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. 2021 Aug:294:114182.
doi: 10.1016/j.jviromet.2021.114182. Epub 2021 May 10.

Evaluation of the analytical performance and specificity of a SARS-CoV-2 transcription-mediated amplification assay

Markus Schneider  1 , Thomas Iftner  1 , Tina Ganzenmueller  2
Affiliations

Evaluation of the analytical performance and specificity of a SARS-CoV-2 transcription-mediated amplification assay

Markus Schneider et al. J Virol Methods. 2021 Aug.

Abstract

The ongoing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic requires fast and accurate high-throughput diagnostic tools. To evaluate the analytical performance of the Hologic Aptima transcription-mediated amplification (TMA) assay for detection of SARS-CoV-2 RNA from respiratory samples we analysed 103 clinical and proficiency panel samples pre-tested by real-time RT-PCR (Altona, RealStar) and found a positive percent agreement (sensitivity) of 95.7 % and a negative percent agreement (specificity) of 100 %. The limit of detection of the Aptima test was 150 copies/mL determined as 95 % detection probability. To further assess the Aptima assay's specificity we prospectively analysed 7545 clinical specimens from the upper and lower respiratory tract sent for the purpose of routine SARS-CoV-2 screening. SARS-CoV-2 RNA was detected in 16/7545 (0.2 %) samples by the TMA assay and confirmed independently by the Xpert SARS-CoV-2 RT-PCR (Cepheid); in one case a previous discrepant result was confirmed as true SARS-CoV-2 infection in a subsequent sample from the same patient. Results from the Aptima SARS-CoV-2 TMA assay agreed well with RT-PCR and showed an excellent specificity in a large number of routine specimens despite the low prevalence at that time of the pandemic, indicating that this assay can be used even for screening purposes.

Keywords: Aptima; Limit of detection; SARS-CoV-2; Screening; Specificity; Transcription-mediated amplification.

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Conflict of interest statement

The University Hospital Tuebingen (Thomas Iftner) received an unconditional research grant from Hologic for a longitudinal study on a comparison of HPV tests for cervical cancer screening in the past. The other authors report no conflict of interest. Hologic sponsored the first 250 tests of the evaluation study; otherwise the study was financed by institutional funds of the Virology Department of the University Hospital Tuebingen. Hologic did not have any influence on the scientific data interpretation or content of the submitted manuscript.

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