Document 0096
 DOCN  M9580096
 TI    Typing human T-cell lymphotropic virus (HTLV-I and HTLV-II) by nested
       polymerase chain reaction: application to clinical specimens.
 DT    9506
 AU    Vallejo A; Garcia-Saiz A; Servicio de Diagnostico y Referencia,
       Instituto de Salud Carlos; III, Madrid, Spain.
 SO    J Virol Methods. 1995 Jan;51(1):9-17. Unique Identifier : AIDSLINE
       MED/95247873
 AB    Human T-cell lymphotropic virus type I and II provirus DNA was detected
       by polymerase chain reaction (PCR). MT-2 (HTLV-I infected), C3/44 Mo
       (HTLV-II infected) cell lines and peripheral blood mononuclear cells
       (PBMNC) from HTLV seropositive samples were used. The procedure consists
       of first amplification which detects both HTLV-I and HTLV-II, and a
       second amplification (nested-PCR) to discriminate between the two
       viruses and to improve sensitivity. Optimal conditions of MgCl2
       concentration and annealing temperature were found for maximal
       amplification and specificity. This method was used for the
       amplification of conserved regions of pol and env genes. 1.5 pg of MT-2
       and 5 pg of C3/44 Mo cell line DNAs were detected using nested-PCR and
       liquid hybridization in the pol system. The env system could detect 1.5
       pg of MT-2 and 1.5 pg of C3/44 Mo cell lines DNAs using nested-PCR and
       liquid hybridization. The pol system can type both HTLV-I and HTLV-II in
       only two steps without the use of type-specific radiolabeled probes.
       Furthermore, this method can detect and discriminate the two viruses in
       one step PCR using the primers used in the nested-PCR. Nevertheless,
       there is a decrease in sensitivity of 100-fold. The results of five
       seropositive samples confirmed by Western blot are compared with PCR.
       PCR typed one of these samples as HTLV-I and the rest as HTLV-II. This
       technique is useful in cases such as window period, perinatal studies
       and when serologic results are not satisfactory.
 DE    Base Sequence  Blotting, Western  Cell Line  Comparative Study  DNA
       Primers/GENETICS  DNA Probes/GENETICS  DNA, Viral/GENETICS  Evaluation
       Studies  Genes, env  Genes, pol  Human
       HTLV-I/*CLASSIFICATION/*GENETICS/ISOLATION & PURIF  HTLV-I
       Infections/DIAGNOSIS/VIROLOGY
       HTLV-II/*CLASSIFICATION/*GENETICS/ISOLATION & PURIF  HTLV-II
       Infections/DIAGNOSIS/VIROLOGY  Molecular Sequence Data  Polymerase Chain
       Reaction/*METHODS/STATISTICS & NUMER DATA
       Proviruses/CLASSIFICATION/GENETICS/ISOLATION & PURIF  Sensitivity and
       Specificity  Support, Non-U.S. Gov't  Virology/METHODS/STATISTICS &
       NUMER DATA  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).