It is used for the formulation of the intradermal Mantoux test with various doses of tuberculin, for dermal and subcutaneous tuberculin samples (graduated skin test,subcutaneous test, determination of tuberculin subcutaneous and intradermal titer, eosinophilic tuberculin test, hemo-protein-tuberculin test, and others).
Tuberculin samples are placed in the sitting position for patients. In emotionally labile people, injection can cause fainting.
Dilutions of purified tuberculin are prepared as follows: the ampoule is wiped with gauze moistened with 70 ° ethyl alcohol, then the ampulla neck is filed with an ampoule knife and broken off. Similarly, open the ampoule with the solvent.
For intradermal samples, purified tuberculin is diluted in aseptic conditions as follows: the contents of the ampoule are diluted with 1 ml of the solvent and thus the main dilution is 50,000 TE in 1 ml. The drug should dissolve within 1 min, be transparent and colorless.
The 1 st dilution, corresponding to 1000 TE in 0.1 ml, is prepared by adding 4 ml of solvent - carbolized sodium chloride solution to the ampoule with the main dilution.
All the subsequent dilutions are prepared by diluting the previous one in a 1:10 ratio, mixing them thoroughly. For example, the 2nd dilution: add 1 ml of the solvent to 1 ml of the 1st dilution, which corresponds to 100 TE in 0.1 ml.Similarly, from the 2nd dilution, prepare the 3rd (10 TE in 0.1 ml), etc.
To obtain 2 TE in 0.1 ml to 1 ml of the 3rd dilution of tuberculin, 4 ml of the solvent should be added.
Dilutions of tuberculin are preserved under aseptic conditions for no more than 2 hours.
Tuberculin samples using purified tuberculin are placed on the order of the phthisiatrician. The presentation and evaluation of tuberculin samples is performed by a doctor or specially trained nurse under the supervision of a doctor.
1. Intradermal Mantoux Test
Intradermal Mantoux test is performed strictly aseptically as follows: open the ampoule and prepare the appropriate dilution as described above. To place intradermal tests, use disposable tuberculin syringes with thin needles. Do not use syringes and needles with expired use for intradermal tests, insulin syringes. In a syringe with a needle No. 0845, 0.2 ml is collected, i.e. 2 doses of dilution, put a thin needle, release the solution to a label of 0.1 ml in sterile cotton tampon. The skin of the middle third of the inner surface of the forearm is treated with 70 ° ethyl alcohol and dried with sterile cotton.The medical worker with his left hand fixes the skin of the patient's forearm from below so that it is stretched on the inner surface. A thin needle is cut upward into the upper layers of the skin parallel to its surface - intradermally. After inserting the needle into the skin, the syringe is injected strictly by dividing the scale with 0.1 ml of the preparation. With the correct technique of introduction, a papule of a whitish color is formed in the skin in the form of a lemon crust, the size 7-10 mm in diameter. For each examinee, use a separate sterile syringe and needle.
2. Definition of tuberculin intradermal caption
Purified tuberculin in dilutions of 1 TE in 0.1 ml, 0.1 TE in 0.1 ml and 0.01 TE in 0.1 ml (prepared as described above) is administered by the Mantoux method simultaneously in one forearm. If the results are negative, the sample with more concentrated tuberculin (3rd dilution) is repeated on the other hand not earlier than 96 hours. If there is no reaction to the 3rd dilution of tuberculin, the samples with the 2nd and 1st dilutions are successively delivered after 36 hours each , in different forearms.
Titration is completed upon reaching a positive reaction to the intradermal test (papule of at least 5 mm in diameter) obtained for the smallest dilution of tuberculin.
3.Graduated skin test
Graduated skin test is performed simultaneously with solutions of purified tuberculin 100%, 25%, 5% and 1%.
For the preparation of 100% tuberculin containing 100 000 TE in 1.0 ml, 2 ampoules of the preparation are opened, as described above, the contents of the ampoules are diluted sequentially in 1 ml of the solvent.
For the preparation of solutions of 25% (25000 TE) of tuberculin in a sterile 1.5 mL vial poured karbolizirovannogo solvent and added to 100 ml of 0.5% solution of tuberculin.
For the preparation of tuberculin 5% (5000 TE) of the concentration in the bottle are poured 2 ml of solvent and add there 0.5 ml 25% solution of tuberculin.
To obtain 1% (1000 TE) concentration in a bottle with 2 ml of solvent, add 0.5 ml of a 5% solution of tuberculin.
Drops with the prepared tuberculin solution are pipetted onto the prepared forearm skin (see above) at a distance of 2-3 cm from each other. In the upper part of the forearm, a drop of 100% solution is applied, and below it drops of decreasing concentration. Even lower as a control, a drop of solvent is applied. For each solution, separate labeled pipettes are used. The skin is fixed as in the setting of the intradermal test (see above).Then, the invasive pen is broken by the integrity of the surface layers of the skin in the form of a 5 mm long scratch drawn through a drop of tuberculin in the direction of the longitudinal axis of the arm. The flat side of the pen is produced by rubbing tuberculin (2-3 times). Scarification is carried out first through a drop of the solvent, then sequentially through drops of 1%, 5%, 25% and 100% solutions of tuberculin. The scarified area of the skin is left open for 5 minutes to dry the drops of tuberculin. A sterile pen is used for each subject.
To determine tuberculin subcutaneous titer, subcutaneous test, for eosinophilic tuberculin, hemo-protein-tuberculin and other samples, purified tuberculin and its dilutions are used according to special methods.
Profitability Analysis
Results of cutaneous and intradermal tuberculin samples are evaluated after 72 hours, by measuring the size of the infiltrate (papules) in mm. Transparent ruler with millimetric divisions measure and record the transverse (in relation to the axis of the forearm) the size of the infiltrate. Hyperemia is only considered if there is no infiltrate.
When setting Mantoux test, the reaction is considered:
- negative - in the complete absence of infiltration (papules) and hyperemia or in the presence of a knock-off reaction (up to 1 mm);
- questionable - with infiltrate size (2-4 mm) or only hyperemia of any size without infiltration;
- positive - in the presence of an infiltrate with a diameter of 5 mm or more;
- hyperergic - in children and adolescents is considered a reaction with a diameter of the infiltrate of 17 mm and more, in adults - 21 mm and more, as well as vesiculonecrotic reactions regardless of the size of the infiltrate with lymphangitis or without it.
Response to the introduction of tuberculin
The reaction of delayed type hypersensitivity (HRT) depends on the level of specific reactivity of the organism. The peak of HRT comes through 48-72 hours, by this time the manifestation of a local reaction of an immediate type (hyperemia) disappears.