Detection and quantitation of subgroup C adenovirus DNA in human tissue samples by real-time PCR.
Advances in amplification techniques have revolutionized the ability to detect viruses both quantitatively and qualitatively and to study viral load. Real-time polymerase chain reaction (PCR) amplification depends on the ability to detect and quantify a fluorescent reporter molecule whose signal increases in proportion to the amount of amplification product generated. Recent advances have been made by using probes, such as TaqMan probes, to detect amplified products. Use of these probes offers confirmation of specificity of the PCR product. Here we describe a sensitive real-time PCR assay to quantify subgroup C adenoviral DNA in human lymphocytes derived from mucosal tissues removed in routine tonsillectomy or adenoidectomy. This chapter will describe in detail the methods used for these analyses.
Garnett
C T
CT
Laboratory of Tumor Immunology and Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
Pao
Ching-I
CI
Gooding
Linda R
LR
D000256:Adenoviridae / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics / Q000302:isolation & purification; D017931:DNA Primers; D004261:DNA Replication; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008214:Lymphocytes / Q000821:virology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*
D036022:Capsid Proteins; D017931:DNA Primers; D004279:DNA, Viral; C018315:hexon capsid protein, Adenovirus