Repeated Coronavirus Disease 2019 Molecular Testing: Correlation of Severe Acute Respiratory Syndrome Coronavirus 2 Culture With Molecular Assays and Cycle Thresholds.
Repeated Coronavirus Disease 2019 Molecular Testing: Correlation of Severe Acute Respiratory Syndrome Coronavirus 2 Culture With Molecular Assays and Cycle Thresholds.
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DOI：
10.1093/cid/ciaa1616
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发表时间：
2021-08-16
期刊：
影响因子：
--
通讯作者：
Mostafa HH
中科院分区：
文献类型：
--
作者：
Gniazdowski V;Paul Morris C;Wohl S;Mehoke T;Ramakrishnan S;Thielen P;Powell H;Smith B;Armstrong DT;Herrera M;Reifsnyder C;Sevdali M;Carroll KC;Pekosz A;Mostafa HH
重复的2019冠状病毒病（COVID-19）分子检测可能导致阴性结果后的阳性结果，并随着时间的推移导致多次阳性结果。检测阳性结果与传染性病毒之间的关系很重要。采用为期2个月的回顾性队列数据和连续采集的COVID-19患者或调查患者标本，了解病毒RNA延长阳性检测结果、周期阈值（Ct）值与细胞培养中严重急性呼吸综合征冠状病毒2 （SARS-CoV-2）生长的相关性。在病毒RNA检测时间延长的患者中，采用全基因组测序来确认病毒基因型。采用液滴数字聚合酶链反应评估新冠肺炎诊断检测结果假阴性率。2个月内共检测标本29686份，重复检测2194例。在细胞培养中，SARS-CoV-2靶基因的平均Ct值为18.8（3.4）。在首次阳性结果后21天内收集的标本中，延长的病毒RNA脱落与培养中的病毒阳性生长有关，但主要发生在样本收集时有症状的个体中。全基因组测序提供了同样的病毒随时间携带的证据。阴性结果后的阳性试验结果的Ct值为>29.5，与病毒培养无关。从新冠肺炎确诊或临床疑似病例采集的阴性标本中，液滴数字聚合酶链反应结果为阳性的占5.6%。SARS-CoV-2诊断试验的低Ct值与细胞培养中的病毒生长有关。有症状且病毒RNA脱落时间延长的患者也可能具有传染性。在病毒RNA脱落时间延长和阴性结果后呈阳性的情况下，评估了严重急性呼吸综合征冠状病毒2诊断周期阈值与细胞培养中病毒生长之间的相关性。液滴数字聚合酶链反应用于调查假阴性率。
Repeated coronavirus disease 2019 (COVID-19) molecular testing can lead to positive test results after negative results and to multiple positive results over time. The association between positive test results and infectious virus is important to quantify. A 2-month cohort of retrospective data and consecutively collected specimens from patients with COVID-19 or patients under investigation were used to understand the correlation between prolonged viral RNA positive test results, cycle threshold (Ct) values and growth of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in cell culture. Whole-genome sequencing was used to confirm virus genotype in patients with prolonged viral RNA detection. Droplet digital polymerase chain reaction was used to assess the rate of false-negative COVID-19 diagnostic test results. In 2 months, 29 686 specimens were tested and 2194 patients underwent repeated testing. Virus recovery in cell culture was noted in specimens with a mean Ct value of 18.8 (3.4) for SARS-CoV-2 target genes. Prolonged viral RNA shedding was associated with positive virus growth in culture in specimens collected up to 21 days after the first positive result but mostly in individuals symptomatic at the time of sample collection. Whole-genome sequencing provided evidence the same virus was carried over time. Positive test results following negative results had Ct values >29.5 and were not associated with virus culture. Droplet digital polymerase chain reaction results were positive in 5.6% of negative specimens collected from patients with confirmed or clinically suspected COVID-19. Low Ct values in SARS-CoV-2 diagnostic tests were associated with virus growth in cell culture. Symptomatic patients with prolonged viral RNA shedding can also be infectious. A correlation between severe acute respiratory syndrome coronavirus 2 diagnostic cycle threshold values and virus growth in cell culture is assessed in the context of prolonged viral RNA shedding and positive after negative results. Droplet digital polymerase chain reaction is used to investigate rates of false-negativity.
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