NucliSens EasyQ HIV-1 V1.2 system: Detection of human plasma-derived background signal

Until recently the NucliSens EasyQ HIV-1 V1.2 system has been used throughout South Africa as part of the national antiretroviral roll-out programme for the monitoring of HIV-1 RNA load in patients on antiretroviral treatment. Shortly after changing to a new assay lot number an increased proportion of patient specimens, showing detectable but low viral loads, was observed (<200 IU/ml). The test runs remained valid as the lysis buffer-only no-template controls (NTCs) remained negative. Contamination with amplification product was excluded. Subsequently the same phenomenon was observed in at least three other South African laboratories across different assay lot numbers. When testing aliquots of plasma, freshly obtained from HIV-negative donors, at two of these laboratories, 33/134 aliquots showed detectable values (range 26-370, median: 64 IU/ml), while all NTCs remained negative. These findings emphasize the importance of appropriate specimen controls in all diagnostic assays. In this case HIV-negative human plasma should be included routinely in addition to NTCs, which would allow rapid detection of a background signal.