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All Journal Microbiology Indonesia Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Medical Journal of Indonesia Dental Journal (Majalah Kedokteran Gigi) Universa Medicina Indonesian Journal of Obstetrics and Gynecology (Majalah Obstetri dan Ginekologi Indonesia) Eksakta : Berkala Ilmiah Bidang MIPA Jurnal Biomedika dan Kesehatan
Yeva Rosana
Department Of Microbiology, Faculty Of Medicine, Universitas Indonesia, Jakarta 10430
Agriculture, Biological Sciences & Forestry Astronomy Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Civil Engineering, Building, Construction & Architecture Computer Science & IT Dentistry Energy Engineering Environmental Science Health Professions Immunology & microbiology Materials Science & Nanotechnology Mathematics Mechanical Engineering Medicine & Pharmacology Neuroscience Public Health

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Journal : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Development of a SYBR Green real-time PCR-based assay system for detection of Neisseria gonorrhoeae Andi Yasmon; Rela Febriani; Louisa Ivana Utami; Fithriyah Fithriyah; Yeva Rosana; Fera Ibrahim; Pratiwi Sudarmono
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 54, No 1 (2022)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.19106/JMedSci005401202201

Abstract

Diagnosis of Neisseria gonorrhoeae infection is needed for patient therapy and for reducing this bacterial transmission in the population. The culture method is a gold standard method for N. gonorrhoeae detection, however it has low sensitivity. Among molecular methods with high sensitivity and specificity, SYBR Green real-time PCR is the potential method for N. gonorrhoeae detection. In this study, we developed an SYBR Green real-time PCR-based system assay for N. gonorrhoeae detection. Several PCR conditions were optimized and analyzed including primer annealing temperature, DNA template volume, the limit of detection (LoD), cross-reaction with others (bacteria, viruses, fungus, protozoa), and quality assurance. The results showed that the annealing temperature and DNA template volume were 60oC and 5 µL, respectively. The LoD was 29 DNA copies corresponding to 3 bacterial cells per reaction. No cross-reaction was detected for other bacteria, viruses, fungus and protozoa. The external quality assurances enrolled in 2019 and 2021 showed 100% concordance. The preliminary testing for clinical samples was also 100% concordance. In conclusion, the SYBR Green real-time PCR-based system assay developed in this study is promising for application in clinical laboratories.
Co-Authors Abas, Ghina Mutiara Andi Yasmon Anggraini Suwarsono, Erike Anis Karuniawati Ariyani Kiranasari Bachti Alisjahbana Bachti Alisyahbana Basri A. Gani Beti E. Dewi Conny R. Tjampakasari Conny Tjampakasari Dick van Soolingen Dwiana Ocviyanti E. Risdiyani Efendi, Ida Effendi, Ida Fera Ibrahim Ferry Darmawan Fithriyah Fithriyah Gultom, Desy A. Gus Permana Subita Herliyana, Lina Ibnu Agus Ariyanto Ida Parwati Ika Ningsih Imaniar, Clara Krisandi, Grady Louisa Ivana Utami Mardiastuti H Wahid Pratiwi Pujilestari Sudarmono Prawoto Prawoto Rahmah Amran Reinout van Crevel Rela Febriani Retno Kadarsih S. Nilawati S. Sunnati Shanty Olivia Siti Aliyah Pradono SJATHA, FITHRIYAH Sri Rezeki SYADZA RHIZKY PUTRI AKHMAD T. M. Sudiro TETSUHIRO MATSUZAWA Tjahjani M. Sudiro TOHRU GONOI Wia Melia Wresti Indriatmi B. Makes