Development of a supersensitive polymerase chain reaction method for human T lymphotropic virus type II (HTLV-II) and detection of HTLV-II proviral DNA from blood donors in Japan.
A supersensitive polymerase chain reaction procedure was developed to detect human T-lymphotropic virus type II (HTLV-II) proviral genome. Six primer pairs covering the various regions of HTLV-II were compared and selected on the basis of specificity and sensitivity. Among them, one primer pair of the pol region of HTLV-II (II pol) was able to amplify and detect even 0.1 fg of the cloned plasmid HTLV-II DNA (seven copies) by regular ethidium bromide staining on polyacrylamide gel. By using this procedure, we screened 189 HTLV-I seropositive blood donors from Yamaguchi and Fukuoka Red Cross Blood Centers, Japan. There were four positive samples detectable with the HTLV-II-specific pol primer pair, as well as with the HTLV-I tax primer pair. The amplified DNAs of two specimens were cloned and sequenced. The sequences of the HTLV-I tax region from both specimens were identical to that of HTLV-I. On the other hand, those of the HTLV-II pol region were identical to that of HTLV-II, except for one base substitution in a clone from one subject. These results indicate that dual infection of HTLV-I and HTLV-II in the same persons occurs among Japanese blood donors.
Hamakado
T
T
Ube Research Laboratory, Fujirebio Inc., Yamaguchi, Japan.
Matsumoto
T
T
Koyanagi
Y
Y
Hayashi
K
K
Fukada
K
K
Kouchiyama
T
T
Yoshida
T
T
Yamamoto
N
N
D001483:Base Sequence; D001782:Blood Donors; D002460:Cell Line; D003001:Cloning, Molecular; D004277:DNA, Single-Stranded / Q000302:isolation & purification; D004279:DNA, Viral / Q000032:analysis*; D015368:Human T-lymphotropic virus 1 / Q000302:isolation & purification; D015367:Human T-lymphotropic virus 2 / Q000302:isolation & purification*; D006801:Humans; D007564:Japan; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011533:Proviruses / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
D004277:DNA, Single-Stranded; D004279:DNA, Viral