Comparison of capillary electrophoresis sequencing with the new CEQ 2000 DNA Analysis System to conventional gel based systems for HIV drug resistance analysis

被引：24

作者：

Merel, P ^{[1
]}

Pellegrin, I ^{[1
]}

Garrigue, I ^{[1
]}

Caumont, A ^{[1
]}

Schrive, MH ^{[1
]}

Birac, V ^{[1
]}

Bonot, P ^{[1
]}

Fleury, H ^{[1
]}

机构：

[1] Ctr Hosp Reg & Univ Victor Segalen, Virol Lab, Bordeaux, France

来源：

JOURNAL OF VIROLOGICAL METHODS | 2001年 / 98卷 / 01期

关键词：

HIV-1; genotyping; capillary electrophoresis; sequencing; cost-effectiveness;

D O I：

10.1016/S0166-0934(01)00338-X

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

To date the majority of sequencing technologies have been based on use of gel plates. In this study sequencing by capillary electrophoresis for HIV-1 genotyping on the CEQ 2000 sequencer (Beckman Coulter Inc.) has been investigated and compared to an 'in house' protocol on the Prism-377 sequencer (Applied Biosystems) and to the HIV-1 TruGene kit (Visible Genetics Inc.), two gel plate-based systems. Plasma from 20 HAART-treated patients with virological failure were analyzed for protease (PR) and reverse transcriptase (RT) genes. A total of 520 RT codons (26/patient) and 360 PR codons (18/patient) related to antiretroviral drug resistance were evaluated. The overall agreement between CEQ 2000 and Prism-377 results was 100% for the RT and PR primary and secondary mutations. The overall agreement between CEQ 2000 and TruGene was 100% for primary and greater than or equal to 97% for secondary mutations. Discrepant results would have never led to errors in genotype interpretation. Performances for a 24 patients/week/one technician genotyping throughput were analyzed. For Prism-377, TruGene and CEQ 2000, manual processing required 5, 4 and 2,5 days, sequence data analysis needed additional 3, 1 and 2 days and cost/patient was approximate to 49, 214 and 39 $, respectively. The CEQ 2000 sequencer offers a reliable alternative for fast and cost effective HIV drug resistance analysis. (C) 2001 Elsevier Science B.V. All rights reserved.

引用

页码：9 / 16

页数：8

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