Comparison of QIAamp HCV kit spin columns, silica beads, and phenol-chloroform for recovering human immunodeficiency virus type 1 RNA from plasma

Shafer, Robert W.; Levee, D J; Winters, Mark A.; Richmond, K L; Huang, De-Bin; Merigan, Thomas C.

doi:10.1128/jcm.35.2.520-522.1997

Cited by 41 publications

(8 citation statements)

References 8 publications

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“…It is possible that there is a bias in template-to-product ratio caused by random variation in the early stages of PCR, resulting in over-representation of a particular species. It has also been estimated that the efficiency of RNA extraction and cDNA synthesis is 1-10% [9]. Thus if the complete extract from 1 mL of blood with a viral load of 100 copies/mL is used then only between one and 10 molecules are being examined, and in practice the proportion of sample used is usually much smaller.…”

Section: Discussionmentioning

confidence: 99%

Genotypic antiretroviral drug resistance testing at low viral loads in the UK

Cane¹,

Kaye²,

Smit³

et al. 2008

HIV Medicine

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Antiretroviral drug resistance testing is recommended in HIV-1 infected patients failing therapy in order to inform treatment selection. Although guidelines and test manufacturers recommend a viral load of at least 500-1000 HIV-1 RNA copies/mL for genotypic resistance testing to be performed, prompt management of virological failure could benefit from testing at lower viral load levels. MethodsLaboratories undertaking genotypic resistance testing were asked to provide figures for the number of resistance tests undertaken at viral loads o2000 copies/mL, the success rates of such tests and the extent of resistance detected, all stratified for viral load levels. ResultsOf the replies received, most laboratories were attempting resistance testing at viral loads below the recommended guidelines, with variable success and outcomes. ConclusionsThis audit of current practice in the UK for undertaking genotypic resistance tests at viral loads o1000 copies/mL highlights the widespread use of such testing outside the British HIV Association guidelines.

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Section: Discussionmentioning

confidence: 99%

Genotypic antiretroviral drug resistance testing at low viral loads in the UK

Cane¹,

Kaye²,

Smit³

et al. 2008

HIV Medicine

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“…Genotypic analyses. The preparation of HIV RNA from patient plasma, the generation of the 1.1-kb PCR fragments carrying HIV RT, and analyses of the DNA sequences of these PCR fragments have been described previously (15,44). Briefly, genotypic analyses were performed with RT-PCR fragments carrying HIV RT genes generated from the patients' plasma samples taken at the maintenance-phase baseline, the 6-month time point, and when available, the 12month time point.…”

Section: Methodsmentioning

confidence: 99%

Genotypic and Phenotypic Characterization of Human Immunodeficiency Virus Type 1 Variants Isolated from AIDS Patients after Prolonged Adefovir Dipivoxil Therapy

Mulato

Lamy

Miller

et al. 1998

Antimicrob Agents Chemother

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Adefovir dipivoxil [bis(pivaloyloxymethyl)-ester prodrug], an orally bioavailable prodrug of adefovir [9-(2-phosphonylmethoxyethyl)adenine], is currently in phase III clinical trials for the treatment of human immunodeficiency virus (HIV). In vitro experiments demonstrated that either a K65R or a K70E mutation in HIV reverse transcriptase (RT) was selected in the presence of adefovir, conferring a 16- or 9-fold decrease in susceptibility to adefovir, respectively. Previous data demonstrated that patients receiving adefovir dipivoxil monotherapy (125 mg daily) for 12 weeks experienced a median decrease in HIV RNA levels of 0.5 log10 copies/ml and that resistance to adefovir dipivoxil did not arise during that period. In the present investigation, a further study was undertaken to investigate whether RT mutations developed among viruses from patients who completed the 12-week study and who opted to enroll in a maintenance phase of prolonged (6- to 12-month) adefovir dipivoxil therapy (120 mg daily). Concomitant treatment with antiretroviral agents was permitted during the maintenance phase. The median decreases in HIV RNA levels for patients who completed 6 or 12 months of maintenance-phase dosing were 0.6 and 1.14 log10 copies/ml, respectively. The reductions in the HIV RNA levels were similar among patients who received adefovir dipivoxil with or without concomitant treatment with antiretroviral agents. Viruses from 8 of 29 patients dosed for up to 12 months developed RT mutations that were not present at baseline; these mutations may have been related to adefovir dipivoxil therapy. Viruses from two of the eight patients developed the K70E mutation while the patients were on therapy, but none of the viruses from patients developed the K65R RT substitution. Despite the development of RT mutations, sustained reductions (6 to 12 months) in viral load (≥0.7 log10 copies/ml decrease from baseline) were observed in all eight patients.

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“…Our data show that the PVDF filter method is suitable for single‐unit NAT, in that it can be readily adapted to total robotic automation. The PVDF method was found to have a sensitivity equal to or in some cases slightly less than that of one of the most sensitive extraction methods, 19,20 Qiagen columns. In all cases, the sensitivities were within two serial dilutions of each other.…”

Section: Discussionmentioning

confidence: 90%