LOW COST HIV DRUG RESISTANCE MONITORING

描述：(摘自申请者摘要)Jack U‘Ren博士，Paul Haydock和Sharon Howard(Saigene Corporation[正式诊断 MicroProbe Corporation]，Bothell，WA)建议开发 一种低成本的艾滋病毒耐药监测方法。这款产品既可以 快速量化病毒载量并识别艾滋病毒反向突变 转录酶基因。目前，还没有一种产品可以同时衡量这两种情况 病毒载量和鉴定耐药突变的同时 同样的样本。总体目标将通过执行 定量聚合酶链式反应检测已知突变体的探针结合 HIV RT基因中的位置。在第一阶段，化验将发展为 通过重点研究聚合酶链式反应条件、引物、捕获序列和 定量检测野生型和毒品的内部标准- 具有抗药性的艾滋病毒。然后，将使用以下方法对开发的化验进行评估 具有抗药性的艾滋病毒毒株的特征。在第二阶段，化验将是 扩展到其他药物和药物组合，并将在临床上使用 样本。这些结果将与成本更高的DNA测序进行比较 方法在正在进行的临床研究中。终极临床应用 将为患者管理提供耐药性信息， 因此，了解耐药突变的确切性质可能会 预测对其他药物的交叉耐药性或耐药性降低。

DESCRIPTION: (Adapted from applicant's abstract) Drs. Jack U'Ren, Paul Haydock and Sharon Howard (Saigene Corporation [formally the Diagnostic Division of the MicroProbe Corporation], Bothell, WA) propose to develop a low cost HIV drug-resistant monitoring assay. This product will both rapidly quantify viral load and identify mutations in the HIV reverse transcriptase gene. At present, no products exist which measure both viral load and identify drug-resistant mutations at the same time from the same sample. The overall aim will be accomplished by performing quantitative PCR followed by examining probe binding at known mutant locations within the HIV RT gene. In phase I, the assay will develop be developed by focusing on PCR conditions, primers, capture sequences, and internal standards which quantitatively detect wild type and drug- resistant HIV. The developed assay will then be evaluated using characterized drug-resistant HIV strains. In Phase II, the assay will be expanded to other drugs and drug combinations, and will employ clinical samples. These results will be compared to more costly DNA sequencing methods in ongoing clinical studies. The ultimate clinical application will be to provide drug resistance information for patient management, such that knowing the precise nature of a drug resistant mutation may predict cross-resistance or reduced resistance to other drugs.