Description

Method of determination Immunoassay.

Material under study Serum

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Mycoplasmas - a group of intracellular microorganisms - gram-negative bacteria 115 - 200 nm in size, which do not have a dense cell wall, covered with a three-layer cytoplasmic membrane. Several strains of mycoplasmas have been described.

Conventionally, mycoplasmas are divided into 6 groups, depending on the diseases they cause in humans. The group of mycoplasmas that cause lesions of the urogenital tract in men and women includes Mycoplasma hominis type I and type II, Ureaplasma urealyticum.

Mycoplasmas are characterized by polymorphism and a peculiar life cycle. The source of infection is a person with mycoplasmosis, or a healthy carrier of mycoplasmas.

Mycoplasma infections of the urogenital tract occupy one of the leading places among STIs. They are often combined with gonococci, Trichomonas and opportunistic microorganisms, are transmitted through sexual contact, can cause non-gonococcal urethritis and prostatitis, inflammatory diseases of the small pelvis, pathology of pregnancy and fetus, infertility in women and men, as well as perinatal infection of newborns.

Diagnosis of Mycoplasma hominis infection using microbiological methods is difficult because it is difficult to cultivate this microorganism in vitro. Adequate modern method diagnosis of M. hominis infection is a PCR method aimed at identifying the DNA of the pathogen (in the INVITRO laboratory tests,).

Serological methods (detection of antibodies in blood serum) are of lesser use, since, due to the intracellular localization of M. hominis, the body's immune response against these microorganisms is often weakly expressed. A positive test result for IgM antibodies may indicate the likelihood of an ongoing infection.

Mycoplasmatosis: causes, symptoms and diagnosis of the disease

Among a sufficiently large number of mycoplasmas found in humans, only 4 species can cause disease under certain conditions. One of them - Mycoplasma pneumonia - affects the respiratory system, causing inflammatory diseases of the throat, bronchi, and lungs. The other three - Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealiticum - are the causative agents of urogenital mycoplasmosis - one of the most common sexually transmitted diseases.

Mycoplasma causes:

1. Diseases of the upper respiratory tract(atypical pneumonia, mycoplasmal bronchitis, etc.).

The causative agent of this group of diseases is a microorganism of the species Mycoplasma pneumoniae.

The main route of transmission of diseases: - airborne.

The source of infection is a sick person and a healthy one (carrier).

The possibility of infection persists throughout the year, but in the autumn-winter period, infection is activated.

Clinical manifestations:

cough is the most common symptom of respiratory tract infection in patients with mycoplasma infection. As a rule, a dry, hysterical cough with light sputum is present throughout the disease, but among those who cough only 3-10% of patients with pneumonia.

Diagnostics:

one of their main assets laboratory diagnostics infections caused by Mycoplasma pneumoniae are serological studies, partly due to their wide availability and ease of sampling - used deoxygenated blood to detect antibodies (Ig A, IgM, IgG).

These methods are widely used in clinical practice. In recent years, an increase in their sensitivity has been achieved through the separate detection of different classes of antibodies (IgM and IgA). Enhanced Level IgM is a reliable indicator of mycoplasmal infection in children. In adults, methods based on the determination of IgA have a higher sensitivity.

Ig G is an indicator of a current or past Mycoplasma pneumoniae infection, these antibodies appear later than Ig A and Ig M, and persist for a longer time (more than a year).

Another modern method for diagnosing Mycoplasma pneumoniae is PCR diagnostics. PCR (polymerase chain reaction) is a method that allows you to find in the studied clinical material a small section of genetic information (DNA) of any organism among a huge number of other sections and multiply it many times.

The clinical material for the study can be venous blood, saliva, sputum, separated from the ear, throat and nose.

2. Diseases of the urogenital tract in men and women (Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealiticum).

Currently, mycoplasmas are considered to be opportunistic microbes. Only Mycoplasma genitalium is considered by most researchers as a pathogenic microorganism that can cause urethritis, epidymitis in men, and cervicitis, vaginitis, inflammatory diseases of the pelvic organs and pregnancy pathology in women.

The frequency of detection of Mycoplasma hominis, Ureaplasma urealiticum varies widely and ranges from 10% to 50%. These microorganisms are often detected in clinically healthy individuals and, being opportunistic microorganisms, can normally colonize the organs of the urogenital system.

Genital mycoplasmas (Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma urealiticum) can be infected in several ways:

during sexual contact;

when the infection is transmitted from mother to fetus through an infected placenta or during childbirth;

in transplantation (transplantation) of organs;

indirectly (in women, especially girls, through household items).

Clinical manifestations of urethritis caused by genital mycoplasmas:

dysuria (itching, burning, pain when urinating);

discomfort, itching, burning in the urethra;

frequent urination or urge to urinate;

pain during sexual intercourse (dyspanuria).

Clinical manifestations of vaginitis caused by genital mycoplasmas:

mucous or mucopurulent discharge from the genital tract;

discomfort, itching, burning on the mucous membrane of the genital tract.

Clinical manifestations of cervicitis caused by genital mycoplasmas:

spotting after sexual intercourse;

discomfort or pain in the lower abdomen;

soreness during sexual intercourse.

Diagnosis of urogenital infectious diseases

Indications for examination for Mycoplasma hominis, Ureaplasma urealiticum:

clinical and / or laboratory signs of inflammation of the urogenital tract (urethritis, prostatitis, cystitis, cervicitis, cervical erosion, pyelonephritis, vaginitis);

recurrent pathological processes associated with an imbalance in the vaginal flora (bacterial vaginosis);

pre-gravid (pregnancy planning) examination of sexual partners;

upcoming surgical manipulations on the pelvic organs with a high risk of infectious complications;

the presence of a burdened obstetric or gynecological history (miscarriage, perinatal losses, infertility);

the possibility of infection of the fetus with a complicated course.

An additional indication for testing for the presence of Mycoplasma genitalium is the detection of Mycoplasma genitalium in one of the partners, as well as a change of sexual partner in the absence of the use of barrier methods of contraception.

Material for laboratory research for the presence of urogenital infections are obtained: 1) in men - from the urethra, prostate gland, and it is also possible to conduct a study of the ejaculate and the first portion of morning urine, 2) in women - from the urethra, vagina and cervical canal (cervix).

For the qualitative laboratory diagnosis of urogenital infections, it is important to correctly obtain clinical material for research from the patient. To obtain the most reliable test result, it is recommended to comply with a number of requirements:

Donate biomaterial before the start of treatment or not earlier than 1 month after the end of antibiotic therapy;

Observe the time frame for obtaining biomaterial: a) from the urethra not earlier than 3 hours after the last urination, b) in the presence of abundant urethral discharge - 15-20 minutes after urination, c) from the cervical canal and vagina before menstruation or after 1-2 days after its completion;

To carry out the taking of biomaterial in sufficient quantities for laboratory research.

Methods of laboratory diagnostics of urogenital infectious diseases

Currently, for the purpose of diagnosing urogenital infections, a number of methods are used that differ in sensitivity, specificity, ease of use and general availability.

The independent laboratory INVITRO offers a wide range of tests to detect the presence of mycoplasma infection.

For the identification of Mycoplasma genitalium, the only research method is the PCR method. PCR diagnostics of Mycoplasma hominis is based on the detection of the genetic material of the pathogen (DNA) in the biological material.

The advantages of the method are:

the possibility of using a variety of biological material (scraping, urine, prostate secretion, semen, saliva, synovial fluid) depending on the location of the alleged pathogen localization;

high sensitivity of the method allows early diagnosis of urogenital infections;

high speed of analysis.

To detect Mycoplasma hominis and Ureaplasma urealiticum, a cultural (bacteriological) study is carried out with a quantitative determination of isolated microorganisms and sensitivity to antibiotics. Clinically significant is the detection of Mycoplasma hominis and Ureaplasma urealiticum in an amount of more than 10^4 cfu/ml.

In addition, in order to assess the state of the epithelium of the genital organs, the presence of an inflammatory process and concomitant sexually transmitted infections, it is recommended to conduct a microscopic examination of a Gram-stained smear.

Literature

1. Manual of Infectious Diseases with an Atlas of Infectious Pathology. Edited by Yu.V. Lobzina, S.S. Kozlova, A.N. Uskov. www.infectology.spb.ru, St. Petersburg. 2000

Indications for appointment

Positive result:

1. likely current infection with Mycoplasma hominis;
2. bacillus carrying.

Negative result:

1. early or late terms of Mycoplasma hominis infection;
2. weak immune response to Mycoplasma hominis;
3. no infection (with negative PCR results).

* The positivity ratio (PC) is the ratio of the optical density of the patient sample to the threshold value. KP - the coefficient of positivity is a universal indicator used in high-quality enzyme immunoassays. The CP characterizes the degree of positivity of the test sample and may be useful to the doctor for the correct interpretation of the result. Since the positivity coefficient does not correlate linearly with the concentration of antibodies in the sample, it is not recommended to use the CP for dynamic monitoring of patients, including monitoring the effectiveness of treatment.

Mycoplasma pneumoniae is one of the major respiratory bacterial pathogens, disease-causing respiratory tract. The sooner the diagnosis is made and the pathogen is established, the more effective the treatment will be and the less severe the consequences. For research, secretions from the mucous membranes of the respiratory tract or blood are taken from infected patients. These can be sputum samplings, scrapings from the surface of the walls of the larynx, or bronchoalveolar lavage. The material is taken depending on the alleged foci of the spread of the disease in the patient. At a temperature of 2 to 8 degrees, the material is suitable for research throughout the day. At a temperature of about -20 degrees, the collected material is stored for up to a week, retaining its properties. A pediatrician prescribes an analysis for children, a therapist for adults. Ingoda, in severe cases of pneumonia, a pulmonologist or an infectious disease specialist is connected to the treatment.

The most effective, although quite expensive, is the PCR method for detecting the tiny mycoplasma pneumoniae virus in sputum or other material. The polymerase chain reaction is a fairly effective method by which it is possible to detect a separate section of mycoplasma DNA in the material that is submitted to the laboratory for research and separate it from other sections located in this material. This material is then laboratory multiplied and examined. PCR studies are based on DNA replication. Doctors consider this study one of the most reliable in order to timely identify the virus in the analyzes - Mycoplasma pneumonia. The sensitivity of this method is about 93%, and its specificity reaches 98%.

Second, but no less proven and effective method study of the presence in the body of mycoplasma pneumonia, is the analysis of ELISA. Only enzyme immunoassay is able to detect serological markers of IgG and IgM, which are formed to mycoplasma. In this study, the patient's blood is provided as a material to the laboratory, then the blood serum is separated to detect class M or G antibodies in it.

The results of the analyzes depend on the presence of the virus in the test material. Mycoplasma pneumonia in the presence of IgG antibodies will be positive if the number in titers is more than 20 IU/L. This indicates a current infection or a recent illness with a residual presence of antibodies in the material. A group of IgG antibodies to mycoplasma is formed approximately two weeks after the infection has entered the body. They can be found in the blood for 2 years even after the disease is completely cured.

A negative analysis is considered when the amount of IgG antibodies is less than 16 IU/l. With such titers, the disease is absent or the analysis was taken at a very early term when antibodies to mycoplasma pneumonia IgG have not yet begun to form. For the final determination of the diagnosis, a repeated examination of the material is carried out. If after two weeks the patient's condition does not improve and the titers increase, the progression of the disease should be considered.

The presence of IgM antibodies to mycoplasma indicates early detection of the disease. In the later stages, these specific immunoglobulins may no longer be visible in the analyzes, but this does not mean the absence of mycoplasma. Doctors usually look at the total value of these quantities.

Antibodies of the LgG group in mycoplasma pneumonia indicate the presence of chlamydia in a patient.

Each patient must be notified that he will have to take a PCR or ELISA test to determine the diagnosis. Although these studies do not require any special preparation, you should still know that:

• For the analysis of ELISA for the determination of antibodies of the groups LgG, IgG, IgM, only venous blood is taken.
• Patients are not allowed to smoke before donating blood (30 minutes).
• Patients are required to inform their doctor if they have an autoimmune disease.

Researchers suggest that three types of tiny bacteria are responsible for a number of pathologies of the respiratory system, urogenital tract, and digestive system. These are unicellular microorganisms Mycoplasma pneumoniae, M. genitalium, M. hominis, which do not have a strong cell membrane. Mycoplasmas often infect the epithelial cells of the upper respiratory tract. In second place are infectious diseases of the genitourinary system. Active reproduction of bacteria disrupts the functions of many organs.

Mycoplasma pneumoniae causes tonsillopharyngitis, sinusitis, tracheobronchitis, mild atypical pneumonia. The child feels a sore throat, he has an obsessive cough, subfebrile temperature. Symptoms and treatment of mycoplasma in children is similar to SARS; cases of occurrence of mixed infections are known. Further reproduction of pathogens in the respiratory tract often leads to the development of pneumonia.

Mycoplasmas are found in association with ureaplasma, chlamydia, combined with viral infection, namely with adenoviruses, influenza and parainfluenza viruses.

Outbreaks of acute respiratory diseases in children from 5 to 15 years old are recorded throughout the cold period of the year. In the structure of acute respiratory infections, mycoplasmosis accounts for only about 5%, but this figure increases by about 10 times every 2–4 years during epidemics. Mycoplasma causes up to 20% of acute pneumonia.

Symptoms and diagnosis of mycoplasmosis of the upper respiratory tract

The incubation period of the pathogen ranges from 3–10 days to 4 weeks. The difficulty of recognizing the respiratory form of mycoplasma lies in the fact that clinical picture usually resembles SARS. Children, unlike adults, react more sharply to the activity of the pathogen. There are manifestations of intoxication, runny nose, paroxysmal cough, which can result in vomiting.

Initial symptoms of mycoplasma in a child:

1. Elevated temperature persists for 5–10 days up to 37.5°C;
2. perspiration, itching and sore throat;
3. runny nose, stuffy nose;
4. conjunctivitis;
5. headache;
6. dry cough;
7. weakness.

When examining the throat, redness of the oropharyngeal mucosa can be noticed. It is the similarity of the course of respiratory mycoplasmosis in children with ARVI that makes it difficult to diagnose the disease. Parents give the child antitussives, syrups to improve expectoration. However, such treatment most often does not work, and the cough continues for several months. Against the background of mycoplasma activity in the upper respiratory tract, newborns, premature infants and children under 8 years of age develop sinusitis, bronchitis, and pneumonia.

Mycoplasmosis of the lungs

Clinical manifestations of mycoplasmal pneumonia resemble chlamydia of the lungs. Therapy of diseases also has many common features. The similarity of the two different microbial infections is due to their small size compared to other bacteria and the lack of a solid cell wall. Mycoplasmas cannot be seen under a conventional light microscope.

Signs of the pulmonary form of mycoplasmosis in children:

• the disease begins suddenly or as a continuation of SARS;
• chills, fever up to 39 ° C;
• dry cough is replaced by wet;
• sputum scanty, purulent;
• headaches and muscle pain.

The pediatrician, listening to the child's lungs, notes hard breathing and dry wheezing. X-ray shows that there are scattered foci of inflammation in the tissues of the lungs. The doctor suggests taking an analysis for mycoplasma in children - a blood test from a vein that will confirm or refute the initial diagnosis. For the recognition of mycoplasma infection, methods of enzyme immunoassay and polymerase chain reaction(respectively, ELISA and PCR). The accumulation of antibodies belonging to the IgG and IgM types occurs during the body's immune response to the activity of mycoplasma.

Mycoplasmosis of the kidneys and other organs

Children can become infected from adults through direct contact - this is sleeping in a shared bed, using one toilet seat, towels. It happens that the kindergarten staff becomes the source of mycoplasma. In the respiratory and urogenital form of mycoplasmosis, epithelial cells are mainly affected. Dystrophic changes in the tissue begin, its necrosis.

Infection of the urogenital system in adolescents leads to cystitis, pyelonephritis, vaginitis. Mycoplasmas initiate pathological processes in the liver, in the small intestine, in various departments brain and spinal cord. Mycoplasmosis in girls adolescence manifests itself in the form of vulvovaginitis and mild lesions of the urogenital tract. The course of the disease is most often asymptomatic, in the case of severe forms pain occurs in the lower abdomen, mucous discharge appears.

Mycoplasma in the blood of a child can cause the development of a generalized form, which is characterized by damage respiratory system and a number internal organs. The liver increases in size, jaundice begins. Perhaps the development of meningitis, brain abscess, meningoencephalitis. A pink rash appears on the body, watery and red eyes (conjunctivitis).

Treatment of a bacterial infection

If only a runny nose worries, the temperature is subfebrile, then antibacterial drugs will not be required. Antibiotic treatment is a specific therapy for mycoplasmosis. The drugs of choice are macrolides, fluoroquinolones, tetracyclines. Other medicines are given depending on the symptoms.

Oral antibiotics:

1. Erythromycin - 20–50 mg per 1 kg of body weight per day for 5–7 days. daily dose divided into three doses.
2. Clarithromycin n - 15 mg per 1 kg of body weight. Give in the morning and evening, with an interval between doses of 12 hours.
3. Azithromycin - 10 mg per 1 kg of body weight on the first day. In the next 3-4 days - 5-10 mg per kg of body weight per day.
4. Clindamycin - 20 mg per 1 kg of weight per day 2 times a day.

Mycoplasmas grow more slowly than other bacteria. Therefore, the duration of treatment is not 5–12 days, but 2–3 weeks.

Clindamycin belongs to the lincosamide antibiotics. Clarithromycin, erythromycin and azithromycin belong to the group of macrolides. Tetracycline antibiotics are being used less and less due to the spread of bacterial strains resistant to them. There is a practice of combining antimicrobial drugs that differ in the mechanism of action. For example, doctors may prescribe a combination of erythromycin and tetracycline. Another option is to change the antibiotic during a long course of treatment. The choice of remedy is influenced by an allergy in a child to substances belonging to certain groups of antibacterial drugs.

Tablet forms of antibiotics are more difficult to give to babies, especially if it is necessary to calculate the dose and divide one capsule into several doses. Doctors recommend treating children under 8–12 years old with suspensions that are prepared from an antibacterial substance in the form of powder and water. They produce such funds in glass vials, supply with a dosing pipette, a convenient measuring cup or spoon. The drug in children's dosage is usually sweet in taste.

Concomitant treatment (by symptoms)

A child infected with mycoplasma is given with high temperature non-steroidal anti-inflammatory drugs to alleviate the patient's condition. Children are prescribed ibuprofen or paracetamol in the form of a suspension for oral administration, rectal suppositories. You can use a vasoconstrictor nasal spray, take antihistamine drops or syrup inside (drugs "Zirtek" or similar "Zodak", "Loratadin", "Fenistil" for younger patients).

Concomitant treatment reduces irritation and sore throat, but does not affect the causative agent.

Cough remedies, for example "Sinekod", it is recommended to give only in the first days. Then the child will be able to rest from bouts of painful coughing. In the future, the doctor prescribes expectorant drugs to thin and facilitate sputum discharge. Justified use for the treatment of mycoplasma pharmaceutical preparations and folk remedies that strengthen the immune system.

Mycoplasmas in children after an acute period of the disease remain in the body, although in small quantities. Full recovery does not occur, immunity to the pathogen is not developed. Against this background, laryngitis, pharyngitis, bronchitis periodically occur. Often respiratory and urogenital mycoplasmosis becomes chronic.

Mycoplasma prevention

It is recommended to isolate a child with mycoplasmosis from other children for 5-7 days with a respiratory form bacterial infection, for 14–21 days - with a pulmonary variety. The same preventive measures are taken as in other acute diseases of the upper respiratory tract - SARS, influenza, tonsillitis. There are no drugs that a child or adult could take to prevent mycoplasma infection.

Mycoplasma - the causative agent of respiratory and other diseases in a child updated: September 21, 2016 by: admin

More about the study

Mycoplasma pneumonias (sometimes called "atypical pneumonias") account for up to 15-20% of all cases of community-acquired pneumonia. Sometimes they can lead to whole epidemics, especially in children. school age and in closed populations, like the military. The source of infection are both patients and carriers. Infection occurs by airborne droplets, incubation period lasts 2-3 weeks. The symptoms of mycoplasma infection vary. In most cases, the disease is mild and is accompanied by cough, runny nose, sore throat, which persist for several weeks. When the infection spreads to the lower respiratory tract, headaches, intoxication, fever, and muscle pain occur. Pneumonia is most severe in young children and people with weakened immune systems, such as those with HIV.

The diagnosis of "mycoplasmal infection" is often difficult, so several research methods are used, in which serological tests play a leading role.

In response to Mycoplasma pneumoniae infection the immune system produces specific immunoglobulins: IgA, IgM and IgG.

The production of class G immunoglobulins to Mycoplasma pneumoniae does not begin immediately after infection, after about 2-4 weeks, but continues for a long period (a year or more).

The presence of class G immunoglobulins to Mycoplasma pneumoniae in the blood indicates an acute or past illness, a chronic inflammatory process, or reinfection.

What is research used for?

• To confirm the current disease (including reinfection) caused by Mycoplasma pneumoniae.
• For differential diagnosis mycoplasma pneumonia and other infectious diseases of the respiratory tract, such as pneumonia caused by streptococci or staphylococci.
• For the diagnosis of mycoplasmal infection in chronic inflammatory diseases respiratory tract.

When is the study scheduled?

• With symptoms of a disease caused by mycoplasma (unproductive cough that can persist for several weeks, fever, sore throat, headaches and muscle pain).
• If you suspect a chronic or persistent form of Mycoplasma pneumoniae infection, manifested by frequent relapses.

Mycoplasmas (Mycoplasma hominis), Ig A class antibodies by ELISA, qualitative analysis

Mycoplasma hominis is one of the types of mycoplasmas, special microorganisms that occupy an intermediate stage between protozoa, viruses and bacteria. It is opportunistic pathogen...

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Study Description

Preparation for the study:

Special training is not required.

Material under study: Taking blood from a vein

In response to infection with mycoplasma, the immune system begins to produce a number of specific immunoglobulins: IgA, IgM and IgG.

Class A immunoglobulins begin to be produced at the very last turn, after the appearance of IgG, and this production continues for a long time: a year or more. In elderly patients, the level of IgA rises more than the level of IgM.

The presence in the blood of class A immunoglobulins to Mycoplasma hominis indicates the presence acute illness, as well as a chronic or recurrent form of infection.

Method

Enzyme immunoassay (ELISA) is a laboratory immunological method for the qualitative or quantitative determination of various compounds, macromolecules, viruses, etc., which is based on a specific antigen-antibody reaction. The resulting complex is detected using the enzyme as a label for signal registration. Due to the undoubted advantages - ease of use, speed, objective automated recording of results, the possibility of studying immunoglobulins of various classes (which plays a role in the early diagnosis of diseases, their prognosis), ELISA is currently one of the main methods of laboratory diagnostics.

Reference values ​​- norm
(Mycoplasma hominis (mycoplasmosis), IgA antibodies, qualitative, blood)

Information regarding the reference values ​​of the indicators, as well as the very composition of the indicators included in the analysis, may differ slightly depending on the laboratory!

Norm:

The study is qualitative, the result is defined as “positive” / “detected” or “negative” / “not found”

Indications

• Symptoms of acute or chronic mycoplasma infection: itching, burning, pain in the urethra during urination, frequent urge to urinate, discharge from the genital tract, discomfort in the lower abdomen, pain during sexual intercourse.
• Suspicion of a chronic or persistent form of mycoplasma infection, which can cause pregnancy pathologies or infertility.

Increasing values ​​(positive result)

Result "positive"/"detected":

• Current acute mycoplasma infection
• Chronic mycoplasma infection
• Asymptomatic carriage of Mycoplasma hominis,
• The presence of IgA in the absence of IgM indicates reinfection with Mycoplasma hominis.
• In newborns - perinatal infection.

Decreasing values ​​(negative result)

Result "negative"/"not found":

• No Mycoplasma hominis infection
• Infection with Mycoplasma hominis occurs, but sufficient time has not elapsed for an immune response to develop.