TETALI S, WANG XP, ROMANO JW, LEE EM, KAPLAN MH, HAIMA P, GINOCCHIO CC; Interscience Conference on Antimicrobial Agents and Chemotherapy.

Abstr Intersci Conf Antimicrob Agents Chemother Intersci Conf Antimicrob Agents Chemother. 1999 Sep 26-29; 39: 232 (abstract no. 1589).

North Shore Univ. Hosp., Manhasset, NY

BACKGROUND: Several studies have demonstrated that resistance to HIV-1 infection and delayed disease progression are often linked to a 32 base pair deletion (Delta 32) in the CCR5 chemokine receptor, which serves as a cellular entry cofactor for macrophage tropic HIV-1 isolates.METHODS: A NASBA based CCR5 genotyping assay was modified for testing with the NucliSens Basic Kit (Organon Teknika, NL). CCR5 allele status (wild type, homozygous Delta 32, heterozygous Delta 32) was determined using PBMCs, whole blood, dried blood spots and/or buccal scrapings from 76 patients. NASBA results were confirmed using a PBMC based DNA PCR assay.RESULTS: The CCR5 genotype patterns obtained by NASBA from 50 ul dried whole blood spots and buccal scrapings were identical with NASBA PBMC and whole blood specimens. Results from all 4 specimen types were 100% concordant with the PBMC DNA PCR results. In all, 59 patients (78 %) were homozygous wild type, 16 patients (21 %) were heterozygous Delta 32 and 1 patient (1 %) demonstrated a homozygous Delta 32 CCR5 genotype. The Delta 32 homozygous person remains HIV-1 seronegative after many years of exposure to HIV through IV drug use and heterosexual contact with her HIV-1 infected spouse. We also identified 3 CCR5 homozygous wild type HIV-1 seronegative persons with multiple exposures to HIV-1 through IV drug use. This data suggests the presence of mechanisms of resistance to HIV-1 infection independent of CCR5 allele status.CONCLUSIONS: This study demonstrates that NASBA is a simple, highly sensitive, and specific assay for CCR5 genotyping using a variety of specimen types. The ease of collection of buccal scrapings and dried blood spots permits the use of this assay for large epidemiology studies without the expense and time required for assays that use PBMCs.

Publication Types:

• Meeting Abstracts

Keywords:

• Acquired Immunodeficiency Syndrome
• Alleles
• Chemokines
• Disease Progression
• Exanthema
• Genotype
• HIV Infections
• HIV Seropositivity
• HIV-1
• Humans
• Polymerase Chain Reaction
• Proto-Oncogene Proteins c-kit
• Receptors, CCR5
• blood
• genetics
• immunology
• transmission

Other ID:

• GWAIDS0007729

UI: 102245226

From Meeting Abstracts