Comparison of Selective Polymerase Chain Reaction Primers and Differential Probe Hybridization of Polymerase Chain Reaction Products for Determination of Relative Amounts of Codon 215 Mutant and Wild-Type HIV-1 Populations

Eastman, Peter; Urdea, Mickey S.; Besemer, Diana J.; Stempien, Michelle M.; Kolberg, Janice A.

doi:10.1097/00042560-199507000-00007

Cited by 13 publications

(12 citation statements)

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“…Most likely, if we were to use highly sensitive detection techniques, such as ligase chain reaction [34], allele-specific PCR [35], or terminal dilution PCR [36,37], we would detect the persistence of NNRTI resistance in blood and semen for even longer [38]. Additionally, given that SP viral loads are generally lower than BP viral loads, as reviewed by Coombs et al [40], population-based sequencing may be more susceptible to sampling bias [41].…”

Section: Discussionmentioning

confidence: 99%

Long‐Term Persistence of Transmitted HIV Drug Resistance in Male Genital Tract Secretions: Implications for Secondary Transmission

et al. 2007

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Background. Transmitted drug-resistant HIV slowly reverts in the blood to drug-sensitive virus. The environment of the male genital tract (MGT) may result in even slower rates of reversion to drug susceptibility.Methods. We measured the decay of resistance in longitudinally collected blood and semen samples from 5 individuals newly infected with HIV containing resistance mutations to nonnucleoside reverse-transcriptase inhibitors (NNRTIs). We also investigated the sexual transmission of HIV to and from these participants.Results. In 3 of the 5 individuals, NNRTI resistance persisted in blood and semen throughout follow-up (mean, 296 days after the estimated day of infection [EDI]). In the other 2 individuals, NNRTI resistance persisted in blood and semen for 871 and 1179 days after the EDI; however, even after NNRTI resistance had fully reverted in blood, it remained readily detectable in semen. Two transmission groups were identified among these participants-one as the recipient partner and the other as the source partner.Conclusions. Transmitted drug-resistant HIV, which persists in blood for years, may revert to wild type even more slowly in the MGT. This prolonged persistence in the MGT may contribute to the high prevalence rates of transmitted drug resistance.

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Section: Discussionmentioning

confidence: 99%

Long‐Term Persistence of Transmitted HIV Drug Resistance in Male Genital Tract Secretions: Implications for Secondary Transmission

et al. 2007

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“…To date, several phenotypic and genotypic assays have been developed and are being used to monitor drug resistance (6). Genotypic assays are based on the detection of resistance-related mutations and provide indirect evidence of resistance (2,3,17,19). Phenotypic assays measure the ability of the virus to replicate in the presence of a drug and thus provide a direct measurement of drug susceptibility.…”

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confidence: 99%

Comparative Analysis of Two Commercial Phenotypic Assays for Drug Susceptibility Testing of Human Immunodeficiency Virus Type 1

et al. 2002

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Human immunodeficiency virus type 1 (HIV-1) isolates from 50 plasma specimens were analyzed for phenotypic susceptibility to licensed reverse transcriptase inhibitors and protease inhibitors by the Antivirogram and PhenoSense HIV assays. Twenty of these specimens were from recently seroconverted drug-naïve persons, and 30 were from patients who were the sources of occupational exposures to HIV-1; 16 of the specimens in the latter group were from drug-experienced patients. The phenotypic results of the Antivirogram and PhenoSense HIV assays were categorized as sensitive or reduced susceptibility on the basis of the cutoff values established by the manufacturers of each assay. Data for 12 to 15 drugs were available by both assays for 38 specimens and represented a total of 529 pairs of results. The two data sets had a 91.5% concordance by phenotypic category. The discordant results (n ‫؍‬ 45) were distributed randomly among 26 specimens and included 28 results (62.2%) which were within a twofold difference of the assay cutoff values. None of the discordant results were associated with primary resistance mutations that predicted high-level (>20-fold) resistance. Discordant results were distributed equally among specimens from drug-experienced and drugnaïve individuals and were slightly higher for protease inhibitors than for nonnucleoside reverse transcriptase inhibitors or nucleoside reverse transcriptase inhibitors. The findings of the present study demonstrate that the results of the Antivirogram and PhenoSense HIV assays correlate well, despite the use of different testing strategies.

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“…A PCRbased nonisotopic ligase detection reaction has been described (5), but it requires visual interpretation of the results. We have previously described a PCR-based differential probe hybridization strategy for the determination of the relative amounts of codon 215 mutant (MUT) and wild-type (WT) species in plasma virus RNA (4,6). This procedure employs 32 P-labeled probes and an AMBIS 4000 Image Analyzer for quantification of hybridization.…”

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confidence: 99%

“…In order to determine the sensitivity of the nonisotopic detection methods for detection of small amounts of MUT, mixtures of codon 215 MUT and WT HIV RNA were PCR amplified and analyzed isotopically as previously described (4) and nonisotopically. The results presented in Fig.…”

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confidence: 99%

“…Colorimetric analysis of a dilution series of an in vitro transcript of cloned HIV strain SF2 (14) which is WT at codon 215 resulted in a mean MUT/WT ratio of 0.018 (standard deviation, 0.011), while chemiluminescent detection produced an MUT/WT ratio of 0.009 (standard deviation, 0.002). A similar dilution series of a MUT sample (patient 6, day 159 [4]) produced a colorimetric mean MUT/WT ratio of 28.7 (standard deviation, 6.0), while chemiluminescent detection of a MUT control from the National Institutes of Health (Bethesda, Md.) AIDS Research and Reference Program (catalog 1074) produced a mean MUT/WT ratio of 99.1 (standard deviation, 7.8).…”

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Nonisotopic hybridization assay for determination of relative amounts of genotypic human immunodeficiency virus type 1 zidovudine resistance

Eastman¹,

Boyer²,

Molé³

et al. 1995

J Clin Microbiol

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A nonisotopic hybridization assay for human immunodeficiency virus genotypic zidovudine resistance determination is described. Biotinylated PCR product was hybridized with enzyme-labeled probes for wild-type or resistant mutant sequences and detected colorimetrically or chemiluminescently in a microplate format. Changes in mutant-to-wild-type ratios allow for the monitoring of longitudinal patient samples.

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Comparison of Selective Polymerase Chain Reaction Primers and Differential Probe Hybridization of Polymerase Chain Reaction Products for Determination of Relative Amounts of Codon 215 Mutant and Wild-Type HIV-1 Populations

Cited by 13 publications

References 0 publications

Long‐Term Persistence of Transmitted HIV Drug Resistance in Male Genital Tract Secretions: Implications for Secondary Transmission

Long‐Term Persistence of Transmitted HIV Drug Resistance in Male Genital Tract Secretions: Implications for Secondary Transmission

Comparative Analysis of Two Commercial Phenotypic Assays for Drug Susceptibility Testing of Human Immunodeficiency Virus Type 1

Nonisotopic hybridization assay for determination of relative amounts of genotypic human immunodeficiency virus type 1 zidovudine resistance

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