Optimized PCR assay for the detection of TT virus.
A polymerase chain reaction (PCR)-based procedure for the detection of TT virus DNA is described. In this method. total nucleic acid extracted from a small volume of serum or plasma is utilized as a template in PCR employing TT virus specific primers designed to highly conserved regions of the virus genome. Additional sensitivity is obtained by carrying out a second round of amplification. Reactions are analyzed by agarose gel electrophoresis, and samples having an ethidium bromide stainable fragment of the appropriate size in the first and/or second amplification are designated as positive. This protocol allows for the rapid and sensitive detection of TT virus in human plasma or serum.
Leary
T P
TP
Virus Discovery Group, Experimental Biology Research, Abbott Laboratories, North Chicago, IL 60064-6269, USA.
Erker
J C
JC
Chalmers
M L
ML
Desai
S M
SM
Mushahwar
I K
IK
D017124:Conserved Sequence; D017931:DNA Primers; D004266:DNA Virus Infections / Q000175:diagnosis / Q000821:virology; D004267:DNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D004270:DNA, Circular / Q000097:blood / Q000235:genetics; D004279:DNA, Viral / Q000097:blood* / Q000235:genetics; D004996:Ethidium; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013698:Templates, Genetic; D013997:Time Factors
D017931:DNA Primers; D004270:DNA, Circular; D004279:DNA, Viral; D004996:Ethidium