A Novel and Simple Method for HCV Genotyping

Document Type : Research Article

Authors

• Ali Nazemi ¹
• Elham Sadeghi Tazehabadi ²
• Mostafa Jafarpoor ²
• Sahrashoob Sharifi ³

¹ Department of Genetics, Islamic Azad University-Tonekabon branch, Mazandaran, Iran

² Department of Microbiology, Islamic Azad University-Tonekabon branch, Mazandaran, Iran

³ Department of Medical Biology, Ankara University, Ankara, Turkey

Abstract

Hepatitis C virus (HCV) is a major global health problem and one of the most common causes of chronic liver diseases. The current study was carried out to investigate the ability of High Resolution Melting (HRM) method in HCV genotyping. 40 HCV-Positive sera with unknown and 12 sera with known HCV genotypes were collected from different clinical laboratories in Tehran as well as Rasht in north of Iran. The RT-PCR was performed on RNA extracted from each sample. The first PCR amplification product was used as template for second run in a Nested-PCR followed by HRM method. Unknown genotypes were determined with comparing of their normalized and difference graphs with those obtained by known genotypes as standard. All results were confirmed by direct sequencing. In a comparison between results obtained by HRM method and those of direct sequencing, only 2 out of 40 samples with unclear HCV genotypes had been incorrectly genotyped by HRM method. The results obtained by this study show that HRM can be introduced as a simple, fast, and reliable method for HCV genotyping in clinical laboratories.

Keywords

• hepatitis C virus
• Genotyping
• High Resolution Melting

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