Detection of Non-Coding RNA (Hsa_circ_0003416) in Pulmonary Arterial Hypertension Patients’ Plasma by qRT-PCR Analysis

Fajri Marindra Siregar, Dicka Wahyu Setiasari, Anggoro Budi Hartopo, Lucia Kris Dinarti, Sofia Mubarika Haryana

2023

Abstract

Circular RNA refers to a type of non-coding RNA molecule and possesses a circular conformation. Circular RNA has greater stability compared to linear RNA due to its resistance to RNAse activity. hsa_circ_0003416 is a circular RNA reported to have potential as a biomarker in patients with pulmonary arterial hypertension (PAH). This study aims to detect hsa_circ_0003416 using qRT-PCR in the plasma of PAH patients. The sample was taken from RSUP Dr. Sardjito Yogyakarta. The miRNeasy Serum/Plasma Advanced kit was used to isolate total RNA. Following that, cDNA was generated with an Applied Biosystems thermal cycler and the ExcelRTTM Reverse Transcription Kit II reagent, and qPCR was performed with an Applied BiosystemsTM 7500 Real-Time PCR equipment and the SensiFASTTM SYBR® Lo-ROX Kit reagent. Specific divergent primers were utilized. Melting curve analysis and visualization of the amplified products were performed using gel electrophoresis. The qRT-PCR technique achieved a single amplification, resulting in a melting curve value observed at 81.8oC. Examination by visualizing the gel electrophoresis results showed a single band that matched the target size, specifically 124 base pairs. In conclusion, the qRT-PCR analysis successfully identified the presence of hsa_circ_0003416 in the plasma of PAH patients.

Paper Citation

in Harvard Style

Siregar F., Setiasari D., Hartopo A., Dinarti L. and Haryana S. (2023). Detection of Non-Coding RNA (Hsa_circ_0003416) in Pulmonary Arterial Hypertension Patients’ Plasma by qRT-PCR Analysis. In Proceedings of the 1st International Conference on Medical Science and Health - Volume 1: ICOMESH; ISBN 978-989-758-740-5, SciTePress, pages 335-339. DOI: 10.5220/0013672000003873

in Bibtex Style

@conference{icomesh23,
author={Fajri Marindra Siregar and Dicka Wahyu Setiasari and Anggoro Budi Hartopo and Lucia Kris Dinarti and Sofia Mubarika Haryana},
title={Detection of Non-Coding RNA (Hsa_circ_0003416) in Pulmonary Arterial Hypertension Patients’ Plasma by qRT-PCR Analysis},
booktitle={Proceedings of the 1st International Conference on Medical Science and Health - Volume 1: ICOMESH},
year={2023},
pages={335-339},
publisher={SciTePress},
organization={INSTICC},
doi={10.5220/0013672000003873},
isbn={978-989-758-740-5},
}

in EndNote Style

TY - CONF

JO - Proceedings of the 1st International Conference on Medical Science and Health - Volume 1: ICOMESH
TI - Detection of Non-Coding RNA (Hsa_circ_0003416) in Pulmonary Arterial Hypertension Patients’ Plasma by qRT-PCR Analysis
SN - 978-989-758-740-5
AU - Siregar F.
AU - Setiasari D.
AU - Hartopo A.
AU - Dinarti L.
AU - Haryana S.
PY - 2023
SP - 335
EP - 339
DO - 10.5220/0013672000003873
PB - SciTePress