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, characteriz ed by hyperspasticity and ataxia. We have used PCR to amplify, clone a nd 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 sequences established that all HTLV-IIA sequences contained a unique H inf 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 SK11O/SK111 and subsequent digestion with these enzymes, C oncordance between sequenced and OR-based subtyping of DNA amplified b y PCR was absolute among 22 HTLV-II isolates tested. Further OR or seq uence analyses on an additional 30 other isolates indicated that the m ajority 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 fo rmat should facilitate molecular epidemiology studies of HTLV-II infec tion and allow for subtype stratification in assessing the sensitivity and specificity of HTLV detection formats and HTLV-II disease associa tion. (C) 1995 Wiley-Liss, Inc.