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1994-10-25
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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
protected by Copyright Law (Title 17, U.S.Code).