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M94A3316.TXT
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1994-10-25
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Document 3316
DOCN M94A3316
TI Detection of reverse transcriptase (RT) codon 215 mutations by PCR in
HIV-infected adults: correlation with clinical disease.
DT 9412
AU Nelson R; Bradley W; Day N; Good R; Univ. South Florida/All Children's
Hospital, St. Petersburg; 33701.
SO Int Conf AIDS. 1994 Aug 7-12;10(1):104 (abstract no. PA0033). Unique
Identifier : AIDSLINE ICA10/94369259
AB OBJECTIVE: To rapidly detect mutations of the HIV-1 reverse
transcriptase codon 215 (associated with azidothymidine resistance) from
peripheral blood DNA of disease-progressing and non-progressing adult
patients. METHODS: Thirty-two peripheral blood mononuclear cell samples
were obtained from 29 HIV-infected adults. Cell DNA was isolated and RT
gene and codon 215-specific oligonucleotides were used in an
amplification-refractory-to-mutation assay (ARMS) to detect the presence
of wild type threonine (Thr) or the common mutation,
tyrosine/phenylalanine (Tyr/Phe). Patients were categorized as
AZT-resistant (duration of AZT > 18 mo, drop in CD4# > 50%, increase in
beta-2 microglobulin (B2M) > 1mg/dl, weight loss > 10% body mass) or
AZT-sensitive (duration of AZT < 4 mo, stable CD4#, B2M, weight).
Differences between the prevalence of codon 215 mutations were compared.
RESULTS: Fifteen AZT-sensitive samples were analyzed utilizing the
primers for human beta-globin, HIV V3, codon 215-Thr, and codon
215-Tyr/Phe. Eight of 15 (53.3%) samples had > 50% of the PCR products
derived from 215-Thr, 2 samples had > 50% of the product derived from
215-Tyr/Phe, 5 samples failed to generate a V3- or codon 215-product.
Fourteen of 17 (82.4%) AZT-resistant samples had > 50% of the PCR
products derived from 215-Tyr/Phe. Two samples failed to amplify HIV V3
and one which did amplify HIV V3 gave equivicol codon 215 results.
DISCUSSION AND CONCLUSIONS: ARMS PCR is a useful test to rapidly detect
codon 215 mutations (Thr - or > Tyr/Phe) associated with AZT resistance.
Eighty-two percent of patients with evidence for clinical resistance and
20% without evidence of clinical resistance had detectable 215-Tyr/Phe
mutations. Twenty-five percent of the samples failed to amplify with
either primer (215-Thr or 215-Tyr/Phe).
DE Adult Codon/GENETICS Drug Resistance, Microbial/GENETICS DNA
Primers/GENETICS DNA, Viral/BLOOD/GENETICS Human HIV Envelope Protein
gp120/GENETICS HIV Infections/DRUG THERAPY/*MICROBIOLOGY HIV-1/DRUG
EFFECTS/*ENZYMOLOGY/*GENETICS Peptide Fragments/GENETICS *Point
Mutation Polymerase Chain Reaction/METHODS Prognosis Reverse
Transcriptase/*GENETICS Time Factors Zidovudine/THERAPEUTIC USE
MEETING ABSTRACT
SOURCE: National Library of Medicine. NOTICE: This material may be
protected by Copyright Law (Title 17, U.S.Code).