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Journal : indonesian journal of medical laboratory science and technology

Efficiency and challenges of exosome and microRNA extraction from clinical biofluids in Indonesia: A commercial kit-based approach for qPCR applications Aprilia Indra Kartika; Muchamad Dafip; Nastiti Wijayanti; Didik Setyo Heriyanto; Sofia Mubarika Haryana; Kartika Widayati Taroeno-Hariadi
JURNAL INDONESIA DARI ILMU LABORATORIUM MEDIS DAN TEKNOLOGI Vol 8 No 2 (2026): In Progress
Publisher : Universitas Nahdlatul Ulama Surabaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.33086/ijmlst.v8i2.7350

Abstract

Exosome miRNA has significant potential as biomarker for minimal or non-invasive diagnosis and monitoring of disease progression. However, extraction of miRNA exosomes from biofluids is challenging, and there is a lack of comparative evaluation of commercially available extraction kits for qPCR applications. Therefore, this study aims to obtain information about miRNA exosome extraction using commercial kits and analyze its expression with the qRT-PCR method. The exosome extraction was carried out using 3 types of commercial kits, while the RNA extraction was performed with a spin column. The purity and concentration of exosomes were measured using NTA, while total RNA was measured with nanodrops. Subsequently, miRNA expression was measured using qRT-PCR. The results showed that the concentration of exosomes obtained using kit 1 for plasma isolation was 3.5 x 1011 with particle sizes ranging from 30-240 nm. Kit 2 yielded a value of 2.9 x 1010 with particle sizes ranging from 30-150 nm, while kit 3 had no measurable value. The average concentration of RNA extracted using the RNA Basic kit from HansaBioMed was 42.53 ng/μL, while the exoRNeasy Midi kit from Qiagen yielded 45.28 ng/μL. Template RNA of 100 ng was converted into cDNA, while the cDNA was diluted at a ratio of 1:20 using RNase free water during the quantification process of miRNA exosome expression with the qRT-PCR method. Based on the results, exosome miRNA quantification process requires the use of the correct method at each stage to obtain the right expression for clinical studies.
Co-Authors . Sismindari Abdul Rahman Siregar, Abdul Rahman Abrory Agus Cahya Pramana Adelfiani, Adelfiani Adi Sofyan Ansori, Muhammad Aeniah, Siti Agustiningsih Agustiningsih Akira Hosoyama Allimi, Hayu Swari Amalia, Nur Rofika Ayu Shinta Andi Mahendra Andrian, Krisna Noli Anika Prastyowati, Anika Aprilia Indra Kartika Apriliyani, Tia Ardaning Nuriliani Ari Susilowati Ari Susilowati Ariesti, Wiwin Arif Luthfi Nurul Huda Aris Haryanto Atsushi Yamazoe Aushia Tanzih Al Haq Bambang Hariwiyanto Bambang Retnoaji BUDI SAKSONO Budi Saksono Camelia Herdini Ch. Retna Handayani Dafa, Maula Haqul Dafip, Muchamad Dewi Kartikawati Paramita Diani Mentari Didik Setyo Heriyanto Dwi Liliek Kusindarta Dwi Liliek Kusindarta Dyah Widiastuti Empra, Desi Eka Putri ERLIN LISTIYANINGSIH Fajar Sofyantoro Hakim, Mohamad Saifudin Halim, Shuha Ma’muriyah Hanbal, Mayland Muhammad Hendry Tri Sakti Saragih Hera Nirwati Heriyanto, Didik S. Hevi Wihadmadyatami Hideaki Nojiri Ibnu Agus Ariyanto Illiyin, Sirayya Ina Kusrini Jajah Fachiroh, Jajah JAKA WIDADA Juliadmi, Dian Karlina, Ina Karnati, Srikanth Kartika W. Taroeno-Hariadi Kartika Widayati Taroeno-Hariadi Kartika, Aprilia I. Lisna Hidayati Mayani, Olvita Mei Ria Santi Michael Kann Muchamad Dafip Muhammad Adnan Mumtazah, Dzul Fithria Munawir Sazali Murwantoko . Naima, Mirtani Nenny Sri Mulyani Ni Wayan Erly Sintya Dewi Nisa, Mutia Khoirun Nuhamunada, Matin Oktaviana, Shintia Peni Indrayudha Pertiwi, Anggita Endar Pertiwi, Gian Aditya Podhi, Felisitas Moli Pradnya Paramita Pratitis, Visi Endah Primahesa, Alfian Prissandi, Anthera Al Firdaus Priyambada, Fajar Purnomo Hadi Pusparini, Nur Ainun Oktavia Puspitasari, Pinki Anggrahini Putra, Hendy Eka R.C. Hidayat Soesilohadi Rahadian Yudo Hartantyo Rahayu, Sekar Ramadhani, Eka Rarastoeti Pratiwi Retno Murwanti Rinda Binugraheni, Rinda Rohmah, Zuliyati Rumansara, Papuani Samparisna Sadewo, Imran Saeed, Faisal Salamah, Rohmi Salsabila Faradisa Nuris Saragih, Hendry T. S. S. G. Sari, Ria Vinola Septhya Saribu, Ruth Liananda Citra Dolok Septriani, Nur Indah Septriani, Nur Indah Setiawibawa, Raden Aditya Aryandi Shinta Hartanto Simanungkalit, Eben Ezer Slamet Widiyanto Slamet Widiyanto Sofia M. Haryana Sofia Mubarika Haryana Sofia Mubarika Haryana Subchan, Aditya Nur Sularsah, Sais Supraitno, Murtiadi Erlan Sutaryo1 S, Sutaryo1 T. Trijoko Tasania, Nadia Nisa Titis Widowati, Titis To'bungan, Nelsiani Tri Rini Nuringtyas Tri Wibawa Triska Desi Sundari Tsabitah, Khansa Wiwin Ariesti Wulansari, Riska Yundari, Yundari Zulfa, Amania Zulfadhli Zulfadhli Zumrotus Sholichah Zusrina, Laili Mufli ‘Aliyah, Siti Hamidatul