OPTIMIZATION OF IN HOUSE POLYMERASE CHAIN REACTION FOR HIGH RISK HUMAN PAPILLOMAVIRUS (HPV 18) E1 GENE-BASED DETECTION
DOI:
https://doi.org/10.61841/q64t4889Keywords:
Human Papillomavirus, Polymerase Chain Reaction, E1 geneAbstract
Human Papillomavirus is a virus that can infect human and a causative agent for cervical cancer. This disease mostly caused by HPV type 16 and 18. Polymerase Chain Reaction (PCR) is one of the molecular biology method can be used to detect HPV in sample. Gen E1 in HPV genome has a role in virus replication and transcription, and relatively conserved. This gene can be used in HPV genotyping and detection. This research aim is to find the PCR optimal reaction for HPV type 18 detection. The E1 gene data from the National Center for Biotechnology Information (NCBI) was used in designing primer with Primer-BLAST NCBI. Selection of the primers was done by in silico analyzes using DINAmelt and Mfold. Optimization of annealing temperature and primer concentration was done for selected primers. The result showed that optimal annealing temperature is 59oC with optimal primer concentration is 600 nM
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References
Bernard, Hans-Ulrich. Robert D. Burk, Zigui Chen, Koenraad van Doorslaer, Harald zur Hausen & Ethel-Michele de
Villiers. (2010). Classification of Papillomaviruses (PVs) Based on 189 PV Types and Proposal of Taxonomic
Amandments. National Institutes of Health Public Access Virology, 401(1), 70–79.
Bioline. (2019). MyTaqTM HS DNA Polymerase. Retrieved from
https://www.bioline.com/downloads/dl/file/id/2725/mytaq_hs_dna_polymerase_v7-manual.pdf
Bruni, L., Albero G., Serrano B., Mena M., Gomez D., Munoz J., Bosch FX, de Sanjose S. ICO/IARC Information Centre
on HPV and Cancer (HPV Information Centre). (2019). Human Papillomavirus and Related Diseases in Indonesia.
Bruni, L., Albero G., Serrano B., Mena M., Gomez D., Munoz J., Bosch FX, de Sanjose S. ICO/IARC Information Centre
on HPV and Cancer (HPV Information Centre). (2019). Human Papillomavirus and Related Diseases in the World.
Karlsen, Frank. Mina Kalantari, Andy Jenkins, Erik Pettersen, Gunnar Kristenser, Ruth Holm, Bo Johansson & Bjorn
Hagmar. (1996). Use of Multiple PCR Primer Sets for Optimal Detection of Human Papillomavirus. Journal of
Clinical Microbiology, 34(9), 2095–2100.
Maori Science Learning Center. (2017). What is PCR?. Retrieved from https://www.sciencelearn.org.nz/resources/2347-
what-is-pcr
National Cancer Institute. (2020). HPV and Cancer. Retrieved from https://www.cancer.gov/about-cancer/causesprevention/risk/infectious-agents/hpv-and-cancer
National Center for Biotechnology Information. (2017). Polymerase Chain Reaction (PCR). Retrieved from https://www.ncbi.nlm.nih.gov/probe/docs/techpcr/
Novianti, T., Juniantito, V., Jusuf, A. A., Arida, E. A., Widia, S., Jusman, A., & Sadikin, M. (2019). Prediksi DNA Primer
Gen PGC- 1α Cecak (Hemidactylus platyurus) dengan Metoda Phylogenetic, Multiple Alignment, dan qPCR.
Indonesian Journal of Biotehnology and Biodiversity, 3(1), 39–47.
Prodia Occupational Health Institute. (2014). Kanker Serviks. Retrieved from https://prodiaohi.co.id/kanker-serviks
R. Markham, Nicholas & Michael Zuker. (2005). DINAmelt web server for nucleic acid melting prediction. Nucleic Acids
Research, Vol 33, Web Server Issue, 577–581.
Research, Z. (2019). Quick -DNA TM Miniprep Plus Kit. Retrieved from www.zymoresearch.com
Safitri, T. A., Dessy Nurul Jannah P., & Henny Saraswati. (2018). Gen L1 HPV 16 dan 18 Sebagai Dasae Dalam Desain
Primer Untuk Deteksi Kanker Leher Rahim Dengan In-House Multiplex PCR. Indonesian Journal of Biotehnology
and Biodiversity. 2, 67–71.
Saraswati, H., Febriana Dwi Wahyuni, & Seprianto. (2019). Desain Primer Secara In Silico untuk Amplifikasi Gen cryIII
dari Bacillus thuringiensis Isolat Lokal. Indonesian Journal of Biotechnology and Biodiversity, 3(1), 33–38.
Schiffman, Mark. Nicolas Wentzensen, Sholom Wacholder, Walter Kinney, Julia C. Gage, Philip E. Castle. (2011). Human
Papillomavirus Testing in the Prevention of Cervical Cancer. JNCI Oxford Journals, 103(5), 368–383.
Sasmito, Dinda Eling K. Rahadian Kurniawan, Izzati Muhimmah. (2014). Karakteristik Primer pada Polymerase Chain
Reaction (PCR) untuk Sekuensing DNA. Seminar Nasional Informatika Medis (SNIMed) V.
V. Bouvard, Baan R., Straif K, Erickson BK, Alvarez RD, Huh WK, Am J Obstet Gynecol. (2012). HPV classification and
Indications and follow-up for HPV testing. Retrieved from
Wulandari, Dwi & T. Mirawati Sudiro. (2014). Pengembangan Antivirus Human Papillomavirus Berbasis Molekul Kecil.
MKA, 37(1).
Zuker, Michael. (2003). Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Research, 31(13), 3406–3415.
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