Optimasi Primer : Suhu Annealing untuk Deteksi Gen Matrix Virus Influenza A Menggunakan  Polymerase Chain Reaction (PCR)

Authors

DOI:

https://doi.org/10.30605/e9cf4f40

Keywords:

Optimization, Polymerase Chain Reaction, Annealing temperature, Primers, Avian Influenza Viruses

Abstract

Influenza A virus (IAV) is a pathogen that attacks the respiratory tract in humans and animals, which can be a threat to public health. This virus has a number of genes that are targeted by PCR for diagnostic detection purposes, one of which is the matrix gene. Detection of the matrix gene in the Influenza A virus from DNA/RNA genetic material can be done using the polymerase chain reaction (PCR) technique. One of the stages that determines the success of the PCR process is the annealing stage, for which the optimum temperature and cycle are required for primer attachment in the annealing stage. This research is a descriptive study towards an experiment conducted in June 2025 at the Bacteriology Laboratory of the Research and Innovation Agency (BRIN) KST. Soekarno Cibinong. The results obtained showed that the optimum temperature for matrix gene amplification in the Influenza A virus using the forward primer FluAV M-U44 and reverse primer FluAV M-L287 was 53.5 °C, 54 °C, and 54,5 °C and 35 cycles were evidenced by the formation of a fairly bright band in the electrophoresis results.

References

Abhishek, V., Gopalareddy, S., & Priya Eswaran, S. (2025). Prevalence of Influenza Viruses A and B in Seasonal Flu : A Tertiary Care Hospital Study. International Journal of Current Microbiology and Applied Sciences, 14(3), 170–179. https://doi.org/10.20546/ijcmas.2025.1403.020

Anggreni, D., Wahyuni, S., & Suwandi, I. (2024). Optimasi suhu annealing PCR untuk deteksi gen spesifik pada virus unggas. Jurnal Bioteknologi Indonesia, 19(2), 85-92.

Anthony SJ, et al. 2012. Emergence of fatal avian influenza in New England harborseals. MBio. 3(4)

Atifa, Y., Rahman, M. A., & Dewi, F. R. (2023). Penerapan variasi suhu annealing dalam optimasi PCR terhadap spesifisitas hasil amplifikasi. Jurnal Bioteknologi dan Biomolekuler, 18(3), 121-130.

Chen, X., et al. (2022). PCR optimization strategies for viral RNA detection. Molecular Biotechnology, 64(5), 528-540.

Fira Safitri dan Afifatul Achyar. 2023. In Silico Primer Design For Chicken ND5 Gene Amplification By Real Time Polymerase Chain Reaction (rtPCR). Serambi biologi. Vol. 8 No. 3

Frisa, A., Elfidasari, D., & Murtini, S. (2017). Seroprevelensi Virus Avian Influenza SubTipe H5N1. 4(2), 74–82.

Gladys, S., Putri, N. A., & Hidayati, R. (2023). Pengaruh suhu annealing terhadap amplifikasi PCR: Studi kasus pada gen bakteri patogen. Indonesian Journal of Biotechnology, 17(1), 45-53.

Lee, S. H., et al. (2020). Effects of annealing temperature on PCR specificity and efficiency. Biochemical and Biophysical Research Communications, 531(1), 280-285.

Mackay, I. M., et al. (2017). Real-time PCR in virology. Journal of Clinical Virology, 104, 87-94.

Pertiwi, E. A., Hasanah, S., & Firdaus, F. (2015). Optimasi suhu annealing PCR untuk amplifikasi gen 16S rRNA bakteri. Jurnal Ilmu Farmasi, 12(1), 25-31.

Purwanto, B., Nurrahman, H. H., & Wijaya, A. F. (2018). Optimasi metode PCR untuk deteksi patogen tanaman dengan variasi suhu annealing. Jurnal Bioteknologi, 15(2), 88-94.

Putri, D. H., Rahman, A. A., & Sari, M. N. (2021). Optimasi suhu annealing PCR untuk amplifikasi gen target pada virus. Jurnal Biologi Terapan, 9(2), 89-96.

Sari, D. K., et al. (2021). Detection of avian influenza virus in domestic poultry using optimized PCR protocols. Veterinary Research Communications, 45(3), 159-168.

Shtykova, E. V, Baratova, L. A., Fedorova, N. V, Radyukhin, V. A., Ksenofontov, A. L., Volkov, V. V, Shishkov, A. V, Dolgov, A. A., Shilova, L. A., Batishchev, O. V, Jeffries, C. M., & Svergun, D. I. (2013). Structural Analysis of Influenza A Virus Matrix Protein M1 and Its Self-Assemblies at Low pH. 8(12), 1–10. https://doi.org/10.1371/journal.pone.0082431

Sisca, M., Prasetyo, E., & Wulandari, A. (2022). Peran siklus amplifikasi pada efektivitas PCR untuk deteksi patogen. Jurnal Mikrobiologi Indonesia, 15(1), 45-52.

Sundararajan, V., Kumar, P. S., & Rajasekaran, K. (2021). Suhu annealing dalam PCR dan pengaruhnya terhadap spesifisitas amplifikasi. Journal of Molecular Diagnostics, 23(4), 510-517.

Wijayanti, N., & Permadi, D. (2022). Sensitivity increase in avian influenza detection using optimized annealing temperature. Journal of Molecular Diagnostics, 24(2), 241-250

Zhang, Y., et al. (2018). Optimization of PCR annealing temperature for sensitive viral detection. Applied Microbiology and Biotechnology, 102, 8639-8648.

Downloads

Published

2026-04-20

Issue

Vol. 11 No. 2 (2026): April - Juni 2026

Section

Articles

License

Copyright (c) 2026 Natalia Turnip, Nova Dilla Yanthi, Dwi Hilda Putri

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

In submitting the manuscript to the journal, the authors certify that:

  • They are authorized by their co-authors to enter into these arrangements.
  • The work described has not been formally published before, except in the form of an abstract or as part of a published lecture, review, thesis, or overlay journal.
  • That it is not under consideration for publication elsewhere,
  • That its publication has been approved by all the author(s) and by the responsible authorities – tacitly or explicitly – of the institutes where the work has been carried out.
  • They secure the right to reproduce any material that has already been published or copyrighted elsewhere.
  • They agree to the following license and copyright agreement.

License and Copyright Agreement

Authors who publish with this journal agree to the following terms:

  1. Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under Creative Commons Attribution License (CC BY 4.0) that allows others to share the work with an acknowledgment of the work's authorship and initial publication in this journal.
  2. Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in this journal.
  3. Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.

How to Cite

Optimasi Primer : Suhu Annealing untuk Deteksi Gen Matrix Virus Influenza A Menggunakan  Polymerase Chain Reaction (PCR) . (2026). Jurnal Biogenerasi, 11(2), 510-515. https://doi.org/10.30605/e9cf4f40