Clinical performance of non-radioactive assays for HIV-1 DNA amplified by the polymerase chain reaction

Abstract

A major factor preventing more widespread use of polymerase chain reaction in the clinical laboratory is the lack of convenient non-radioactive probe hybridization procedures which do not sacrifice sensitivity or specificity. In this report, we describe comparisons of probes labelled with biotin, digoxygenin, alkaline phosphatase, and(32)P. We report the comparison of solution or liquid hybridization assay and Southern blotting with digoxygenin-labelled oligonucleotides on a total of 64 clinical specimens. Perfect diagnostic agreement between the(32)P and digoxygenin probes was obtained. These data suggest that the non-radioactive assay as described is as sensitive and as specific as the assay with(32)P-Iabelled probes.

A flexible bioluminescent‐quantitative polymerase chain reaction assay for analysis of competitive PCR amplicons

“…The chemiluminescent-based labeling systems have held great promise for assay development (19). These systems include oligonucleotide probes labeled with acridinium esters (20), horseradish peroxidase (21), alkaline phosphatase (22), and digoxigenin (23)(24)(25)(26). While the acridinium ester-based homogenous assays are not providing the sensitivity required for all applications, the enzyme-based systems have been used to generate assays with sensitivities greater than what can be attained with radioisotopes.…”