2 research outputs found
KIT DIAGNOSTIK DIFTERI BERBASIS HIBRIDISASI DNA
No full textIntroduction: Diphtheria is an upper respiratory tract infection caused by Corynebacterium diphtheria bacterium. The recent diphtheria outbreak caused a large number of deaths in rapid period. This is due to the absence of a fast and precise method of diphtheria diagnosis. Current diagnosis practices aredone by cultivating bacteria which takes a relatively long time, which is 3 to 5 days, while the patients may already develop severe infection by the time the results are accessible. Therefore, a sensitive, easy and fast diphtheria diagnostic method is needed so that patients suspected of being infected can betreated quicker.Methods: This advertorial is written based on literature study method.Result: The diphtheria diagnostic kit was designed based on the DNA hybridization method by using DNA microarray. There are probes on the microarray plate in the form of a single stranded DNA fragment that is complement to the unique area of gene that encodes diphtheria toxin. The test sample wasobtained from the preparation of the patient's throat smear and labeled with a fluorescence marker before dropped onto the plate. Thus, if there is diphtheria DNA in the patient's sample, hybridization will occur between diphtheria DNA and the complementary DNA probe. The occurrence of hybridization will be characterized by fluorescence as an indicator of whether patients have diphtheria or not. The application of this diagnostic kit can shorten the diagnosis of C. diphtheria to less than 24 hours .Conclusion: Diphtheria diagnostic kit based on DNA hybridization can be made and applied.Pendahuluan: Difteri merupakan penyakit infeksi saluran pernapasan atas yang disebabkan oleh bakteri Corynebacterium diphtheria. Kemunculan kembali wabah difteri menyebabkan kematian dalam jumlah banyak dan dalam kurun waktu cepat. Hal ini diakibatkan karena tidak adanya metode diagnosisdifteri yang cepat dan tepat. Metode diagnosis umum yang saat ini dilakukan adalah kultivasi bakteri dan metode ini membutuhkan waktu yang cukup lama, yaitu 3 hingga 5 hari, dengan kondisi pasien yang mungkin sudah sangat parah ketika hasil diagnosis tersebut dapat dirilis. Oleh karena itu,dibutuhkan metode diagnosis difteri yang sensitif, mudah dan cepat sehingga pasien yang diduga terjangkit dapat ditangani dengan lebih cepat.Metode: Advertorial ini ditulis berdasarkan metode penelusuran pustaka.Hasil: Kit diagnostik difteri dirancang berdasarkan metode hibridisasi DNA menggunakan metode DNA microarray. Pada pelat microarray terdapat probe berupa fragmen DNA untai tunggal yang merupakan komplemen dari daerah unik berupa gen pengkode protein toksin difteri. Sampel uji diperoleh daripreparasi sampel apusan tenggorokan pasien kemudian dilabeli dengan penanda fluoresensi dan diteteskan ke pelat. Dengan demikian, jika terdapat DNA difteri di dalam sampel maka akan terjadi hibridisasi antara DNA difteri dengan probe. Terjadinya hibridisasi akan ditandai dengan fluoresensi yang merupakan indikator pasien terjangkit difteri atau tidak. Pengaplikasian kit diagnostik ini dapat mempersingkat proses diagnosis C. diphtheria menjadi kurang dari 24 jam.Kesimpulan: Kit Diagnostik Difteri berbasis hibridisasi DNA dapat dibuat dan diterapkan
In Silico Study, Design, and Expression of an Intranasal Dual Chimeric Vaccine for Indonesian-Based Norovirus GII-2 and Hepatitis B
No full textHepatitis B virus (HBV) remains an important healthcare challenge, leading to liver diseases like cirrhosis and cancer. In response, we created a prophylactic and therapeutic HBV vaccine by integrating HBcAg and HBsAg from HBV genotype B into Norovirus (NoV) GII.2 P domain (PdomGII.2-HBV) for enhanced intranasal delivery. This vaccine also aimed to simultaneously prevent NoV infection, which causes gastroenteritis. Since the selected HBV epitopes have undergone extensive research and are tailored to the Indonesian population, this study focused on identifying NoV epitopes and assessing T cell epitopes coverage of the PdomGII.2-HBV for the Indonesian population. Following that, we expressed the PdomGII.2-HBV protein using Escherichia coli BL21(DE3) and employed a gentle solubilization technique for protein purification. Our in-silico analysis identified two B cell epitopes, along with 15 CD4+T cell epitopes and 35 CD8+T cell epitopes within the GII.2 P domain. These T cell epitopes cover 100% of the Javanese-Sundanese population\u27s HLA allele variations, which constituted the largest demographic group in Indonesia. Subsequently, we successfully purified the presumed PdomGII.2-HBV protein, revealing a molecular weight of 39.5 kDa. Following the successful expression and purification of the presumed PdomGII.2-HBV protein, it is evident that this vaccine design has significant potential, warranting further study
