a sensitive branched dna hiv-1 signal amplification viral load assay with single day turnaround一个敏感支dna hiv - 1信号放大病毒载量测定一天好转.pdfVIP

a sensitive branched dna hiv-1 signal amplification viral load assay with single day turnaround一个敏感支dna hiv - 1信号放大病毒载量测定一天好转.pdf

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a sensitive branched dna hiv-1 signal amplification viral load assay with single day turnaround一个敏感支dna hiv - 1信号放大病毒载量测定一天好转

A Sensitive Branched DNA HIV-1 Signal Amplification Viral Load Assay with Single Day Turnaround Mark A. Baumeister, Nan Zhang, Hilda Beas, Jesse R. Brooks, Jesse A. Canchola, Carlo Cosenza, Felix Kleshik, Vinod Rampersad, Johan Surtihadi, Thomas R. Battersby* Siemens Healthcare Diagnostics Inc., Berkeley, California, United States of America Abstract Branched DNA (bDNA) is a signal amplification technology used in clinical and research laboratories to quantitatively detect nucleic acids. An overnight incubation is a significant drawback of highly sensitive bDNA assays. The VERSANTH HIV-1 RNA 3.0 Assay (bDNA) (‘‘Versant Assay’’) currently used in clinical laboratories was modified to allow shorter target incubation, enabling the viral load assay to be run in a single day. To dramatically reduce the target incubation from 16–18 h to 2.5 h, composition of only the ‘‘Lysis Diluent’’ solution was modified. Nucleic acid probes in the assay were unchanged. Performance of the modified assay (assay in development; not commercially available) was evaluated and compared to the Versant Assay. Dilution series replicates (.950 results) were used to demonstrate that analytical sensitivity, linearity, accuracy, and precision for the shorter modified assay are comparable to the Versant Assay. HIV RNA-positive clinical specimens (n = 135) showed no significant difference in quantification between the modified assay and the Versant Assay. Equivalent relative quantification of samples of eight genotypes was demonstrated for the two assays. Elevated levels of several potentially interfering endogenous substances had no effect on quantification or specificity of the modified assay. The modified assay with drastically improved turnaround time demonstrates the viability of signal-amplifying technology, such as bDNA, as an alternat

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