Document 2749
 DOCN  M94A2749
 TI    Simultaneous analysis of retroviruses by PCR and cohybridization.
 DT    9412
 AU    Di Macco E; Aebischer ML; Angeloni U; Civita G; Giuliacci S; Matarazzo
       P; Angeloni P; Central Laboratory, Italian Red Cross, Rome.
 SO    Int Conf AIDS. 1994 Aug 7-12;10(1):233 (abstract no. PB0361). Unique
       Identifier : AIDSLINE ICA10/94369826
 AB    OBJECTIVES. A coamplification and cohybridization protocol was
       established to evaluate the simultaneous detection of HIV 1, HTLV 1 and
       HTLV 2 specific sequences using a non isotopic method. METHODS. Genomic
       DNA was extracted from mononuclear cells of infected individuals by
       Proteinase K treatment. About 2 micrograms of genomic DNA were subjected
       to PCR test using SK 38-39 primers for HIV 1 detection (GAG gene) and
       M1259-A6 primers for HTLV 1/2 detection (TAX-REX gene). PCR was
       performed in one tube using four primers. Amplified products were
       hybridized with specific probes and revealed by DEIA technique (Sorin
       Biomedica, Saluggia Italy). Probe mixtures were dispensed in each well
       in various ratios. Positive DNA for HIV 1 or HTLV 1 or HTLV 2 viruses
       were also analyzed in single PCR and separate wells. RESULTS. PCR
       analysis and hybridization resulted extremely specific to detect each
       virus. The absorbances obtained with positive PCR products did not
       reveal any difference using one or three specific probes. CONCLUSIONS.
       The application of this protocol to pools of blood donors mononuclear
       cells would allow the screening of HIV 1, HTLV 1 and HTLV 2 viruses by
       PCR with a low cost.
 DE    Base Sequence  DNA Probes/DIAGNOSTIC USE  DNA, Viral/GENETICS/ISOLATION
       & PURIF  Human  HIV Infections/*DIAGNOSIS/MICROBIOLOGY  HIV-1/*GENETICS
       HTLV-I/*GENETICS  HTLV-I Infections/*DIAGNOSIS/MICROBIOLOGY
       HTLV-II/*GENETICS  HTLV-II Infections/*DIAGNOSIS/MICROBIOLOGY  *Nucleic
       Acid Hybridization  Polymerase Chain Reaction/*METHODS  MEETING ABSTRACT

       SOURCE: National Library of Medicine.  NOTICE: This material may be
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