Comparative performances of enzyme-linked immunosorbent, western blot, and polymerase chain reaction assays for human T-lymphotropic virus type II infection that is endemic among Indians of the Gran Chaco region of South America.
BACKGROUND
Human T-cell lymphoma/leukemia viruses types I and II (HTLV-I and HTLV-II) are related exogenous human retroviruses. The former is definitely pathogenic while disease association with the latter is unclear. There are two subtypes of HTLV-II, A and B. Currently, enzyme-linked immunosorbent assays (ELISAs) based on HTLV-I antigens are used to screen for the presence of HTLV-I and -II antibodies. Confirmation and subtyping are accomplished by Western blot (WB) or ELISAs based on HTLV-I whole viral antigens and/or HTLV-I and HTLV-IIA peptides. The sensitivity and specificity of these serologic assays were compared to those of HTLV-I and-II-specific polymerase chain reaction (PCR) assays in tests on samples from Indians from South America in whom the HTLV-IIB subtype is endemic.


METHODS
Sera from 246 Gran Chaco Indians were evaluated for HTLV antibodies with the use of four ELISAs (Retrotek HTLV-I; Cambridge Biotech rgp21 enhanced HTLV-I/II; Vironostika HTLV-I/II; and Select HTLV-I/II), and a WB assay. Peripheral blood leukocyte DNA from each Indian was analyzed for HTLV-I or HTLV-II pol DNA via PCR. Fifteen of the PCR-positive samples were further subtyped via cloning and sequencing and/or oligomer restriction.


RESULTS
Ninety-seven samples (39%) were positive for HTLV-II by serologic and/or PCR assays. All 15 positive DNA samples that were further analyzed were of the HTLV-IIB subtype and were clustered as a highly conserved phylogenetic group. Comparative analyses indicate that the sensitivity and specificity of the various assays were: PCR, 97 and 100 percent; Retrotek, 70 and 91 percent; Cambridge Biotech, 74 and 96 percent; Vironostika, 73 and 99 percent; Select 72 and 98 percent; and WB, 70 and 100 percent.


CONCLUSIONS
The sensitivities of the tested HTLV serologic assays were comparable. However, the specificity of the Retrotek ELISA was significantly lower than that of the others. When positive, the subtyping assays were very specific. However, PCR assays would seem preferable or to be a necessary adjunct for the sensitive detection of HTLV-IIB infection.
Poiesz
B J
BJ
Department of Medicine, State University of New York, Health Science Center, Syracuse, USA.
Dube
S
S
Jones
B
B
Bryz-Gornia
V
V
Dean
M T
MT
Spicer
T
T
Hengst
J
J
Sayre
K
K
Esteban
E
E
Ferrer
J F
JF
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000595:Amino Acid Sequence; D015153:Blotting, Western; D002648:Child; D002675:Child, Preschool; D004797:Enzyme-Linked Immunosorbent Assay; D005188:False Negative Reactions; D018023:HIV Seronegativity; D006679:HIV Seropositivity / Q000175:diagnosis; D015491:HTLV-II Infections / Q000175:diagnosis / Q000453:epidemiology* / Q000208:ethnology*; D006801:Humans; D007199:Indians, South American; D008875:Middle Aged; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015995:Prevalence