Detection of hepatitis B virus transcriptional activity: an hypothesis.
This paper outlines a method of demonstrating hepatitis B virus (HBV)-related transcriptional activity based on the random amplification of cDNA ends (RACE) protocol. Following RNA extraction from the appropriate tissue, cDNA is synthesized using a long oligo d(T) based primer containing unique 5' oligonucleotide sequences. Nested polymerase chain reaction (PCR) is then carried out using virus-specific oligonucleotides and standard PCR (approximately 20 base pairs) oligonucleotides homologous to 5' regions of the long oligo-d(T) primer used to initiate the cDNA reaction. This technique is particularly applicable to the study of HBV transcription in small samples of tissue, for example, from renal biopsies obtained from patients with HBV-related nephritis.
Sallie
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Liver Diseases Section, NIDDK, National Institutes of Health, Bethesda, Maryland 20892, USA.
D001482:Base Composition; D001483:Base Sequence; D017931:DNA Primers; D018076:DNA, Complementary; D006509:Hepatitis B / Q000150:complications; D006515:Hepatitis B virus / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000821:virology; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009393:Nephritis / Q000821:virology; D016133:Polymerase Chain Reaction; D019105:Random Amplified Polymorphic DNA Technique; D014158:Transcription, Genetic
D017931:DNA Primers; D018076:DNA, Complementary