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).