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?:abstract
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OBJECTIVES The purpose of this study was to detect COVID-19 cases with persistent positive RT-PCR results for SARS-CoV-2, for which viable virus can be inferred, due to the presence of subgenomic (SG) viral RNA, which is expressed only in replicating viruses. METHODS RNA remnants, purified from diagnostic nasopharyngeal specimens, were used as templates for RT-PCR specific detection of SG E gene RNA. As controls, we also detected viral genomic RNA for the E gene and/or a human housekeeping gene (RNase P). RESULTS We assessed the samples of 60 RT-PCR-positive cases with a prolonged viral SARS-CoV-2 shedding (24-101 days) since the first diagnostic RT-PCR. SG viral RNA was detected in 12/60 (20%) of the persistent cases, 28-79 days after the onset of symptoms. The age range of the cases with prolonged viral shedding and presence of SG RNA was quite wide (40-100 years), and they were equally distributed between males (42%) and females (58%). None was HIV positive, although seven were immunosuppressed. According to the severity of the COVID-19 episodes they were mild (40%), intermediate (20%), and severe (40%)Conclusion: In a percentage of persistent positive SARS-CoV-2 PCR positive cases the presence of actively replicating virus may be inferred, far beyond diagnosis. We should not assume a universal lack of infectiousness for COVID-19 cases with prolonged viral shedding.
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