Objective and Design:
To develop a method for the detection of human T-cell lymphotropic virus type I (HTLV-I) in archival biopsy specimens. A polymerase chain reaction—based gene amplification method was developed to detect HTLV-I proviral DNA in paraffinembedded specimens. The specificity of the polymerase chain reaction products was controlled by Southern blot analysis using a nested oligonucleotide probe and by nucleotide sequencing. The nucleophosmin gene and the T-cell receptor-γ gene were used as controls for the integrity and adequacy of total DNA and T-cell DNA, respectively. This study was conducted with patients referred to an academic medical center. Biopsy specimens were obtained from lesional skin or lymph node from Japanese patients with HTLV-I seropositive adult T-cell leukemia/lymphoma. The main outcome measure was the ability to detect HTLV-I pX region proviral DNA.
Comparative analysis of DNA extracted from fresh samples of the HTLV-I infected MT4 T-cell line demonstrated that formalin fixation and paraffin embedding resulted in a 100-fold reduction in sensitivity of the assay. Nevertheless, HTLV-I pX sequences were still readily detectable in paraffin-embedded samples of MT4 T cells and adult T-cell leukemia/lymphoma specimens. Both formalin and B5 fixation were suitable for the assay that was 100% specific for HTLV-I—infected tissues.
The use of this method should greatly facilitate investigation of the role of HTLV-I in human diseases by allowing analysis of a wide variety of archival tissue specimens. In addition, the controls designed for the current study can be used in a variety of other polymerase chain reaction—based studies of T cells to ensure against false-negative results caused by DNA degradation or inadequate T-cell density.Arch Dermatol. 1996;132:1339-1343
Wood GS, Ruffo A, Salvekar A, Henghold W, Takeshita M, Kikuchi M. Detection of Human T-Cell Lymphotrophic Virus Type I in Archival Tissue Specimens. Arch Dermatol. 1996;132(11):1339-1343. doi:10.1001/archderm.1996.03890350081013