Hepatitis E Virus (HEV) is the causative agent of an acute and self-limited form of hepatitis. The virus is transmitted by the faecal–oral route and is a major cause of viral hepatitis in much of the developing world where it causes sporadic infections and large-scale epidemics. A simple and rapid protocol for the measurement of HEV faecal shedding by a real-time polymerase chain reaction (PCR) with the SYBR Green method on a LightCycler instrument, is described. After only 3 h the real-time quantitative PCR method detected 10 molecules of HEV cDNA fragment per reaction tube and showed a high linear dynamic range of quantitation (10–106 molecules of cDNA/reaction) with a good correlation (r = −1.00). Its specificity was confirmed by assay in human faecal samples. © 2004 Elsevier B.V. All rights reserved.
Detection and quantitation of hepatitis E virus in human faeces by real-time quantitative PCR
ORRU, GERMANO;MASIA, GIUSEPPINA;PIRAS, VINCENZO;COPPOLA, ROSA
2004-01-01
Abstract
Hepatitis E Virus (HEV) is the causative agent of an acute and self-limited form of hepatitis. The virus is transmitted by the faecal–oral route and is a major cause of viral hepatitis in much of the developing world where it causes sporadic infections and large-scale epidemics. A simple and rapid protocol for the measurement of HEV faecal shedding by a real-time polymerase chain reaction (PCR) with the SYBR Green method on a LightCycler instrument, is described. After only 3 h the real-time quantitative PCR method detected 10 molecules of HEV cDNA fragment per reaction tube and showed a high linear dynamic range of quantitation (10–106 molecules of cDNA/reaction) with a good correlation (r = −1.00). Its specificity was confirmed by assay in human faecal samples. © 2004 Elsevier B.V. All rights reserved.
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