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 consists of hybridization with oligonucleotide probe which has a 3' end oligo d(A-T) tag, followed by elongation of the oligo d(A-T) by deltaTth DNA polymerase in the presence of the labeled nucleotide. The in situ HybrAT detected human immunodeficiency virus type 1 (HIV-1) in cells and simian immunodeficiency virus (SIV) in formalin-fixed and paraffin embedded sections with a sensitivity comparable to RNA ISH. The advantage of this method over other methods is discussed.
Nakajima
N
N
AIDS Research Center and Department of Pathology, National Institute of Infectious Diseases, Tokyo, Japan.
Sata
T
T
Hanaki
K
K
Kurata
T
T
Yoshikura
H
H
D000818:Animals; D002460:Cell Line; D005557:Formaldehyde; D015497:HIV-1 / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D008253:Macaca mulatta; D008297:Male; D009838:Oligodeoxyribonucleotides / Q000235:genetics*; D015345:Oligonucleotide Probes / Q000235:genetics*; D016612:Paraffin Embedding; D011067:Poly dA-dT / Q000235:genetics*; D012333:RNA, Messenger / Q000302:isolation & purification; D012367:RNA, Viral / Q000302:isolation & purification*; D011933:Reagent Kits, Diagnostic; D015302:Simian Immunodeficiency Virus / Q000235:genetics* / Q000302:isolation & purification; D016707:Tissue Fixation
D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic; C013630:oligo (dT) poly (A); D005557:Formaldehyde; D011067:Poly dA-dT