高度敏感的血浆严重急性呼吸综合征冠状病毒 2 核酸定量分析揭示了其潜在的临床价值。

Highly Sensitive Quantification of Plasma Severe Acute Respiratory Syndrome Coronavirus 2 RNA Sheds Light on its Potential Clinical Value.

机构信息

Assistance Publique Hôpitaux de Paris-Centre (AP-HP.Centre), Service de Microbiologie (Unité de virologie), Hôpital Européen Georges Pompidou, Paris, France.

Université de Paris, Centre de Recherche des Cordeliers, Sorbonne Université, Inserm, Functional Genomics of Solid Tumors laboratory, équipe labellisée Ligue Nationale contre le Cancer, Labex OncoImmunology, Paris, France.

出版信息

Clin Infect Dis. 2021 Nov 2;73(9):e2890-e2897. doi: 10.1093/cid/ciaa1196.

DOI:10.1093/cid/ciaa1196

PMID:32803231

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7454373/

Abstract

BACKGROUND

Coronavirus disease 2019 (COVID-19) is a global public health problem that has already caused more than 662 000 deaths worldwide. Although the clinical manifestations of COVID-19 are dominated by respiratory symptoms, some patients present other severe damage such as cardiovascular, renal and liver injury, and/or multiple organ failure, suggesting a spread of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in blood. Recent ultrasensitive polymerase chain reaction (PCR) technology now allows absolute quantification of nucleic acids in plasma. We intend to use the droplet-based digital PCR technology to obtain sensitive detection and precise quantification of plasma SARS-CoV-2 viral load (SARS-CoV-2 RNAemia) in hospitalized COVID-19 patients.

METHODS

Fifty-eight consecutive COVID-19 patients with pneumonia 8 to 12 days after onset of symptoms and 12 healthy controls were analyzed. Disease severity was categorized as mild to moderate in 17 patients, severe in 16, and critical in 26. Plasma SARS-CoV-2 RNAemia was quantified by droplet digital Crystal Digital PCR next-generation technology (Stilla Technologies, Villejuif, France).

RESULTS

Overall, SARS-CoV-2 RNAemia was detected in 43 (74.1%) patients. Prevalence of positive SARS-CoV-2 RNAemia correlated with disease severity, ranging from 53% in mild-to-moderate patients to 88% in critically ill patients (P = .036). Levels of SARS-CoV-2 RNAemia were associated with severity (P = .035). Among 9 patients who experienced clinical deterioration during follow-up, 8 had positive SARS-CoV-2 RNAemia at baseline, whereas only 1 critical patient with undetectable SARS-CoV-2 RNAemia at the time of analysis died at day 27.

CONCLUSION

SARS-CoV-2 RNAemia measured by droplet-based digital PCR constitutes a promising prognosis biomarker in COVID-19 patients.

摘要

背景

2019 年冠状病毒病（COVID-19）是一个全球性的公共卫生问题，已在全球范围内导致超过 662000 人死亡。虽然 COVID-19 的临床表现主要为呼吸道症状，但一些患者还出现其他严重损害，如心血管、肾脏和肝脏损伤和/或多器官衰竭，提示严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）在血液中的传播。最近的超灵敏聚合酶链反应（PCR）技术现在可以对血浆中的核酸进行绝对定量。我们打算使用基于液滴的数字 PCR 技术，对住院 COVID-19 患者的血浆 SARS-CoV-2 病毒载量（SARS-CoV-2 RNAemia）进行敏感检测和精确定量。

方法

分析了 58 例症状出现后 8 至 12 天的肺炎 COVID-19 患者和 12 例健康对照者。17 例患者为轻度至中度，16 例为重度，26 例为危重型。通过基于液滴的数字 Crystal Digital PCR 下一代技术（法国 Villejuif 的 Stilla Technologies）定量检测血浆 SARS-CoV-2 RNAemia。

结果

总体而言，43 例（74.1%）患者检测到 SARS-CoV-2 RNAemia。阳性 SARS-CoV-2 RNAemia 的检出率与疾病严重程度相关，从轻-中度患者的 53%到危重症患者的 88%不等（P=0.036）。SARS-CoV-2 RNAemia 水平与严重程度相关（P=0.035）。在 9 例随访期间病情恶化的患者中，8 例基线时 SARS-CoV-2 RNAemia 阳性，而在分析时 SARS-CoV-2 RNAemia 无法检测的 1 例危重症患者在第 27 天死亡。