The thesis is devoted to identify theoretical and practical justification, summarized research results of development, implementation and harmonization with the EU requirements «PPD Tuberculin Mammalian purified» drug for the allergic reactions of the macroorganism.
Selected by breeding, the sealed production strains M. bovis Valleе KMIEV-9 and M. bovis Vallеe KMIEV-9KM appeared to be highly proteinogenic, they comply with the requirements of Council Directive 97/12 (dated March 17, 1997) and used during the production of experimental production series PPD-tuberculin for purified mammalian. The mass fraction of protein in a sample of tuberculin made from the strain M. bovis Valleе KMIEV-9 is (0,89 ± 0,1) mg / cm3, while in the sample of tuberculin of the strain M. bovis Valleе, KMIEV-9KM it is significantly higher (p <0 , 05) and it comes up to (1.20 ± 0.2) mg / cm3. The production strain of mycobacterium M. bovis Valleе KMIEV-9KM during cultivation on a synthetic nutrient medium of Soton HB or KF allows to accelerate the growth and increase the accumulation of bacterial mass of mycobacteria from one vial to (16,1 ± 0,2) mg and allows to increase the output of tuberculin accordingly to (1.20 ± 0.1) mg / cm3. For the adaptation, selection and accumulation of production strains bacterial mass, Soton KF and Soton-KhB nutrient media were developed, on which the M. bovis culture begins to grow (4.2 ± 1.1) days earlier and gives more accumulation of bacterial mass (microbial film is formed (4,1 ± 0,9) days earlier, in comparison with an initial strain).
New technological methods of PPD-tuberculin manufacturing have been developed: the method of obtaining tuberculin from culture fluid and bacterial mass of mycobacteria, and the method of manufacturing PPD-tuberculin using membrane microfiltration and ultracentrifugation methods, which allowed to obtain highly active and specific diagnostic allergen.
When making tuberculin with the use of membrane microfiltration technology and ultracentrifugation at 14,000 rpm., some product characteristics significantly increases: the protein yield after trichloroacetic acid precipitation (p <0,05) to (8,6 ± 0,5) g, the yield of tuberculin from 1 liter of medium (0,7 ± 0,1) g and the mass fraction of protein to (0.6 ± 0.1) mg / cm3. The drug is guaranteed to be sterile and highly purified. To conduct control studies of experimental and production series PPD-tuberculin for mammals, a method has been developed for determining the activity of purified tuberculin (MAP) for mammals in animals suspected of avirulent cultures of M. bovis, which involves the use of inactivated or attenuated mycobacteria.
The developed method is specific and safe for the health of animals and humans and prevents the proliferation of living mycobacteria in nature. Experimental research series No. 1 (E) and No. 2 (E) PPD-Tuberculin for purified mammals have showed high diagnostic activity and specificity.
The activity of the tested series of tuberculins No. 1 (E) and No. 2 (E) was 49,000 IU / mg and 47000 IU / mg, respectively, which complies with DSTU 4664:2006 and Council Directive 97/12 of 17 March 1997, according to this Directive, the tuberculinization of animals is carried out using PPD or HCSM tuberculins, which have to be manufactured from M. bovis strains AN5 or Valleе.
