A rapid and sensitive method of identification of HTLV‐II subtypes

Abstract
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 sequences 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.