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DNA Bacteriophage as Controls for Clinical Viral Testing

¹ Ambion, Inc., Austin, TX

^aAddress correspondence to this author at: Ambion, Inc., Diagnostics Division, 2170 Woodward St., Austin, TX 78744. Fax 512-651-0601; e-mail cwalkerpeach@ambion.com.

The first 300 words of the full text of this article appear below.

In the mid- to late-1980s, a revolution in molecular diagnostics began with the introduction of innovative methods for the detection of nucleic acids. In retrospect, the appearance of these technologies roughly coincided with the debuts of new pathogenic viruses, such as human immunodeficiency virus (HIV) and hepatitis C virus (HCV). Techniques such as PCR, transcription-mediated amplification (TMA), and branched DNA (bDNA) were applied to the detection and quantification of these viruses. Eventually, these tests were integrated into routine clinical laboratories for diagnosing and monitoring the treatment of patients infected with these viruses. Quantification data ("viral load") indicated whether the current drug cocktail was having the desired effect on the virus (i.e., lowering the viral load). If not, then another drug course could be prescribed. Today, the main advantages of these assays are great sensitivity (measuring as few as 50 copies/mL of plasma), ease of use for quantification, and early detection of viral nucleic acid in the peripheral blood before an antibody response develops, an application that has proven to be especially important in screening of human blood products.

These technologies were focused not only on the newly emerging RNA viruses but also on the better-known hepatitis B virus (HBV), cytomegalovirus (CMV), herpes simplex virus (HSV), varicella zoster virus (VZV), human papillomavirus (HPV), and Epstein–Barr virus (EBV). In developing DNA-based tests for these viruses, laboratories incorporated positive controls from one of three potential sources: plasmid DNA, positive patient specimens, or commercially available viral preparations. None of these formats is ideal. Plasmid DNA cannot be used until after the specimen analyte has been extracted. Patient specimens have become more difficult to use in the US after the introduction of new "HIPAA" regulations for protection of patient privacy. Moreover, viral nucleic acids in patient specimens are degraded during multiple freeze–thaw cycles. Lastly, the . . . [Full Text of this Article]

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