Mycoplasma PCR method. Ureaplasma and mycoplasma. Mycoplasmosis: blood test to detect pathogen resistance

And mycoplasma are not absolute pathogens, and their detection in tests does not require treatment, but not if you are planning a pregnancy. When planning, everything is very complicated: (Doctors themselves cannot agree on the need to treat these pathogens.

Therefore, the question of the need Treatment of ureaplasma and mycoplasma should be discussed with a trustworthy personal physician.

Our personal opinion is that “treating with tests” is still not correct. And you should not take antibiotics, provided there are no complaints from the woman, a normal smear on the flora and a complete absence of clinical symptoms.

Ureaplasmas and mycoplasmas have no clinical significance in obstetrics and gynecology. These are causative agents of nonspecific urethritis, more often in men. In 30% of cases or more - representatives of the normal microflora of the genital tract. Their detection by PCR is not an indication for their targeted treatment, even if there are symptoms of an inflammatory process - it is necessary to treat more common pathogens, and since they are chlamydia, and the drugs used against them and urea- and mycoplasmas are the same, then the question of treating myco- and ureaplasmosis has been removed. Even if we accept that they exist and are important, they are still treated with the same drugs, therefore there is no point in identifying them.

Is it necessary to take a culture test for mycoplasma and ureaplasma?

Diagnosis of myco- and ureaplasmosis is not necessary. There is no need to take tests for them - neither blood for antibodies, nor culture (especially since only a few laboratories in the capital actually do it, and determining sensitivity to antibiotics is technically unrealistic; in ordinary places they write PCR results as culture), nor PCR.

If for some reason the analysis is done, you do not need to pay attention to its results; they are not a criterion for making a diagnosis, much less prescribing treatment.

Planning pregnancy and pregnancy itself is not an indication for PCR diagnostics in general, and even more so for PCR diagnostics of urea and mycoplasmas. Management in this case does not differ from the management of non-pregnant women - complaints and a smear.

The treatment is not tests, but complaints. If there are no complaints, and a regular flora smear shows a normal number of leukocytes, no further examination or treatment is needed. If an additional examination is nevertheless done, and something is found in the PCR, this is not a criterion for prescribing treatment. In addition to the lack of clinical significance of urea- and mycoplasmas, it is necessary to remember the high frequency of false-positive PCR results. Prescribing this test in the absence of complaints at all, and in the presence of complaints - before or instead of a smear - is incompetence and a scam of money.

If there are complaints, but the smear done in a good laboratory is good, there are no indications for antibiotics, you need to look for other causes of complaints - dysbacteriosis, concomitant diseases, hormonal imbalance, allergies, papillomatosis.

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If there are complaints and signs of an inflammatory process in the genitourinary system, antibiotic therapy is prescribed - either based on the results of additional examinations (PCR and culture with sensitivity determination) - for various pathogens (chlamydia, gonococci, trichomonas, streptococci, E. coli, etc., etc.), but not against urea and mycoplasmas, or “blindly” - against the main causative agents of such diseases (gonococci and chlamydia). An anti-chlamydial drug is prescribed without fail, in any case, regardless of the test results, since it is the most common pathogen, and since it does not have resistance to anti-chlamydial antibiotics (cultures to determine the sensitivity of chlamydia are also profanation). All myco- and ureaplasmas are sensitive to antichlamydial drugs (with the exception of a certain proportion of ureaplasmas that are resistant to doxycycline). Therefore, even if after some time the pathogenicity and clinical role of these microorganisms is proven, adequate treatment of inflammatory diseases without identifying them will still eliminate them along with chlamydia. Therefore, again, there is no point in defining them. Contrary to what they say now in many commercial centers, treatment in this case does not depend on test results, there is one scheme.

This scheme is very simple and inexpensive; a multicomponent list of antibiotics on two sheets against a positive PCR for ureaplasma is incompetence and a scam. Doxycycline is an old drug, but the main causative agents of inflammatory diseases in gynecology have retained sensitivity to it. However, the duration of treatment is not shorter than 10 days. Equivalent in effectiveness against the main pathogens is a single dose of 1 g of sumamed. For those who continue to be afraid of ureaplasma, this is the drug of choice, since those ureaplasma that are genetically insensitive to doxycycline are sensitive to sumamed. Scientific studies have proven the equivalence of a course of treatment to a single dose of 1 g. Fast, simple, cheap.

Malyarskaya M.M. gynecologist

Mycoplasmosis and ureaplasmosis

It is difficult to give an unambiguous answer to the question of the clinical significance of genital mycoplasmas, at least at this point in time. The fact is that research into their etiological role in various pathological conditions of both the female and male urogenital systems began relatively recently.

If there is a clinic for cervicitis and/or urethritis in women or urethritis in men, then at the initial stage economically It is not advisable to test for genital mycoplasmas. Even if gonococci and chlamydia are not detected by available methods for these diseases, they need to be treated in any case. It is recommended to prescribe an antigonococcal drug (ceftriaxone or ciprofloxacin once) in combination with an antichlamydial drug (azithromycin once or a 7-day course of other drugs). If treatment is ineffective, then re-examination by cultural methods for gonorrhea and chlamydia is necessary. If gonococci are detected, re-treatment after determining sensitivity or if it is impossible to determine it - with a drug from another group. In chlamydia, clinically significant resistance to specific drugs (tetracyclines, erythromycin, azithromycin) has not yet been identified.

Antichlamydial drugs are also effective against genital mycoplasmas in the same doses. Tetracyclines act on both myco- and ureaplasmas. However, recently it has been found that about 10% of ureaplasmas are resistant to tetracyclines, therefore, if treatment of urethritis using doxycycline is ineffective, it is necessary to prescribe erythromycin or azithromycin or ofloxacin.

The species Ureaplasma urealyticum consists of 14 or more serovars, which are divided into 2 biovars. Previously they were called biovar 1 or parvo and biovar 1 or T960. Currently, these biovars are regarded as 2 different species: U.parvum and U.urealyticum, respectively. They vary in prevalence. U.parvum occurs in 81-90%, U.urealyticum in 7-30% of women, and sometimes they are combined - 3-6% of cases. The species U.urealyticum, i.e. former biovar 2 (T960) predominates in women with pelvic inflammatory diseases, pregnancy complications, and is also more often resistant to tetracyclines. The determination of these biovars is carried out for research purposes and is not necessary or economically feasible in routine clinical practice.

Pregnant should be screened for gonorrhea, genital chlamydia, trichomoniasis, bacterial vaginosis and, if detected, receive antibacterial therapy. There is no basis for targeted examination of them for genital mycoplasmas and eradication of these microorganisms. Antibiotics should not be routinely prescribed to prolong pregnancy if there is a threat of termination, except when gonorrhea, trichomoniasis or bacterial vaginosis is detected.

S.V. Sekhin, Research Institute of Antimicrobial Chemotherapy

Ureaplasma and mycoplasma. Questions and answers/h2>

What are ureaplasma and mycoplasma?

• Mycoplasma pneumoniae, which lives in the oropharynx and upper respiratory tract of humans
• and three genital (genital) mycoplasmas that live in the genitourinary system: Human mycoplasma (Mycoplasma hominis)
• Ureaplasma species, which is divided into 2 subspecies (Ureaplasma urealyticum and Ureaplasma parvum)
• Genital mycoplasma (Mycoplasma genitalium)

Recently, pathogenicity (harmful to the body) has been discovered in two more mycoplasmas found in humans. This

• Fermentative mycoplasma (Mycoplasma fermentans), found in the oropharynx
• Penetrating mycoplasma (Mycoplasma penetrans), living in the human genitourinary system.

How common are mycoplasmas in humans?

Ureaplasma sp. is detected in 40-80% of sexually active women who do not complain. In men, the frequency of detection of ureaplasma is lower and amounts to 15-20%. About 20% of newborns are infected with ureaplasma.
Human mycoplasma (Mycoplasma hominis) is detected in 21-53% of sexually active women and 2-5% of men.
About 5% of children over 3 months and 10% of adults who are not sexually active are infected with genital mycoplasmas

How can you become infected with mycoplasma?

Genital mycoplasmas (M. hominis, M. genitalium, Ureaplasma sp., M. penetrans) can be infected only in three ways:

• during sexual contact (including oral-genital contact)
• during transmission of infection from mother to fetus through an infected placenta or during childbirth
• during organ transplantation

Respiratory mycoplasmas (M.pneumoniae, M.fermentans) are transmitted by airborne droplets. Genital mycoplasmas cannot be contracted by visiting swimming pools, toilets or through bed linen.

What diseases can mycoplasmas cause?

Mycoplasmas are often found in healthy people. The reasons why mycoplasmas cause diseases in some people infected with them are still completely unknown. Naturally, most often mycoplasmas cause diseases in people with immunodeficiency caused by HIV infection and with hypogammaglobulinemia (a decrease in the number of certain antibodies), but often mycoplasmas cause diseases in people who do not have immunodeficiency and with normal levels of antibodies.

In women, mycoplasmas can cause the following diseases:

• Cervicitis (inflammation of the cervix) in women is caused by genital mycoplasma (Mycoplasma genitalium)
• Vaginitis (inflammation of the vagina) - there is no proven evidence that genital mycoplasmas cause vaginitis, but ureaplasma and M.hominis are often found in women with bacterial vaginosis
• Pelvic inflammatory diseases (PID) in women - M. hominis was detected in 10% of women with salpingitis; there is also evidence of the possible role of Ureaplasma sp. in the development of PID. and M. genitalium
• Postpartum and post-abortion fever - in approximately 10% of sick women, M.hominis and (or) Ureaplasma sp.
• Pyelonephritis - in 5% of women with pyelonephritis, the cause of the disease is considered to be M.hominis
• Acute urethral syndrome (frequent and uncontrollable urination) in women is often associated with Ureaplasma sp.

In pregnant women, mycoplasmas can lead to the following consequences: possible infection of the placenta, which leads to premature termination of pregnancy, premature birth and the birth of low birth weight newborns.

In both sexes, mycoplasmosis can lead to sexually associated reactive arthritis (joint damage), which is caused by M. fermentans, M. hominis and Ureaplasma sp.

There is evidence of a possible causative role for M. hominis and Ureaplasma sp. in the development of subcutaneous abscesses and ostiomyelitis.
Some studies show a connection between ureaplasma infection and the development of urolithiasis.

Mycoplasmas in newborns

Of particular danger are diseases caused by mycoplasmas in newborns. Infection of a newborn occurs either due to intrauterine infection during pregnancy or childbirth.

The following are associated with genital mycoplasmas in newborns:

• Acute pneumonia (pneumonia) of newborns
• Chronic lung disease
• Bronchopulmonary dysplasia (underdevelopment)
• Bacteremia and sepsis (blood poisoning)
• (inflammation of the meninges)

How are diseases associated with genital mycoplasmas diagnosed?

In the presence of a disease that can be caused by genital mycoplasmas, a cultural study (bacteriological culture for mycoplasma) and a PCR study are carried out.
Determining the presence and amount of antibodies in the blood is not used for diagnosis.

How are diseases associated with genital mycoplasmas treated?

Various antibiotics are used to treat diseases associated with mycoplasmas. The most commonly used are tetracyclines (doxycycline), macrolides (erythromycin, clarithromycin), azalides (azithromycin), fluoroquinolones (ofloxacin, levofloxacin, moxifloxacin). It should be taken into account that different types of mycoplasmas have different sensitivities to different groups of antibiotics.
The effectiveness of the use of drugs that affect the immune system, enzymes, vitamins, local and physiotherapeutic treatment in the treatment of diseases caused by mycoplasmas has not been proven and is not used in developed countries of the world.

How can you protect yourself from infection with genital mycoplasmas?

If you are not infected with mycoplasmas, then you need to take certain measures to prevent infection. The most effective method of protection is the use of a condom.

I was diagnosed with ureaplasma (mycoplasma) using PCR, but I have no signs of the disease. Do I need treatment for ureaplasma (mycoplasma) before conception?

If your sexual partner does not have signs of a disease caused by mycoplasmas and (or) you are not going to change it and (or) are not planning a pregnancy in the near future, then no treatment is prescribed.

I am pregnant and have been diagnosed with ureaplasma (mycoplasma). Do I need treatment for ureaplasma during pregnancy?

Numerous studies have shown that during pregnancy, intrauterine infection and damage to the placenta can occur, which can lead to premature birth and the birth of low-weight newborns, as well as their infection and the development of bronchopulmonary diseases and other complications. Therefore, many doctors prescribe treatment in these cases.

I was diagnosed with a disease associated with ureaplasma (mycoplasma), but my sexual partner has no signs of the disease and the pathogen identified in me is not detected. Does my partner need to be treated for ureaplasma?

No no need. In such cases, some doctors recommend re-examination of sexual partners after a certain period of time (from 2 weeks to a month). During this period, sexual intercourse is prohibited.

I underwent a course of treatment for a disease associated with ureaplasma (mycoplasma) and the pathogen was not detected during control examinations. However, after some time, I again developed symptoms of the disease and the pathogen was discovered. How can this be if during this period I did not have any sexual intercourse?

Most often, re-detection of ureaplasma is due to the fact that complete eradication (disappearance) of the pathogen did not occur and its quantity after treatment decreased to a minimum, which cannot be determined by modern diagnostic methods. After a certain amount of time, the pathogen multiplied, which was manifested by a relapse of the disease.

I took a quantitative test for ureaplasma (mycoplasma) and they were found in a quantity (titer) of less than 10x3. My doctor says that I don’t need treatment, since treatment is prescribed for a higher titer - more than 10x3? Is it true?

The need for treatment is determined not by the quantity (titer) of the detected microorganism, but by the presence or absence of the disease caused by it. If you have signs of illness, you should get treatment. Treatment is also recommended, regardless of the titers identified during quantitative analysis and whether you have signs of the disease, in the following cases: if your sexual partner has signs of a disease caused by ureaplasma (mycoplasmas) and (or) you are going to change your sexual partner and (or) you are planning pregnancy in the near future.

The article used materials from reviews

Ken B Waites, MD, Director of Clinical Microbiology, Professor, Department of Pathology, Division of Laboratory Medicine, University of Alabama at Birmingham

Mycoplasma testing is usually prescribed in combination with other tests that are aimed at detecting other, latent sexually transmitted diseases in the patient. Their list includes the following STDs: trichomoniasis, chlamydia and ureaplasmosis. You can get the above infections in several ways, but the main route of transmission is unprotected sexual contact. An analysis for mycoplasmosis may not give a clearly positive or negative result, since the peculiarity of this infection is its asymptomatic course.

The first signs of infection are urethritis, disturbances in the urination process, burning, discharge, and itching. These are the reasons that should be the impetus for taking a blood test for mycoplasmosis.

Tests for mycoplasmosis and ureaplasma: where to take and get results?

How is a mycoplasma test taken and where to get it? The study is carried out as follows: the patient must, on an empty stomach, visit the laboratory where blood is taken from a vein. You can get tested for mycoplasma and ureaplasma only early in the morning, preferably at least 12 hours after your last meal. Otherwise, this may significantly affect the test results, and the procedure will have to be repeated. No special preparation is required to conduct the study. To test blood for mycoplasma, the polymerase chain reaction or PCR method is usually used, which gives a more detailed interpretation at the end, moreover, its price is very reasonable, and in social clinics a similar test for ureaplasma and mycoplasma can be carried out free of charge. PCR must be taken simultaneously with an enzyme-linked immunosorbent assay, or ELISA. In addition to blood, biological scrapings from the epithelium of the urethra, cervical canal and vagina are taken for research. Decoding the ELISA will help compare the results with the results of PCR, which in turn contributes to a more accurate determination of mycoplasmosis and ureaplasmosis in the blood and epithelial cells of internal organs. It is these two analyzes that have become most widespread in medical practice - due to the high efficiency and low cost of the study.

Mycoplasmosis: blood test to detect pathogen resistance

To determine the resistance of mycoplasma and ureaplasma to a certain type of antibiotics, it is recommended to repeatedly donate blood for testing and culture the bacteria in a specific medium. Based on the results of deciphering this analysis, the doctor can select and prescribe the correct therapy. In contrast to blood diagnostics, the principle of bacterial culture is based on the donation of other biomaterial, and it is based on the artificial cultivation of mycoplasma and ureaplasma pathogens in a special cultural environment. The reliability of deciphering the results of an analysis for mycoplasma largely depends on the equipment and qualifications of the attending physician. In the presence of pathogens ureaplasma and mycoplasma, test results with such a study will be received only after 7-9 days.

Deciphering the analysis for mycoplasma

During direct treatment, the patient will need to undergo repeated blood tests and smears several times, which are taken until the decoding of the results shows a minimum of colony-forming units. After 3-4 weeks, it is necessary to take all tests again; this is done to check the effectiveness of the prescribed therapy and to identify intermediate treatment results. If the final transcript shows the presence of mycoplasmosis and ureaplasmosis, then it is better to take the control test again. A false result in the transcript can be obtained due to incorrect and untimely blood sampling. In order to avoid such mistakes, blood is taken only in specially equipped laboratories.

Specific tests

An analysis for Mycoplasma pneumonia, IgM type antibodies is a test in which blood is taken for diagnosis; this donation of biomaterial serves to identify specific antibodies for Mycoplasma pneumonia. Based on the results, specific immunoglobulins are determined that the defense system produces in the presence of infection. To carry out the test, you need to donate blood from a vein. Afterwards, the resulting serum is sent to the laboratory, where it is examined by ELISA. If the indicators are normal, then the transcript should show negative results. If there are IgM antibodies in the serum, this is the first sign of acute mycoplasmosis. This study is carried out weekly for a month. The price of this test depends on the level of the clinic and diagnostic laboratory. It is important to understand that even after complete recovery, a residual amount of IgM type antibodies remains in the human body. For a complete diagnostic picture, it is also necessary to conduct tests for antibodies of the IgG and IgA types. They will help identify pathogens such as hominis and genitalium. To identify these antibodies, certain parts of pathogen DNA are isolated from serum or mucosal epithelium, and then studied by cloning until a complete picture of the disease is obtained. To obtain reliable data, the biological material taken for diagnostics must be fresh, it cannot be stored for a long time, frozen or subjected to heat treatment. You can donate blood for the test in any laboratory; the speed of obtaining results and the price of the test will vary depending on the complexity of the tests.

Mycoplasmosis is a disease that, when developed, can cause serious complications in the body. Therefore, the appearance of the first symptoms of the disease should serve as a reason to conduct research for the presence of the pathogen in the body. To identify and promptly begin treatment of mycoplasma, tests should be performed as early as possible. The causative agent of the disease is quite dangerous, especially during pregnancy, so contacting a doctor if its signs are detected is mandatory.

Features of the disease

The disease is caused by a microorganism belonging to the class Mycoplasma, which is located on the mucous membranes of the genital organs, and in some cases in the respiratory tract. The peculiarity of this pathogen is that it can remain in the human body for quite a long time asymptomatically, that is, without any manifestations.

Progression of the disease is usually observed after serious gynecological diseases in women, as well as with a significant decrease in the protective functions of the body. It is worth noting that mycoplasmosis often occurs against the background of diseases of the reproductive system such as trichomoniasis, gonorrhea and herpes.

Infection in the overwhelming majority of cases occurs through sexual contact, but the pathogen can also penetrate through household means - through personal hygiene products. Therefore, for preventive purposes, you should strictly observe hygiene and use only your own personal hygiene items. Infection can also occur in utero - and this microorganism is extremely dangerous for the fetus.

The onset of the disease is usually accompanied by mild symptoms, which is why patients do not immediately pay attention to them. The development of mycoplasmosis and the worsening of its symptoms occurs several weeks after infection. The main symptoms of the disease include:

• Women have clear vaginal discharge (scanty or very copious);
• Discharge (clear) from the urethral canal in men;
• Aching pain in the lower abdomen;
• Unpleasant sensations when urinating (burning, itching);
• Painful sensations during sexual intercourse.

In men, mycoplasmosis can also affect the prostate gland, in which case signs of prostatitis begin to appear.

Mycoplasma often causes respiratory and urogenital diseases, since this pathogen is one of the most aggressive among all intracellular organisms. That is why, at the slightest suspicion of this disease, tests are urgently performed that can confirm its presence.

Types of pathogens of mycoplasmosis

The pathogens that cause the disease are microscopic organisms that can initiate infectious and inflammatory processes in the body of men, women and even children. Mycoplasmosis analysis can identify different types of mycoplasmas:

• Pneumonia (Mycoplasma pneumoniae);
• Hominis (Mycoplasma hominis);
• Mycoplasma genitalium;
• Ureaplasma urealyticum.

Of the listed microorganisms, only the first can cause respiratory diseases, while the rest cause genitourinary tract diseases.

Indications for testing

It is mandatory to diagnose mycoplasmosis in the following cases:

• When planning a pregnancy (for both spouses);
• Before carrying out the IVF protocol;
• Before surgical intervention in the pelvic organs;
• If there is a history of miscarriages, miscarriage;
• If the causative agent of the disease is detected in a sexual partner;
• Infertility of unknown origin;
• Frequent manifestations of candidiasis;
• In case of inflammatory processes in the urethra or vagina for unknown reasons;
• The appearance of symptoms of mycoplasmosis.

It is especially important to test for mycoplasma in women during pregnancy, as this infection can lead to miscarriage.

Since the symptoms of mycoplasmosis do not appear immediately, conducting research will help to detect the disease in a timely manner in order to begin treatment.

What tests are performed?

To identify mycoplasmosis, diagnosis is needed, which is carried out using several methods. Three types of research are widely used today:

• Bacteriological;
• Polymerase chain reaction method;
• Enzyme-linked immunosorbent assay (ELISA).

There are other methods, but they are much less effective, so experts have abandoned their use.

Bacteriological method

Otherwise it is called cultural. This method is considered the most accurate test to detect the causative agent of mycoplasmosis in the body. It is carried out by growing microorganisms from the patient’s biological material in special media in laboratory conditions.

“Tank inoculation allows not only to detect mycoplasma, but also to find out the number of microorganisms in a milliliter of the biological material being studied.”

Another advantage of this test is the ability to test how microorganisms respond to different antibiotics to find the optimal treatment for the disease.

Read also on the topic

Mycoplasma in men - causes, signs and treatment

A significant disadvantage of such a study is its duration - culture for mycoplasma can take up to two weeks before obtaining the result. But the reliability of the obtained indicators will be very high. To detect these microorganisms in domestic medicine, they use special tests that can detect mycoplasma hominis and ureaplasma. But not all types of pathogen can be detected as a result of bacteriological examination. Mycoplasma genitalia cannot be determined using bacterial culture, since it grows too slowly (it may take up to 5 months from the moment the smear is taken to obtain reliable results).

Biological material for research in men is obtained from the first portion of urine or by taking a smear from the urethra. Women provide morning urine, a vaginal scraping, or a cervical smear. If the presence of mycoplasma causing respiratory diseases is suspected, sputum is collected for analysis.

The most accurate result will be if there are no foreign impurities in the smear or urine, so biological material is collected from men no earlier than 3 hours after urination, and from women a couple of days before or after the end of menstruation. Also an important condition for the reliability of the results is the absence of treatment with any types of antibiotics in the last month before donating biological material.

Serological studies

Conducting an enzyme immunoassay is also a common way to determine the presence of mycoplasma in the body. This study is based on the detection of special antibodies in the blood - IgA.

Detection of antibodies to mycoplasma in the blood is possible almost immediately after infection. And after complete recovery, they are also present in the ELISA results, but their quantitative indicators in this case do not exceed the norm. It is recommended to carry out the analysis twice to accurately detect the disease, since the production of IgA immunoglobulins requires about 10 days from the moment the pathogen enters the human body. An increase in IgM and IgG titer values ​​indicates the presence of an infectious process that requires treatment.

The presence of IgM in the blood test results indicates an acute course of the infection, and the detection of IgG indicates that the body has previously encountered this microorganism. If both titers are present, they speak of an exacerbation of the chronic process. Thus, when analyzing for mycoplasma hominis, IgG titers indicate that there is no acute course of the disease at the moment.

It is important that the study results are correctly deciphered according to reference values ​​(the norm and deviations from it). Negative results may indicate either the absence of mycoplasma in the blood or a recent infection (less than 10 days), when antibodies have not yet been developed (which is why it is necessary to take the test again). A questionable result indicates a sluggish infection or a disease that is chronic. Positive indicators indicate the presence of a current infection at the moment. If you receive a positive result, experts also recommend undergoing the PCR method or donating a culture test.

Taking blood for analysis does not require special preparation from the patient. Blood is donated in the morning on an empty stomach, and the results of the study will be ready in about 1.5 hours.

But the effectiveness of such research is somewhat reduced due to the peculiarities of the interaction of mycoplasmas with the human body. This pathogen can interact with human cells, which helps them evade the immune response. Because of this, healthy patients may have IgA antibodies, which indicate the presence of the disease, and those patients who have an infection sometimes do not respond to the presence of the microorganism in the blood. That is why this method is used less often than a smear for mycoplasma.

ELISA is usually used for infertility and recurrent miscarriage, some types of complications after childbirth, if the blood test for chlamydia, trichomonas, gonococci, etc. is negative. In such cases, research is most revealing.

Polymerase chain reaction method

This type of research is the most effective, as it allows the detection of mycoplasma DNA in the patient. The PCR method gives positive results much more often than other methods, allowing timely treatment to begin. An important feature of this method is that it detects mycoplasma genitalium - this is the only way to detect the presence of such a microorganism.

Description

Preparation

Indications

Interpretation of results

Description

Determination method PCR with real-time detection.

Material under study Urogenital epithelial cell scraping

Qualitative determination of mycoplasma DNA (Mycoplasma genitalium) in scrapings of epithelial cells of the urogenital tract using polymerase chain reaction (PCR) with real-time detection. Mycoplasma genitalium is a pathogenic microorganism that causes diseases of the genitourinary tract. Mycoplasmas are transmitted through sexual contact and can cause nongonococcal urethritis and prostatitis, pelvic inflammatory diseases, pathologies of pregnancy and the fetus, and infertility in women and men.

Analytical indicators:

• the determined fragment is a specific section of Mycoplasma genitalium DNA;
• detection specificity - 100%;
• sensitivity of the analysis - 100 copies of Mycoplasma genitalium DNA per sample.

Preparation

It is advisable to conduct examinations of women in the first half of the menstrual cycle, not earlier than the 5th day. Examination in the second half of the cycle is acceptable, no later than 5 days before the expected start of menstruation. If there are severe symptoms of inflammation, the material is taken on the day of treatment. The day before and on the day of the examination, the patient is not recommended to douche the vagina. It is not recommended to take biomaterial during antibacterial therapy (general / local) and during menses, earlier than 24-48 hours after sexual intercourse, intravaginal ultrasound and colposcopy. It is recommended to take the material no earlier than 14 days after the use of antibacterial drugs and local antiseptics, and no earlier than 1 month after the use of oral antibiotics. If a scraping is taken from the urethra for research, the material is collected before or no earlier than 2 to 3 hours after urination.

Indications for use

• Establishing the etiology of a chronic infectious process of the urogenital tract.
• An erased picture of inflammation of the genitourinary system.
• Pregnancy.
• Ectopic pregnancy.
• Infertility.
• Weakening of the immune system.
• Monitoring the effectiveness of antibiotic therapy (not earlier than a month after taking antibacterial drugs).
• Preventive screening studies (to exclude the possibility of asymptomatic and latent infection).

Interpretation of results

Interpretation of research results contains information for the attending physician and is not a diagnosis. The information in this section should not be used for self-diagnosis or self-treatment. The doctor makes an accurate diagnosis using both the results of this examination and the necessary information from other sources: medical history, results of other examinations, etc.

The test is qualitative. The result is given in terms of “detected” or “not detected”.

• “detected”: a DNA fragment specific for Mycoplasma genitalium was found in the analyzed sample of biological material: infection with Mycoplasma genitalium;
• “not detected”: no DNA fragments specific to Mycoplasma genitalium were found in the analyzed sample of biological material or the concentration of the pathogen in the sample is below the sensitivity limit of the test.

5 004

Mycoplasmosis and ureaplasmosis are diseases that do not have any specific symptoms unique to these infections. Therefore, laboratory research methods are decisive for making a diagnosis.

In order to reliably establish the diagnosis " urogenital mycoplasmosis" or " ureaplasmosis", 2 prerequisites are required:

1. The presence of an inflammatory process of the urogenital system.
2. The presence of the causative agent mycoplasma or ureplasma, proven by laboratory methods. In this case, other possible pathogens should be absent.

Who should be tested for chlamydia first?

• Women and men suffering from infertility of unknown cause for more than 2 years.
• Women with chronic inflammatory diseases of the genitourinary system of unknown etiology (especially when planning pregnancy).
• Pregnant women who have previously had spontaneous miscarriages, premature birth, polyhydramnios, etc.
• Pregnant women with an unfavorable course of this pregnancy.
• Patients with urolithiasis and pyelonephritis, because they are a high-risk group for mycoplasmosis.
• Patients with prostatitis, long-term urethritis.
• Before gynecological and urological operations.

Is it necessary to undergo examination if there are no symptoms of the disease?
Only necessary in the above cases. At the same time, there is no need to conduct special examinations for myco- and ureaplasmosis during preparation or during pregnancy if nothing bothers the woman. The fact is that normally these bacteria are found in approximately 50% of women, so even in the absence of complaints they can be detected, but it is not worth treating asymptomatic carriage of these microbes.
In all other cases, there is also no need to take tests for myco- and ureaplasmosis “just in case”.

What are they researching?
To detect myco- and ureaplasmas, it is necessary to collect material. This can be a scraping containing cells of a diseased organ - vagina, cervix, prostate secretion, scraping from the urethra, conjunctiva of the eye. Such material can also be blood, urine and semen in men.

What tests are prescribed for mycoplasmosis and ureaplasmosis?
For myco- and ureaplasmosis, the following tests are most appropriate:
1. Polymerase chain reaction (PCR) - determination of pathogen DNA.
2. Enzyme-linked immunosorbent assay (ELISA) – determination of antibodies to the pathogen.
3. Microbiological research (cultural method) - finding the pathogen itself.

1. Polymerase chain reaction (PCR).

• The method is based on the detection of the genetic material of the pathogen in the sample being studied. Using PCR, a specific section or DNA fragment of myco- and ureaplasmas is detected in the material under study, therefore, in comparison with other methods, it is impossible to confuse them with some other infection.
• PCR allows you to detect the pathogen even in latent, chronic and asymptomatic forms of infection, when other research methods are not informative.
• Using PCR, it is possible to detect mycoplasma and ureaplasma even in the incubation period, when there are no clinical manifestations of mycoplasmosis.
• PCR analysis requires very little material, and the results are ready in 1-2 days.
• When diagnosing a primary infection, it is more informative to identify this infection in the places of initial localization, i.e. The material should be scrapings from the genital tract.
• False-positive results are possible in PCR analysis. This can happen if the study was conducted earlier than a month after a course of antibiotic therapy. The fact is that when identifying a fragment of mycoplasma DNA, it is impossible to assess whether it is a dead or viable microbial cell. In this case, the viability of mycoplasmas is assessed using a microbiological method. If the bacterium is not viable, then, despite the presence of a DNA fragment, the microbial cells will not grow in the cell culture.
• False-negative results are also possible if the collection process, transportation of the material, and the analysis itself are disrupted.
• To date, the accuracy of this method, when performed correctly, is the highest - up to 100%.

If the PCR test for mycoplasma is positive, but there are no symptoms of mycoplasmosis, it is necessary to conduct other research methods.

2. Enzyme-linked immunosorbent assay (ELISA)– determination of antibodies to the pathogen.

• ELISA is a method for indirect detection of bacteria, i.e. the pathogen is not detected directly, but specific antibodies (IgG, IgA, IgM) to it and the body’s reaction to its introduction are determined.
• ELISA allows you to determine at what stage the disease is - acute or chronic, and evaluate the effectiveness of the treatment.
• Specific Ig A are produced during fresh infection, IgM indicate the presence of an active infection. The presence of only IgG without IgM indicates a past infection, which is currently absent or a carrier state. For more information about evaluating the results of the ELISA analysis, see the article “”.
• The accuracy of ELISA is about 80%. This is due to the fact that antibodies to chlamydia can be present in healthy people as a result of a previous illness, and can also be detected in respiratory and other types of mycoplasma infections.

3. Microbiological examination (culture method) with determination of sensitivity to antibiotics.

• The essence of this method is that the material under study is sown on a special medium and grown. Then the pathogen is identified based on its growth pattern and other characteristics. The cultural method allows not only to identify viable myco- and ureaplasmas, but also to select an antibiotic to which they are sensitive.
• Diagnosing mycoplasmosis is quite difficult, because... mycoplasmas can be a component of the natural microflora of the genitourinary organs in healthy individuals. The presence of myco- and ureaplasmas in test results is not a disease. For an accurate diagnosis, it is necessary to know the number of bacteria in the genitourinary organs.
• Only the cultural method makes it possible to determine the amount of the pathogen in the test material, and therefore to distinguish asymptomatic carriage of myco- and ureaplasmas from the corresponding diseases. To do this, count the number of colonies grown on the medium, which are called colony-forming units (CFU). This number indicates the number of living bacteria that can reproduce to form colonies.
• With asymptomatic healthy carriage of myco- or ureaplasmas, less than 104 CFU/ml is determined. If the disease is present, the number of mycoplasma or ureaplasma colonies in the test material will be more than 104 CFU/ml.
• The accuracy of identifying bacteria with this method reaches 95%.
• For more information about the method, see the article “?”.

So, all these 3 methods are quite accurate, but they are all complementary.
Why? To do this, you need to clearly understand the capabilities of each method.

Possibilities and limitations of laboratory tests.

• ELISA: allows you to assess the state of immunity and the body’s response to the pathogen, indirectly indicates the presence of myco- or ureaplasmas in the entire body, but does not indicate the specific affected organ. Allows you to evaluate the effectiveness of antibiotic therapy. But with a weak immune response, for example, in patients with immunodeficiencies, ELISA is not informative.
• PCR: allows you to accurately determine the location of the pathogen, but this is not always available (for example, in the ovaries). Allows you to detect the pathogen even in latent, chronic and asymptomatic forms, as well as during the incubation period. Characterized by the highest accuracy of pathogen identification. It does not allow assessing the effectiveness of antibiotic therapy, distinguishing carriage from disease, or assessing the viability of the pathogen.
• Culture method: allows you to identify viable bacteria, determine their number, evaluate the effectiveness of antibiotic therapy, and distinguish carriage from disease. Does not evaluate the body's response to the pathogen.

conclusions

• There is not a single method that would detect mycoplasmas in 100% of cases. Therefore, laboratory diagnostics must include at least two methods.
• If it is impossible to take material from the organ being studied, ELISA is used.
• To assess the effectiveness of treatment, a culture method is used. If it is not possible, use ELISA.
• To determine the stage of the disease - ELISA.
• In patients with immunodeficiencies, ELISA is not informative; PCR and culture methods are used.
• You should not rely too much on the results of determining the sensitivity of mycoplasmas to antibiotics. After all, as is known, microorganisms behave differently in a test tube (in vitro) and in a living organism (in vivo).