Recombinase polymerase amplification as a promising tool in hepatitis C virus diagnosis

Abstract

Hepatitis C virus (HCV) infection represents a significant health problem and represents a heavy load on some countries like Egypt in which about 20% of the total population are infected. Initial infection is usually asymptomatic and result in chronic hepatitis that give rise to complications including cirrhosis and hepatocellular carcinoma. The management of HCV infection should not only be focus on therapy, but also to screen carrier individuals in order to prevent transmission. In the present, molecular detection and quantification of HCV genome by real time polymerase chain reaction (PCR) represent the gold standard in HCV diagnosis and plays a crucial role in the management of therapeutic regimens. However, real time PCR is a complicated approach and of limited distribution. On the other hand, isothermal DNA amplification techniques have been developed and offer molecular diagnosis of infectious dieses at point-of-care. In this review we discuss recombinase polymerase amplification technique and illustrate its diagnostic value over both PCR and other isothermal amplification techniques.

Nucleic Acid Amplification using Recombinase Polymerase: Enzymatic Approach

“…RPA is an isothermal (heat independence) nucleic acid amplification technique [5]. Using a combination of 3 enzymes the amplification will be achieved within 20 minutes.…”

Rapid Detection of Cyprinid Herpesvirus 3 in Latently Infected Koi by Recombinase Polymerase Amplification

“…Recombinase polymerase amplification (RPA) is an emerging method for the isothermal amplification of nucleic acid (Piepenburg et al 2006), and has been employed for the detection of various pathogens (Euler et al 2012a, 2012b; Abd El Wahed et al 2013a, 2013b; Amer et al 2013; Boyle et al 2013, 2014; Ahmed et al 2014; del Río et al 2014; Jaroenram and Owens 2014; Krõlov et al 2014; Mekuria et al 2014; Tsaloglou et al 2015; Xia et al 2014, 2015; Zaghloul and El-Shahat 2014; Zhang et al 2014). The RPA method utilizes three main enzymes to amplify template DNA: recombinases, single stranded binding protein (SSB), and a strand-displacing polymerase (Piepenburg et al 2006).…”

Evolución de técnicas de diagnóstico de virus fitopatógenos

“…com use LAMP technology, to mention a few. All these methodologies have advantages and disadvantages, according to the compilation made by Zaghloul, H and El-shahat, M (2014); the only one of these technologies that only has advantages is RPA.…”