Journal Article

Isolation of SARS-CoV-2 in Viral Cell Culture in Immunocompromised Patients With Persistently Positive RT-PCR Results

Reske, Kimberly A., Fraser, Victoria J., Babcock, Hilary M., O’Neil, Caroline A., Diamond, Michael S., Kwon, Jennie H., McDonald, David, Bailey, Adam L., Burnham, Carey-Ann D., Peacock, Kate, Sung, Abby, Wallace, Meghan A., Stewart, Henry B., Miller, Candace

2022

Overview

Immunocompromised adults can have prolonged acute respiratory syndrome coronavirus 2 (SARS-CoV-2) positive RT-PCR results, long after the initial diagnosis of coronavirus disease 2019 (COVID-19). This study aimed to determine if SARS-CoV-2 virus can be recovered in viral cell culture from immunocompromised adults with persistently positive SARS-CoV-2 RT-PCR tests. We obtained 20 remnant SARS-CoV-2 PCR positive nasopharyngeal swabs from 20 immunocompromised adults with a positive RT-PCR test ≥14 days after the initial positive test. The patients’ 2 nd test samples underwent SARS-CoV-2 antigen testing, and culture with Vero-hACE2-TMPRSS2 cells. Viral RNA and cultivable virus were recovered from the cultured cells after qRT-PCR and plaque assays. Of 20 patients, 10 (50%) had a solid organ transplant and 5 (25%) had a hematologic malignancy. For most patients, RT-PCR Ct values increased over time. There were 2 patients with positive viral cell cultures; one patient had chronic lymphocytic leukemia treated with venetoclax and obinutuzumab who had a low viral titer of 27 PFU/mL. The second patient had marginal zone lymphoma treated with bendamustine and rituximab who had a high viral titer of 2 x 10 6 PFU/mL. Most samples collected ≥7 days after an initial positive SARS-CoV-2 RT-PCR had negative viral cell cultures. The 2 patients with positive viral cell cultures had hematologic malignancies treated with chemotherapy and B cell depleting therapy. One patient had a high concentration titer of cultivable virus. Further data are needed to determine risk factors for persistent viral shedding and methods to prevent SARS-CoV-2 transmission from immunocompromised hosts.

• Publisher: Frontiers Media SA,Frontiers Media S.A
• Subject: Antigens / Bone marrow / Buffalo / Cell culture / Cell Culture Techniques / Cellular and Infection Microbiology / Chemotherapy / Chronic lymphocytic leukemia / Coronaviruses / COVID-19 / Demographics / Disease transmission / Emergency medical care / Humans / immunocompromised / Immunocompromised Host / Immunocompromised hosts / laboratory medicine / Lymphatic leukemia / Lymphoma / Malignancy / Medical laboratories / obinutuzumab / Patients / Polymerase chain reaction / Reverse Transcriptase Polymerase Chain Reaction / Risk factors / Rituximab / RNA viruses / RNA, Viral - genetics / SARS-CoV-2 / Severe acute respiratory syndrome coronavirus 2 / Transplants & implants