburg). These test systems are based on the “sandwich”method, enzyme-linked immunosorbent assay with the use of horseradish peroxidase as indicator enzyme.

To determine 1L-1β take the tablets, and in the first two vertical rows of cells made 100 ál of standards analyzed cytokine: A - 0 PG/ml, 50 PG/ml, 250 PG/ml, D - 500 PG/ml, E - 1000 PG/ml, 2000 PG/ml In the remaining cells made of 100 ál of the samples. Samples and standards applied in the recommended buffers. Tablet incubare for 1.5 hours at 18-20°C. After incubation, the solution of the cells removed by pipette. Then the cells washed three times making 300 ál of wash solution to each of them. Second MCAT labeled with Biotin, made of 100 μl and inquirees for 1.5 hours with continuous shaking at 18°Spoke incubation solution from cells removed by pipette. Cells washed three times making 300 ál of wash solution to each of them. Conjugate with streptavidin horseradish peroxidase, diluted 1:100 with buffer made of 100 ál to all wells of the tablet and inquirees at 18°C with continuous shaking. After incubation, the solution of the cells removed by pipette. After that, carry out the steps of washing and dyeing.

To determine 1L-8, the first antibody, diluted planting buffer 1:5000 in 50 µl immobilized on the inner surface the parts of cells flexible tablets for ELISA.

After immobilization of the antibody solution was removed and cells washed twice. Into each hole to be made in 100 μl of blocking solution. Tablet incubare 1 hour at room temperature and continuous shaking. In the first two horizontal rows of cells made 50 ál of standards: AV - 0 PG/ml, 1L-8, A2B2 - 35 PG/ml, AV - 75 PG/ml, AV - 150 PG/ml, AV - 300 PG/ml, AV - 600 PG/ml, AV - 1.2 ng/ml, AV - 2.5 ng/ml, IV - 5 ng/ml, 1L-8. The remaining cells to be made in 50 ál of the samples. After incubation, the solution of cells is removed and washed twice. The second antibody 1L-8 in a dilution of 1:500 contribute 50 MLK and inquirees samples with them for 1 hour with continuous shaking. Third antibody diluted 1:50 and add 50 ál to well. Incubare tablet 1 hour at room temperature with continuous shaking. After this, carry out the steps of washing and dyeing.

See clinical studies have allowed us to distinguish the 2 forms of current bacterial keratitis:

1. favorable with mild infection, the solace of the eyeball, the cleansing of the ulcer bottom and epithelialization of the ulcer;

2. adverse with further activation of infection, low regeneration ability of the cornea, which leads to a sharp deterioration of vision and even blindness.

Each patient were identified prognostic factors, the range of which was from 8.0 to 16.6. With ateisticheskogo analysis and correlation revealed the dependence of the shape of the flow of bacterial keratitis on the magnitude of the prognostic factor.

Thus, PC1 from 8.3±0,2 9,6±0,3 shows a favorable course and gives the opportunity to predict 1 the flow of suppurative corneal ulcers.

PC2 from 10.2±0,2 16,2±0,4 testifies complicated cases, i.e. points to the 2nd form of the disease.

Give detailed clinical examples for each form of flow GER.

Example 1. Patient B., 59 years old (IB N 011585/925 for 2001) was admitted to the hospital trauma Center and emergency conditions of the organ of vision on the 3rd day after the disease with a diagnosis of Bacterial keratitis of the left eye. History - while walking down the street during a strong wind hit the mote in his left eye. Not treated. Admission: left eye - severe photophobia, lacrimation, excessive mucous discharge on the cornea, paracentral to 5 hours is infiltration in diameter 2×3 mm in the intermediate layers, a smooth edge, in the center of a grayish tinge. The right eye is calm. Visual acuity: right eye and 1.0 in the left eye - 0,005 not corrects. Immediately assigned to conservative treatment. Immunological parameters of the lacrimal fluid, taken on the first day of admission: OK IL-8=1320 PG/ml; OK IL-1β=162 PG/ml; PC=8,15. This value PC testified about the possibility of a favorable course. The prediction was confirmed. After treatment of the eyes calmed down, the bottom of the ulcer cleared occurred epithelialization of the surface, p is s on day 19, however, it was a soft spot in the paracentral region of the cornea. The vision in the left eye of 0.1 s cor - 2,0° 0,5.

Example 2. Patient S., aged 52 (IB N 001534/124 2001) was admitted to the hospital trauma Center and emergency conditions of the organ of vision on the 5th day after the disease with a diagnosis of Bacterial keratitis of the left eye.

From the anamnesis - the cause of the disease is not defined. Admission: left eye is irritated, expressed mixed infection, photophobia, lacrimation, corneal - ulcer in the paracentral region down to a diameter of 3×4 mm, in the middle of the ulcer edge is smooth, the bottom is covered with a coating of grayish-white color. Reflex pink. Visual acuity of the right eye and 1.0 in the left eye - 0,005 not cor. Assigned to conservative treatment. Immunological indexes in the first day of admission: OK 1L-8=2630 PG/ml; CA 1L-1β=172 PG/ml; PC=15,3.

In this case confirmed the forecast of the complicated course of bacterial keratitis. Despite intensive therapy, on the 8th day there was a sudden activation of infection and purulent fusion of the cornea, and therefore in the emergency order was made operative treatment - penetrating keratoplasty.

The claimed method examined 38 patients, with 3 of them immunologic parameters tears were not consistent with the form of the expected course. This amounted to 7.9%. Therefore, the prediction of the shape of the flow ulcers is agovice on the basis of indicators of the concentration of IL-8 and IL-1β tears in the disease onset can be determined with an accuracy of 92.1%.

Thus, the use of the proposed method of predicting the course of bacterial keratitis allows already in the first days of the disease to predict the form of the disease to assign differentiated, pathogenetically substantiated treatment and timely execution of operations keratoplasty. This method can be applied in any hospital where there is an immunological laboratory, so it is available, and therefore, “actionable”.

A method for predicting the clinical course of bacterial keratitis by examining the tear fluid, characterized in that determine the content of interleukin 8 (IL-8) and the content of interleukin 1 beta (IL-1β) and I hope prognostic factor (PC) by dividing the first figure by the second formula

and largest PC below 10,0 predicts favorable over and above 10.0 - unfavorable course of the disease.

 

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