PMID: 9643505Jun 27, 1998Paper

Hepatitis C genotyping by direct sequencing of the product from the Roche AMPLICOR test: methodology and application to a South Australian population

Pathology
J HollandG D Higgins

Abstract

The Roche AMPLICOR RT-PCR amplifies a 244 nucleotide sequence within the 5' non coding region (5'NCR) of the viral genome and is a widely used commercial test for the qualitative determination of hepatitis C RNA from sera. This paper describes a routine procedure for the purification of the PCR product, and its use in automated DNA sequencing, for determining the genotype of hepatitis C virus (HCV) isolates. Direct sequencing of the purified product was possible for 86% of samples, whilst 14% required additional amplification using a nested PCR method in order to read the resulting electropherogram. This method of genotyping is considerably less expensive than currently available commercial kits, and is convenient for the increasing number of laboratories that have access to automated DNA sequencers. The highly conserved nature of the 5'NCR limited differentiation of types and subtypes to an extent comparable to commercial HCV typing methods. Using this method on available laboratory samples and on patients about to commence interferon therapy, we found a predominance of genotype 1 (59%) and 3a (31%). Analysis of data on the interferon patients showed the median length of time from first exposure to diagnosis to be significantl...Continue Reading

References

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Citations

May 22, 2003·Journal of Virological Methods·Laura RossiEnrico M Silini
Apr 28, 2001·Journal of Clinical Microbiology·P HalfonD Ouzan
Feb 4, 2006·Journal of Clinical Microbiology·Jeffrey J GermerJoseph D C Yao
Sep 30, 2009·Clinical Biochemistry·Hongju MaoYongzhong Wang
Nov 3, 2007·Clinica Chimica Acta; International Journal of Clinical Chemistry·Hongju MaoMengsu Yang
Oct 27, 2005·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Regino P González-PeraltaUNKNOWN International Pediatric Hepatitis C Therapy Group
Sep 15, 2004·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Yi-Wei TangBelinda Yen-Lieberman
Sep 25, 2016·Archives of Virology·Hosam ZaghloulHossam El-Sharawy
Jan 11, 2005·World Journal of Gastroenterology : WJG·Motokazu MukaideMasashi Mizokami
Aug 8, 2013·Hepatitis Monthly·Keivan Majidzadeh-AHossain Jabbari
Oct 20, 2018·PloS One·María de la Luz Martínez-RodríguezAlicia Ocaña-Mondragón
Aug 23, 2008·Revista Da Sociedade Brasileira De Medicina Tropical·Carlos PeroneRosângela Teixeira
Jan 9, 2014·Indian Journal of Medical Microbiology·M RastinH R Panah
Feb 3, 2000·Journal of Clinical Microbiology·P A WhiteW D Rawlinson
Jun 29, 1999·Analytical Chemistry·C L Mouritsen, D R Hillyard

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