Tesis

Potensi Kandidat Vaksin Tuberkulosis pcDNA3.1rpfB dalam Menghambat Pertumbuhan Mycobacterium tuberculosis secara In-Vitro = Potency of Tuberculosis Vaccine Candidate pcDNA3.1-rpfB in Inhibiting the Growth of Mycobacterium tuberculosis In-Vitro.

Tuberkulosis (TB) adalah penyakit akibat infeksi Mycobacterium tuberculosis (M. tuberculosis). Bacille Calmette-Guérin (BCG) adalah satusatunya vaksin berlisensi untuk melawan TB, terbukti efektif pada anak-anak tetapi tidak pada orang dewasa. Tim Peneliti Vaksin Departemen Mikrobiologi FKUI telah mengembangkan kandidat vaksin TB berbasis vaksin DNA pcDNA3.1-rpfB. Kandidat vaksin ini terbukti dapat menginduksi respon imun humoral dan seluler pada mencit tetapi belum diketahui potensinya. Baku emas uji potensi vaksin TB adalah metode tantang. Fasilitas laboratorium hewan BSL3 untuk metode tantang tidak tersedia di FKUI. Oleh karena itu, dalam penelitian ini digunakan Mycobacterial Growth Inhibition Assay (MGIA). Prinsip MGIA adalah merekultur M. tuberculosis pada Mycobacteria Growth Indicator Tube (MGIT TM ) dari hasil co-culture M. tuberculosis dengan sel mamalia yang sebelumnya telah diberi perlakuan dengan pcDNA3.1-rpfB, pcDNA3.1 (kontrol negatif), dan BCG (kontrol positif). MGIT TM akan memberikan nilai Time to Positivity, yaitu waktu yang dibutuhkan hingga terdeteksi positif pertumbuhan M. tuberculosis. Selain itu, dilakukan juga pengukuran kadar IFNγ terhadap supernatan hasil co-culture. Berdasarkan hasil penelitian ini disimpulkan bahwa pcDNA3.1-rpfB dapat menghambat pertumbuhan M. tuberculosis secara in-vitro yang tidak berbeda dengan BCG. Kadar IFNγ hasil co-culture M. tuberculosis dengan sel mamalia tidak berkorelasi dengan tingkat penghambatan pertumbuhan M. tuberculosis secara in-vitro.
Kata kunci: Mycobacterial Growth Inhibition Assay, Mycobacterium tuberculosis, resuscitation-promoting factor B (rpfB), vaksin.


Tuberculosis (TB) is caused by Mycobacterium tuberculosis (M. tuberculosis). Bacille Calmette-Guérin (BCG) is the only licensed vaccine against TB, effective in children but not in adults. The Vaccine Research Team, Department of Microbiology FKUI has developed a DNA based TB vaccine candidate pcDNA3.1-rpfB. This candidate has been shown to induce immune responses in mice but its potency is unknown. The gold standard for potency testing of TB vaccine is a challenge method. BSL3 animal laboratory for the challenge method are not available at FKUI. Therefore, this study used Mycobacterial Growth Inhibition Assay (MGIA). The principle of MGIA is to reculture M. tuberculosis in Mycobacteria Growth Indicator Tube (MGIT TM ) from co-cultured M. tuberculosis with mammalian cells that have been previously treated with pcDNA3.1-rpfB, pcDNA3.1 (negative control), and BCG (positive control). MGIT TM will show Time to Positivity, time taken until a positive growth of M. tuberculosis is detected. In addition, measurements of IFNγ levels by ELISA were also carried out. This study concluded that pcDNA3.1-rpfB could inhibit the growth of M. tuberculosis in-vitro which was not different from BCG. IFNγ levels from co-culturing did not correlate with the level of inhibition of the growth of M. tuberculosis in-vitro.
Keywords: Mycobacterial Growth Inhibition Assay (MGIA), Mycobacterium tuberculosis, resuscitation-promoting factor B (rpfB), vaccine.