Open Access

Sensitivity assessment of droplet digital PCR for SARS-CoV-2 detection

  • Authors:
    • Luca Falzone
    • Nicolò Musso
    • Giuseppe Gattuso
    • Dafne Bongiorno
    • Concetta Ilenia Palermo
    • Guido Scalia
    • Massimo Libra
    • Stefania Stefani
  • View Affiliations

  • Published online on: July 13, 2020     https://doi.org/10.3892/ijmm.2020.4673
  • Pages: 957-964
  • Copyright: © Falzone et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) is the gold standard method for the diagnosis of COVID‑19 infection. Due to pre‑analytical and technical limitations, samples with low viral load are often misdiagnosed as false‑negative samples. Therefore, it is important to evaluate other strategies able to overcome the limits of RT‑qPCR. Blinded swab samples from two individuals diagnosed positive and negative for COVID‑19 were analyzed by droplet digital PCR (ddPCR) and RT‑qPCR in order to assess the sensitivity of both methods. Intercalation chemistries and a World Health Organization (WHO)/Center for Disease Control and Prevention (CDC)‑approved probe for the SARS‑CoV‑2 N gene were used. SYBR‑Green RT‑qPCR is not able to diagnose as positive samples with low viral load, while, TaqMan Probe RT‑qPCR gave positive signals at very late Ct values. On the contrary, ddPCR showed higher sensitivity rate compared to RT‑qPCR and both EvaGreen and probe ddPCR were able to recognize the sample with low viral load as positive even at 10‑fold diluted concentration. In conclusion, ddPCR shows higher sensitivity and specificity compared to RT‑qPCR for the diagnosis of COVID‑19 infection in false‑negative samples with low viral load. Therefore, ddPCR is strongly recommended in clinical practice for the diagnosis of COVID‑19 and the follow‑up of positive patients until complete remission.

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September-2020
Volume 46 Issue 3

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Spandidos Publications style
Falzone L, Musso N, Gattuso G, Bongiorno D, Palermo CI, Scalia G, Libra M and Stefani S: Sensitivity assessment of droplet digital PCR for SARS-CoV-2 detection. Int J Mol Med 46: 957-964, 2020
APA
Falzone, L., Musso, N., Gattuso, G., Bongiorno, D., Palermo, C.I., Scalia, G. ... Stefani, S. (2020). Sensitivity assessment of droplet digital PCR for SARS-CoV-2 detection. International Journal of Molecular Medicine, 46, 957-964. https://doi.org/10.3892/ijmm.2020.4673
MLA
Falzone, L., Musso, N., Gattuso, G., Bongiorno, D., Palermo, C. I., Scalia, G., Libra, M., Stefani, S."Sensitivity assessment of droplet digital PCR for SARS-CoV-2 detection". International Journal of Molecular Medicine 46.3 (2020): 957-964.
Chicago
Falzone, L., Musso, N., Gattuso, G., Bongiorno, D., Palermo, C. I., Scalia, G., Libra, M., Stefani, S."Sensitivity assessment of droplet digital PCR for SARS-CoV-2 detection". International Journal of Molecular Medicine 46, no. 3 (2020): 957-964. https://doi.org/10.3892/ijmm.2020.4673