NASBA: A Novel, Isothermal Detection Technology for Qualitative and Quantitative HIV-1 RNA Measurements

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Although immunoassays have long served as the standard in the field of diagnostics, the advent of nucleic acid amplification technologies allows for a new array of diagnostic applications. NASBA, nucleic acid sequence-based amplification, is one such technology that is highly suited for the amplification of RNA. As such, NASBA is applied readily as a diagnostic tool for infectious diseases, particularly for RNA viruses, such as retroviruses. The development and application of NASBA technology as a qualitative and quantitative diagnostic system for HIV-1 are described in this article.

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    We have reported a similar method for calculating cut-off values [9,10,15]. The use of appropriate internal controls for quantifying HIV viral load using the NASBA technique has been described [14] and the technique is applicable to other targets. The dynamic range of the ECL detector was determined using a serial dilution of a known positive sample of avian influenza H5N1.

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    Other users of NASBA systems have reported similar methods for calculating the cut-off value, including 0.01–0.025×IRS [25,36,37] and 200 ECL units [38]. The use of appropriate internal controls for quantifying HIV viral load using the NASBA technique has been described [14] and the technique is applicable to other targets. Analysing the same sample multiple times assessed reproducibility.

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