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M94A2701.TXT
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1994-10-25
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Document 2701
DOCN M94A2701
TI Confirmatory tests for HIV detection constructed in Latvia.
DT 9412
AU Svirska R; Konicheva V; Line I; Ose V; Meldrajs J; BMC University of
Latvia, Riga.
SO Int Conf AIDS. 1994 Aug 7-12;10(1):244 (abstract no. PB0404). Unique
Identifier : AIDSLINE ICA10/94369874
AB OBJECTIVE: Blood screening of donors, risk-groups, anonymous persons was
begun in Latvia in 1987. Due to economic difficulties the use of
commercial confirmatory tests is limited, that is why necessity arose to
construct our own ones. METHODS: Sera and blood samples of
HIV-1-positive in ELISA patients were received from National AIDS
centre. Western blotting (WB) and PCR were constructed as confirmatory
tests. Viral material from HIV-1 producing cell lines (Jurkat-tat
III-HIV-1, ACH-2) was used for WB. Primers from gag region of HIV-1 were
used for PCR, DNA was extracted from patients leukocytes, negative and
positive for HIV-1 cell cultures and cells as controls. RESULTS: The
best virus-producing cell cultures were selected for virus production
for WB. In ultracentrifugation procedure we lost the majority of gp
120/160 HIV-1, that's why we used aqueous two-phase systems for
purification of glucosylated antigens. Results were comparable with
those got in commercial Abbott tests. False positive results in ELISA
correlated with positive reaction in WB only for p17--we think some
other process, non-specific for AIDS, can gives such a reaction. PCR
with set of primers from gag gave positive results with DNA from
HIV-positive cell cultures, as well as with DNA's of HIV-1 carriers.
Results were confirmed by hybridisation with specific internal probe
containing corresponding HIV-1 regions. CONCLUSIONS: Home-constructed WB
could be used as confirmatory test in HIV screening. PCR is a valuable
confirmatory test, specially in cases, when WB results are dubious. We
plan to compare virus isolates circulating in Latvia with other known
strains.
DE AIDS Serodiagnosis/METHODS Blotting, Western Comparative Study
*Developing Countries Enzyme-Linked Immunosorbent Assay Human HIV
Seropositivity/*DIAGNOSIS/IMMUNOLOGY HIV-1/*IMMUNOLOGY Latvia
Polymerase Chain Reaction Predictive Value of Tests *Quality
Assurance, Health Care MEETING ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).