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.
Detection and quantitation of hepatitis E virus in human faeces by real-time quantitative PCR / G. Orrù, G. Masia, G. Orrù, L. Romanò, V. Piras, R.C. Coppola. - In: JOURNAL OF VIROLOGICAL METHODS. - ISSN 0166-0934. - 118:2(2004 Jun 15), pp. 77-82.
Detection and quantitation of hepatitis E virus in human faeces by real-time quantitative PCR
L. Romanò;
2004
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.
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