Application of the hybridization AT-tailing method for detection of human immunodeficiency virus RNA in cells and simian immunodeficiency virus RNA in formalin-fixed and paraffin-embedded tissues

An in situ hybridization (ISH) AT-tailing method (HybrAT) was developed for the detection of viral genomes in infected cells and tissues. The method c onsists of hybridization with oligonucleotide probe which has a 3' end olig o d(A-T) tag, followed by elongation of the oligo d(A-T) by Delta Tth DNA p olymerase in the presence of the labeled nucleotide. The in situ HybrAT det ected human immunodeficiency virus type 1 (HIV-1) in cells and simian immun odeficiency virus (SIV) in formalin-fixed and paraffin embedded sections wi th a sensitivity comparable to RNA ISH. The advantage of this method over o ther methods is discussed. (C) 1999 Elsevier Science B.V. All rights reserv ed.