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Journal : Makara Journal of Science

Construction of pcDNA3.1 Vector Encoding RpfD Gene of Mycobacterium tuberculosis Rakhmawati, Aprilia; Rukmana, Andriansjah; Karuniawati, Anis
Makara Journal of Science Vol. 22, No. 3
Publisher : UI Scholars Hub

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Abstract

Tuberculosis (TB) is an infectious diseasecaused by Mycobacterium tuberculosis (M. tuberculosis). TB is still a major health problem. The Bacillus Calmette-Guérin (BCG) vaccineis the only one available for TB and is known to confer variable levels of protection. Because of thisvariability, a new vaccine is needed to control TB. Proteins secreted by M.tuberculosisare known to induce protective immunity. Within the genome of M. tuberculosis, there is a family of proteins called resuscitation promoting factor (Rpf), which playsa role in the reactivation of M. tuberculosis. RpfD is amember of the Rpf family that has been shown to be immunogenic, makingitsuitable for use as a TB vaccine. The rpfD gene of the M. tuberculosis Beijing strain from the bacterial stock of the Department of Microbiologyat the Medical Facultyof theUniversitas Indonesia was amplified using polymerase chain reaction (PCR) and then insertedintothemammalian expression vector pcDNA3.1(+). Then, the pcDNA3.1(+)-rpfD vector was transformed to Escherichia coli DH5α. A 465-bp target fragment was obtained, and the accuracy ofthecloning was confirmed using colony PCR, restriction enzyme digestion, and sequencing. We expect that this recombinant plasmid will induce immunity in future animal models and thus will prove itself to be a candidate for an M. tuberculosis vaccine.
Efficacy of Tuberculosis Vaccine Candidate pcDNA3.1-rpfB in Inhibiting the Growth of Mycobacterium tuberculosis In Vitro with Mycobacterial Growth Inhibition Assay Pujilestari, Ratih; Rukmana, Andriansjah; Karuniawati, Anis
Makara Journal of Science Vol. 26, No. 1
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Abstract

Tuberculosis (TB) is caused by Mycobacterium tuberculosis (Mtb). Bacille Calmette-Guérin (BCG) is the only licensed vaccine against TB, and it is 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 induces immune responses in mice, but its potency is unknown. The gold standard for potency testing of TB vaccine is the challenge method. The BSL3 animal laboratory for the challenge method is currently unavailable at FKUI. Therefore, mycobacterial growth inhibition assay (MGIA) was used as a preliminary test before the in vivo challenge test was conducted. The principle of MGIA is to reculture Mtb in a Mycobacteria Growth Indicator Tube (MGITTM) from co-cultured Mtb with mammalian cells that have been previously treated with pcDNA3.1-rpfB, pcDNA3.1 (negative control), and BCG (positive control). MGITTM shows the time to positivity, which is the time that has lapsed until a positive growth of Mtb is detected. In addition, measurements of interferon (IFN)γ levels by enzyme-linked immunosorbent assay were carried out. This study concluded that pcDNA3.1-rpfB can inhibit the growth of Mtb in vitro and showed no statistical difference from BCG. The IFNγ levels from co-culturing did not correlate with the level of inhibition of the growth of Mtb in vitro.
Co-Authors . Andriansjah Ade Dharmawan Amalia Almira Amarila Malik Amin Soebandrio Andi Annisa Rusyda Khafiyani Andi Yasmon Andra Aswar Andra Aswar, Andra Angky Budianti Anwar S. Ibrahim Aprilia Rakhmawati Ardiana Kusumaningrum Ari Fahrial Syam Ari Prayitno Aria Kekalih Aryo Tedjo Asmarani Kusumawati Asmarani Kusumawati Augustine Natasha Aulia Rizki Bachti Alisyahbana Badriul Hegar Badriul Hegar Bambang Madiyono Burhanuddin Iskandar Ceva W. Pitoyo Chairunissa T. Rizal Cleopas Martin Rumende Cleopas Martin Rumende Delly Chipta Lestari Delly Chipta Lestari Delly Chipta Lestari Delly Chipta Lestari Delly Chipta Lestari Dimas Seto Prasetyo Dwiana Ocviyanti E. Risdiyani Em Yunir Em Yunir, Em Erwin Christian Susanto Esthika Dewiasty Esthika Dewiasty, Esthika Falasiva, Rezyta Fariz Nurwidya Febrianti, Tati Febriyana, Dwi Fitriah, Laela Futihati Ruhama Zulfa Gestina Aliska Gortap Sihotang Hana Paraswati Putri Handaya, Adeodatus Y. Hasanah, Lili Hindra Irawan Satari Hindra Irawan Satari Ida Parwati Ika Ningsih Indah S. Widyahening Iswanto, J. Joyce Bratanata Joyce Bratanata, Joyce Kambang Sariadji Khie Chen Kuntjoro Harimurti Lalisang, Toar JM. Linosefa Linosefa Manuel Lamberto Willem Mboeik Martin Hartiningsih Mboeik, Manuel Lamberto Willem Moehario, Lucky Hartati Muhammad K Azwar, Muhammad K Mulya Rahma Karyanti, Mulya Rahma Mulyadi M. Djer Muziasari, Windi Nabila Maudy Salma Nadia Maretti Nasution, Safruddin Nelly Puspandari Nie Nie Nina Dwi Putri Nuraini I Susanti Permatasari, Tri Wijayanti Pratama Wicaksana Pratiwi Pujilestari Sudarmono Pratiwi Sudharmono Pratiwi Sudharmono, Pratiwi Prawoto Prawoto Prawoto Prawoto Pujilestari, Ratih Purwantyastuti Purwantyastuti Puspandasari, Nelly Rakhmawati, Aprilia Resti H. Lestari Reynaldo Reynaldo Reynaldo Reynaldo, Reynaldo Rezyta Falasiva Riamin Sitorus Rianto Setiabudi, Rianto Rida Tiffarent Rosdiana Irawati Rudyanto Sedono Rudyanto Sedono Rudyanto Sedono, Rudyanto S. Nilawati Santya Fatma Dewi Sari Wiraswasty Saunar, Rofy S. Shindy Claudya Aprianti Simanjuntak, Glory Gelarich Sitorus, Truly Panca SJATHA, FITHRIYAH Subari A. Riyanto Sudigdo Sastroasmoro Sukman T. Putra Sunarno Sunarno Susan M. Noor Susan M. Noor, Susan Susan Maphilindawati Noor Susanti, Nuraini I Susanti, Nuraini I Susanto, Erwin Christian Sweety Pribadi SYADZA RHIZKY PUTRI AKHMAD T. M. Sudiro Teguh Harjono Karjadi Teguh Harjono Karjadi, Teguh Harjono TETSUHIRO MATSUZAWA Thomas Robertus Tjampakasari, Conny Riana TOHRU GONOI Tony Loho Trisni U Dewi, Trisni U Truely Panca Sitorus Usman, Nurhayat Weaver, Tom Wia Melia Windi Muziasari Yefta Moenadjat Yeva Rosana Yulia R. Saharman Yulia Rosa Saharman