RU2740362C1 - Diagnostic technique for severity of chronic pharyngitis - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to otorhinolaryngology, and concerns a diagnostic technique for severity of chronic pharyngitis. Substance of the method consists in the fact that chromosome-mass spectrometry of the smear from the mouth is carried out, the amount of microbial cells Lactobacillus spp is determined. and Bifidobacterium spp. If observing Lactobacillus spp. within 0-5211*10⁵ cells/gram of sample, and Bifidobacterium spp. within 71-1259*10⁵ cells/gram of the sample, the average severity of the disease is diagnosed. If observing Lactobacillus spp. within 5212-6180*10⁵ cells/gram, and Bifidobacterium spp. within range of 0-70*10⁵ or 1300-4462*10⁵ cells/gram of the sample, the severe course of the disease is diagnosed.

EFFECT: use of the method enables high-accuracy diagnosis of the severity of chronic pharyngitis.

1 cl, 10 ex

Description

The invention relates to medicine, namely to otorhinolaryngology, microbiology, immunology, therapy and can be used to diagnose the course of chronic pharyngitis.

Chronic pharyngitis (CP) with frequent exacerbations is a socially significant problem: 1% of the working-age population of the Russian Federation, more than 70% of patients visiting otorhinolaryngologists suffer from CP.

Complaints of pain, sore throat, sensation of mucus, "lump", paroxysmal cough, sleep disturbance have been troubling patients for many years. Treatment by otorhinolaryngologists, immunologists, general practitioners, therapists has a temporary effect, and therefore the quality of life in people with CP is significantly reduced.

According to the literature, the main etiological factors in CP are Streptococcus groups A, C and G, Staphylococcus spp. And Staphyl. aureus, Streptococcus pneumoniae, Chlamydia trachomatis, Moraxella catarrhalis, Candida spp., gram-negative bacteria, etc. (Cohen JF, Cohen R., Levy C. Selective testing strategies for diagnosing group A streptococcal infection in children with pharyngitis: a systematic review and prospective multicenter external validation study, CMAJ, 2015, Jan 6.187 (1), P. 23-32).

It is known that the course of a chronic inflammatory process on the posterior pharyngeal wall depends on the composition, virulence, and the amount of microflora; the state of local immunity; features of the mucous membrane: its innervation, blood circulation, degree of moisture; the presence of concomitant diseases.

The identification of severity criteria (laboratory parameters) in CP of recurrent course is of great clinical importance - it allows the development of more effective methods of treatment that reduce the frequency of exacerbations of CP.

In modern domestic and foreign literature, there are no statistically reliable methods for diagnosing the course of pharyngitis with frequent exacerbations. This is due to the fact that the pathogenetic basis of frequent exacerbation of CP is still not fully understood.

A known method for assessing the severity of the course of pharyngitis (Atnasheva PP "Indicators of the immune system in patients with chronic decompensated tonsillitis in combination with various forms of chronic pharyngitis", author's thesis for the competition for academic step, Ph.D., St. Petersburg, 2005, p. 4), including the use of immunological methods - detection of the pharyngeal antigen and antibodies to it, as a criterion for assessing the degree of destruction of the posterior pharyngeal wall in patients with chronic decompensated tonsillitis in combination with various forms of pharyngitis.

The disadvantage of this method is the low accuracy of the method for determining the severity of the course of chronic pharyngitis, since it is simultaneously considered in combination with a concomitant disease.

A known method for diagnosing the course of streptococcal pharyngitis in children (Attia MW, Zaoutis T, Klein JD, Meier FA. Performance of a Predictive Model for Streptococcal Pharyngitis in Children. Arch Pediatr Adolesc Med. 2001; 155 (6): 687-691. Doi: 10.1001 / archpedi.155.6.687), including the identification of a number of clinical and laboratory signs: swelling of the tonsils, pain and enlargement of the cervical lymph nodes, as well as the absence of rhinorrhea. Serotyping was performed using the PATHoDx latex agglutination kit (Remel Inc). A second swab was used for rapid antigen detection using the TestPackPlus (Abbott Laboratories, Abbott Park, Ill). Patients were evaluated for GABHS-positive throat culture. The obtained complex clinical and laboratory information was used to diagnose the course of the disease. According to the results of laboratory examination, the sensitivity, specificity, positive and negative diagnostic indicators were determined.

The disadvantage of this method is that it can be used to assess the prognosis of the course of pharyngitis caused only by "streptococcal microorganisms", the method is laborious.

The closest method, chosen as a prototype, is a method for diagnosing the severity of the course of diseases, including conducting chromatography-mass spectrometry (MSMM) with the determination of microorganisms and recalculating the level of these microorganisms for the number of microbial cells, analysis of the results obtained (V.V. Pchelyakova. Personalized approach to diagnostics, treatment and prevention of diseases, https://vspu2019.ipu.ru/files/vspu/Reviews%20VSPU-2019/18%20U_A1/PchelyakovaVV.pdf). At the same time, the concentrations of 57 microorganisms, their total content, the levels of endotoxin and plasmalogen (a useful microbial metabolite) are determined from different biological media; further, the values of the indicators in the examined person are compared with the norms (obtained in the group of healthy persons): if the indicator increases by more than 2 times, the result is assessed as “deviation from the norm”.

The disadvantages of this method are the large volume of analyzed indicators, which complicates the method and requires additional time, requires special knowledge.

The objective of the present invention is to eliminate these disadvantages, high accuracy of the diagnostics of the severity of pharyngitis, reduce the time to obtain results, by taking into account the quantitative data of more informative markers.

To solve this problem, it was proposed to use the MSMM method, based on the direct analysis of biological fluid for the content of marker substances of microorganisms. This can significantly reduce the diagnostic time, because the MSMM method excludes the stage of sowing colonies, which takes at least a day. Determination of microorganisms and recalculation of the level of these microorganisms into the number of microbial cells, analysis of the results are carried out. What is new is that the number of microbial cells of Lactobacillus spp. Is determined as markers of the severity of recurrent CP. and Bifidobacterium spp. in a smear from the throat and: with the value of Lactobacillus spp. within 0-5211 * 10 ⁵ cells / gram of sample, and Bifidobacterium spp. within 71-1259 * 10 ⁵ cells / gram of the sample, the average severity of the disease is diagnosed; with the value of Lactobacillus spp. within 5212-6180 * 10 ⁵ cells / gram, and Bifidobacterium spp. within the range of 0-70 * 10 ⁵ or 1300-4462 * 10 ⁵ cells / gram of the sample, a severe course of the disease is diagnosed.

The invention is of great practical importance, since laboratory severity criteria are a tool for diagnosing the course of the disease, they are important for the development of more effective treatment.

In comparison with traditional methods of bacteriological research, the use of gas chromatography in combination with mass spectrometry can significantly save research time.

The method is carried out as follows.

In a patient with chronic pharyngitis, a smear is taken from the throat, which is examined using the MSMM method on a gas chromatograph. A sample of body fluids (throat swab) is processed immediately after collection. If immediate analysis is impossible, it is preserved in organic solvents or frozen at -5 ° C.

The previously formed data bank on the chemical composition of clinically important microorganisms serves as the basis for determining the genus of microorganisms. Substances or their combinations specific to specific genera or species were selected from them. Since the detection is carried out against the background of a biological fluid, it is required to take into account the monomeric components of complex lipids of the fluid itself in the insoluble sediment. Thus, for the express analysis of microorganism markers, the following are required: a gas chromatography-mass spectrometer, a calculation program and a database (Platonova A.G., Osipov G.A., Boyko N.B., Kirillova N.V., Rodionov G G. Chromato-mass spectrometric study of microbial fatty acids in human biological fluids and their clinical significance, Clinical laboratory diagnostics. 2015. No. 12. URL: https://cyberleninka.ru/article/n/hromato-mass-spektrometricheskoe- issledovanie-mikrobnyh-zhirnyh-kislot-v-biologicheskih-zhidkostyah-cheloveka-i-ih-klinicheskaya).

Studies have shown that the most informative markers for determining the severity of chronic pharyngitis are Bifidobacterium spp. and Lactobacillus, therefore, when conducting gas chromatography-mass spectrometry, their presence and quantity are revealed. The level of these microorganisms is recalculated for the number of microbial cells 10 ⁵ cells / gram. The concentration levels of these microbes indicate the severity of pharyngitis - moderate or severe: with a value of Lactobacillus spp. In the range of 0-5211 * 10 ⁵ cells / gram of the sample, and Bifidobacterium spp. In the range of 71-1259 * 10 ⁵ cells / gram of the sample, average the severity of the disease; with the value of Lactobacillus spp. within 5212-6180 * 10 ⁵ cells / gram, and Bifidobacterium spp. within the range of 0-70 * 10 ⁵ or 1300-4462 * 10 ⁵ cells / gram of the sample, a severe course of the disease is diagnosed.

Example 1.

Patient I., 45 years old, diagnosed with chronic pharyngitis, exacerbation. The diagnosis was established 5 years ago. The patient was examined according to the proposed method. Throat swab chromato-mass spectrometry (MSMM) showed Bifidobacterium spp. 71 * 10 ⁵ cells / gram, Lactobacillus 1211 * 10 ⁵ cells / gram, which allows diagnosing the average severity of CP.

Was carried out standard treatment (local - antiseptics, drugs - probiotics, to restore the initially reduced level of bifidobacteria). After 2 months - with repeated examination of MSMM - the content of Bifidobacterium spp. 260 * 10 ⁵ cells / gram, Lactobacillus 1550 * 10 ⁵ cells / gram, diagnosed with a moderate severity of the disease.

For the prevention of exacerbations of CP 2 times a year (in autumn and winter) the patient was recommended to carry out repeated courses of treatment with the use of drugs - probiotics.

When observing the patient for a year, no recurrences of CP were noted.

Example 2.

Patient G., 38 years old, diagnosed with CP, established a year ago (after removal of the tonsils).

A smear from the posterior pharyngeal wall was examined - the content of Bifidobacterium spp. 1259 * 10 ⁵ cells / gram, Lactobacillus 2450 * 10 ⁵ cells / gram, which made it possible to diagnose the moderate severity of CP.

The basic therapy (sanation of the nasopharynx) was carried out, after 2 months with a second examination, the level of Bifidobacterium spp. 950 * 10 ⁵ cells / gram, Lactobacillus 1800 * 10 ⁵ cells / gram. Thus, despite the therapy, the course of the disease remained as moderate.

For the prevention of exacerbations, the patient is recommended 2 times a year (autumn, winter), to examine MSMM from a throat swab, in case of a violation of the composition and level of microbiota - to carry out preventive treatment using local antiseptics.

Subsequently, the patient was followed up for a year - no recurrence of CP was noted.

Example 3.

Patient P., M., 43 years old, diagnosed with CP, established 10 years ago.

Throat smear study was carried out - MSMM: the content of Bifidobacterium spp. 71 * 10 ⁵ cells / gram, Lactobacillus 0, which made it possible to diagnose the moderate severity of CP.

2 months after the treatment (local antiseptics, a complex probiotic preparation), the patient had an exacerbation of CP, repeated examination of MSMM: the content of Bifidobacterium spp. 0 cells / gram, Lactobacillus 5212 * 10 ⁵ cells / gram, which indicates a severe course of CP.

A repeated course of treatment with the use of a probiotic preparation containing bifidobacteria, the level of which in the patient is significantly reduced (for the prevention of relapses - every 3 months) is recommended.

The patient was observed for a year - exacerbations of CP were noted only once.

Example 4

Patient R., 41 years old, diagnosed with CP, established 3 years ago, an examination was carried out - MSMM smear from the posterior pharyngeal wall, revealed: Bifidobacterium spp. 71 * 10 ⁵ cells / gram, Lactobacillus 5211 * 10 ⁵ cells / gram - moderate.

2 months after the complex therapy - local antiseptics and immunotherapy with bacterial lysates (imudon), the content of Bifidobacterium spp. 350 * 10 ⁵ cells / gram, Lactobacillus 4530 * 10 ⁵ cells / gram, which indicates a severe course of CP. It was concluded that the prescribed therapy was ineffective, after its change, repeated courses of treatment were recommended to prevent exacerbations of CP, which were carried out 3 times a year (spring, autumn, winter).

The patient was monitored for a year - there were no relapses of CP.

Example 5

Patient T., m., 47 years old, diagnosed with CP, established 10 years ago, examination of MSMM from the throat was carried out, revealed: Bifidobacterium spp. 0, Lactobacillus 5220 * 10 ⁵ cells / gram, which makes it possible to diagnose a severe course of CP, standard therapy with the inclusion of a probiotic containing bifidobacteria was prescribed.

After 2 months, with a second examination - MSMM from the throat: Bifidobacterium spp. 73 * 10 ⁵ cells / gram, Lactobacillus 5210 * 10 ⁵ cells / gram. These values correspond to the moderate severity of CP.

It is recommended to conduct preventive courses of therapy (to prevent exacerbations of CP) 2 times a year (autumn, winter). The patient was observed during the year - no exacerbations of CP were noted.

Example 6

Patient P., 53 years old, diagnosed with CP, established 6 years ago, a study of the state of the microbiota of a smear from the throat by chromatography-mass spectrometry was carried out, revealed: Bifidobacterium spp. 70 * 10 ⁵ cells / gram, Lactobacillus 6000 * 10 ⁵ cells / gram, which indicates a severe course of CP.

The standard treatment of CP with the inclusion of a probiotic preparation containing bifidobacteria was carried out.

2 months after treatment, with repeated examination of MSMM - throat swab: Bifidobacterium spp. 350 * 10 ⁵ cells / gram, Lactobacillus 4200 * 10 ⁵ cells / gram, this indicates a moderate severity of CP.

It is recommended to carry out preventive treatment courses - 3 times a year (spring, autumn, winter). The patient was monitored for a year - exacerbations of CP were noted only once in winter, as concomitant with influenza.

Example 7.

Patient Z., 63 years old, diagnosed with CP, established 20 years ago, an examination of MSMM from a throat swab was carried out: the content of Bifidobacterium spp. 1800 * 10 ⁵ cells / gram, Lactobacillus 5500 * 10 ⁵ cells / gram. This makes it possible to diagnose severe CP.

Conducted standard CP therapy, 2 months after treatment - MSMM: the content of Bifidobacterium spp. 1300 * 10 ⁵ cells / gram, Lactobacillus 5212 * 10 ⁵ cells / gram, which revealed the persistence of a severe course of CP, with exacerbations after 1-2 months. It is recommended to change the drugs used and to conduct basic therapy courses regularly in spring, autumn and winter to prevent exacerbations of CP.

The patient was followed up for a year - no recurrences of CP were noted.

Example 8.

Patient X., 41 years old, diagnosed with CP, established 5 years ago, when examining a smear from the posterior pharyngeal wall using the MSMM method, it was revealed: Bifidobacterium spp. 4462 * 10 ⁵ cells / gram, Lactobacillus 5600 * 10 ⁵ cells / gram, which indicates a severe course of CP.

A course of standard antiseptic therapy was carried out. 3 months after treatment, repeated examination - MSMM revealed: Bifidobacterium spp. 1250 * 10 ⁵ cells / gram, Lactobacillus 3400 * 10 ⁵ cells / gram, which indicates an average severity of CP, i.e. in this patient, the course of CP responds positively to treatment.

It is recommended to carry out preventive therapy courses in autumn and spring. The patient was monitored for a year, no relapses were noted.

Example 9.

Patient D., 39 years old, the diagnosis - HF, 14 years old (installed after the removal of the tonsils), throat swab method MSMM identified: Bifidobacterium spp.0, Lactobacillus 5212 * 10 ⁵ cells / gram, which allows you to diagnose severity of the CP.

After a course of standard therapy with the inclusion of a probiotic preparation containing bifidobacteria.

After 2 months, repeated examination of MSMM was carried out, it was found: Bifidobacterium spp. 70 * 10 ⁵ cells / gram, Lactobacillus 5700 * 10 ⁵ cells / gram, which indicates a positive dynamics in the level of Bifidobacterium spp., But the severity, determined by the level of significant laboratory markers, is still assessed as "severe course of CP" ... Therefore, the patient is recommended to undergo repeated courses of therapy to prevent exacerbations of CP, with the inclusion of probiotic preparations (containing bifidobacteria) - 1 time in 3 months.

The patient was observed for 12 months, no exacerbations of CP were noted.

Example 10.

Patient K., 38 years old, diagnosed with CP, established 5 years ago, the patient was examined according to the proposed method - gas chromatography-mass spectrometry to assess the microbiota of the posterior pharyngeal wall, found: Bifidobacterium spp. 4462 * 10 ⁵ cells / gram, Lactobacillus 6180 * 10 ⁵ cells / gram, which indicates a severe course of CP.

One month after treatment, repeated MSMM results showed the following values: Bifidobacterium spp. 1200 * 10 ⁵ cells / gram, Lactobacillus 5100 * 10 ⁵ cells / gram, which indicates an average severity of CP.

It is recommended to carry out repeated courses of treatment 2 times a year (in autumn and winter) to prevent recurrence of CP. During the subsequent observation of the patient for 6 months, there were no exacerbations of CP.

The proposed method for determining the severity of chronic pharyngitis with frequent exacerbations (recurrent course) was used in 96 patients. Observation of the patients was carried out for 3,6,12 months; in 97.9% of cases (94 people), the correctness of establishing the severity of chronic pharyngitis was confirmed: in 67 patients, moderate severity (69.8%), 29 people (30.2%) - severe CP.

Thus, the markers used make it possible to diagnose the severity of the disease and to carry out effective therapy - in patients with severe CP, the number of relapses decreases, the severity of the course changes to medium. If the treatment is ineffective, a treatment is selected that allows the patient to lead to a stable remission of the disease (6-12 months).

The main advantages of the proposed method for determining the severity of a chronic course with frequent exacerbations is a high degree of information content and reliability of diagnostic criteria, which allow, taking into account the obtained values, to correct the course and effectiveness of treatment, thereby improving the quality of life of this category of patients.

Claims (1)

A method for diagnosing the severity of chronic pharyngitis, including a chromatography-mass spectrometric study with the determination of microorganisms and recalculation of the level of these microorganisms for the number of microbial cells, analysis of the results obtained, characterized in that the number of microbial cells of Lactobacillus spp. Is determined as markers of the severity of the course of recurrent CP. and Bifidobacterium spp. in a smear from the throat and with the value of Lactobacillus spp. within 0-5211 * 10 ⁵ cells / gram of sample, and Bifidobacterium spp. within 71-1259 * 10 ⁵ cells / gram of the sample, the average severity of the disease is diagnosed; with the value of Lactobacillus spp. within 5212-6180 * 10 ⁵ cells / gram, and Bifidobacterium spp. within the range of 0-70 * 10 ⁵ or 1300-4462 * 10 ⁵ cells / gram of the sample, a severe course of the disease is diagnosed.