Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR
Background: Rapid quantification of hepatitis C virus is helpful in determining and monitoring of the disease progression and nature of the virus replication. Objective: The aim of the present study was to establish a fast, specific and sensitive tool for HCV-RNA quantification. Methodology: A t...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Kulliyyah of Medicine, International Islamic University Malaysia
2018
|
| Subjects: | |
| Online Access: | http://irep.iium.edu.my/66273/ http://irep.iium.edu.my/66273/1/MRS2018-32_Poster%20%20Third%20generation%20dye.pdf |
| _version_ | 1848786546691407872 |
|---|---|
| author | M. Saleh Habil, Akrahm Hamzah, Hairul Aini Mustafa, Imad Al-Deen A Talib, Noorlelawati Abdul Rahman, Siti Nurul Fazlin |
| author_facet | M. Saleh Habil, Akrahm Hamzah, Hairul Aini Mustafa, Imad Al-Deen A Talib, Noorlelawati Abdul Rahman, Siti Nurul Fazlin |
| author_sort | M. Saleh Habil, Akrahm |
| building | IIUM Repository |
| collection | Online Access |
| description | Background: Rapid quantification of hepatitis C virus is helpful in determining and monitoring of the disease progression and nature of the virus replication.
Objective: The aim of the present study was to establish a fast, specific and sensitive tool for HCV-RNA quantification.
Methodology: A total of 50 serum samples, comprising of 40 HCV-positive and 10 HCV-negative, were included in our study. RNA was extracted, reverse transcribed, and then subjected to real-time PCR amplification. Real-time PCR using EvaGreen dye and primers targeting a 5’UTR was carried out. Reference samples with known viral load were treated similarly to the unknown samples and used to create the standard curves.
Results: Our method showed a high level of analytical specificity and accuracy, with a low limit of detection (~2 IU/ml). It yielded repeatable results with less than 4% of intra- assay variation. The assay covered a broad dynamic range of quantification, ranging from 0.34 to 6 log IU/ml. The diagnostic sensitivity, specificity, and accuracy were all 100%, indicating neither false positive nor false negative results were obtained.
Conclusion: The developed real time PCR using EvaGreen dye has demonstrated a high analytical and diagnostic performance for HCV quantification, suggesting its potential in clinical diagnosis and management. |
| first_indexed | 2025-11-14T17:10:46Z |
| format | Article |
| id | iium-66273 |
| institution | International Islamic University Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-14T17:10:46Z |
| publishDate | 2018 |
| publisher | Kulliyyah of Medicine, International Islamic University Malaysia |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | iium-662732019-07-12T03:26:25Z http://irep.iium.edu.my/66273/ Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR M. Saleh Habil, Akrahm Hamzah, Hairul Aini Mustafa, Imad Al-Deen A Talib, Noorlelawati Abdul Rahman, Siti Nurul Fazlin QR Microbiology QR355 Virology Background: Rapid quantification of hepatitis C virus is helpful in determining and monitoring of the disease progression and nature of the virus replication. Objective: The aim of the present study was to establish a fast, specific and sensitive tool for HCV-RNA quantification. Methodology: A total of 50 serum samples, comprising of 40 HCV-positive and 10 HCV-negative, were included in our study. RNA was extracted, reverse transcribed, and then subjected to real-time PCR amplification. Real-time PCR using EvaGreen dye and primers targeting a 5’UTR was carried out. Reference samples with known viral load were treated similarly to the unknown samples and used to create the standard curves. Results: Our method showed a high level of analytical specificity and accuracy, with a low limit of detection (~2 IU/ml). It yielded repeatable results with less than 4% of intra- assay variation. The assay covered a broad dynamic range of quantification, ranging from 0.34 to 6 log IU/ml. The diagnostic sensitivity, specificity, and accuracy were all 100%, indicating neither false positive nor false negative results were obtained. Conclusion: The developed real time PCR using EvaGreen dye has demonstrated a high analytical and diagnostic performance for HCV quantification, suggesting its potential in clinical diagnosis and management. Kulliyyah of Medicine, International Islamic University Malaysia 2018-08-29 Article PeerReviewed application/pdf en http://irep.iium.edu.my/66273/1/MRS2018-32_Poster%20%20Third%20generation%20dye.pdf M. Saleh Habil, Akrahm and Hamzah, Hairul Aini and Mustafa, Imad Al-Deen and A Talib, Noorlelawati and Abdul Rahman, Siti Nurul Fazlin (2018) Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR. The International Medical Journal Malaysia, 17 (1). p. 31. E-ISSN 1823-4631 http://iiumedic.net/imjm/v1/download/vol_17_no_3_supp_1/MRS2018-32.pdf |
| spellingShingle | QR Microbiology QR355 Virology M. Saleh Habil, Akrahm Hamzah, Hairul Aini Mustafa, Imad Al-Deen A Talib, Noorlelawati Abdul Rahman, Siti Nurul Fazlin Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR |
| title | Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR |
| title_full | Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR |
| title_fullStr | Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR |
| title_full_unstemmed | Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR |
| title_short | Establishment of simultaneous detection and quantitation of HCV-RNA by third generation intercalating dye real-time PCR |
| title_sort | establishment of simultaneous detection and quantitation of hcv-rna by third generation intercalating dye real-time pcr |
| topic | QR Microbiology QR355 Virology |
| url | http://irep.iium.edu.my/66273/ http://irep.iium.edu.my/66273/ http://irep.iium.edu.my/66273/1/MRS2018-32_Poster%20%20Third%20generation%20dye.pdf |