A rapid and sensitive method of identification of HTLV-II subtypes.
There are 2 subtypes of human T-cell lymphoma/leukemia virus type II (HTLV-II), A and B. HTLV-II is increasingly associated with rare forms of lymphocytic neoplasia and a neurodegenerative disorder, characterized by hyperspasticity and ataxia. We have used PCR to amplify, clone and sequence 140 bp of the pol gene from many isolates of HTLV-IIA and HTLV-IIB from around the world. Analysis of these and other published sequence established that all HTLV-IIA sequences contained a unique Hinf I site and all HTLV-IIB sequences a unique Mse I site. A rapid and specific oligomer restriction (OR) assay was developed utilizing the primer pair SK110/SK111 and subsequent digestion with these enzymes. Concordance between sequenced and OR-based subtyping of DNA amplified by PCR was absolute among 22 HTLV-II isolates tested. Further OR or sequence analyses on an additional 30 other isolates indicated that the majority of North American non-indian HTLV-II isolates were subtype A, while all Paleo-Amerindian samples, including those from the Seminole of Florida; the Guaymi from Panama; and the Toba, Chorote, Wichi, and Chulupe of Argentina, belonged to subtype B. The SK110/SK111 PCR-OR format should facilitate molecular epidemiology studies of HTLV-II infection and allow for subtype stratification in assessing the sensitivity and specificity of HTLV detection formats and HTLV-II disease association.
Dube
S
S
Department of Medicine and Microbiology/Immunology, SUNY Health Science Center, Syracuse 13210, USA.
Spicer
T
T
Bryz-Gornia
V
V
Jones
B
B
Dean
T
T
Love
J
J
Ferrer
J
J
Esteban
N
N
Harrington
W
W
Glaser
J
J
D001118:Argentina; D001483:Base Sequence; D017931:DNA Primers; D004279:DNA, Viral / Q000097:blood*; D005260:Female; D015751:Genes, pol / Q000235:genetics; D015491:HTLV-II Infections / Q000821:virology*; D015367:Human T-lymphotropic virus 2 / Q000145:classification / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D014481:United States; D014764:Viral Proteins / Q000032:analysis
D017931:DNA Primers; D004279:DNA, Viral; D014764:Viral Proteins