Chang Chunyan, Li Lingling, Guo Yating, Ji Li, Tian Jinyue, Xu Shenglin, Zhang Xiuhong, Jiang Xinyi, Qiao Weizhen
Department of Laboratory Medicine, The Affiliated Wuxi People's Hospital of Nanjing Medical University, Wuxi People's Hospital, Wuxi Medical Center, Nanjing Medical University, Wuxi, China.
Blood Purification Center, The Affiliated Wuxi People's Hospital of Nanjing Medical University, Wuxi, China.
J Virol Methods. 2025 Jun;335:115151. doi: 10.1016/j.jviromet.2025.115151. Epub 2025 Apr 1.
Rapid, effective, and accurate detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is crucial. It is essential to control the spread of the virus and ensure accurate treatment for the disease. In the study, a total of 170 clinical specimens from 164 patients were collected and analyzed through digital PCR (dPCR) and real time quantitative reverse transcription PCR (RT-qPCR). The results showed an 86.41 % agreement between dPCR and RT-qPCR, with differences primarily noted in suspected cases. RT-qPCR exhibited a sensitivity of 84.78 %, specificity of 95.83 %, and accuracy of 86.42 %, which were comparatively lower than the 100 % accuracy of dPCR. Subsequently, we explored the potential correlation between these two methodologies based on Ct value groups. A strong negative correlation was observed between RT-qPCR and dPCR techniques in the Ct value group between 25 and 35, while the correlation was weakest in the Ct > 35 group. Moreover, the concordance rate for detecting the ORF1 (142/162) gene by RT-qPCR was lower compared to that of the N gene (149/162). Additionally, nucleic acid concentrations for ORF1 gene detection were lower than those for N gene detection in dPCR. In conclusion, this study shows that dPCR provides more reliable detection than RT-qPCR, especially for samples with low viral loads. Furthermore, dPCR effectively tracked changes in viral load during hospitalization, facilitating the diagnosis and treatment of COVID-19.
快速、有效且准确地检测严重急性呼吸综合征冠状病毒2（SARS-CoV-2）至关重要。这对于控制病毒传播以及确保对该疾病进行准确治疗必不可少。在这项研究中，共收集了164例患者的170份临床标本，并通过数字PCR（dPCR）和实时定量逆转录PCR（RT-qPCR）进行分析。结果显示，dPCR与RT-qPCR之间的一致性为86.41%，差异主要存在于疑似病例中。RT-qPCR的灵敏度为84.78%，特异性为95.83%，准确率为86.42%，相对低于dPCR的100%准确率。随后，我们基于Ct值分组探讨了这两种方法之间的潜在相关性。在Ct值为25至35的组中，RT-qPCR与dPCR技术之间观察到强烈的负相关，而在Ct>35的组中相关性最弱。此外，与N基因（149/162）相比，RT-qPCR检测ORF1（142/162）基因的一致性率较低。另外，在dPCR中，ORF1基因检测的核酸浓度低于N基因检测的核酸浓度。总之，本研究表明，dPCR比RT-qPCR提供更可靠的检测，尤其是对于病毒载量低的样本。此外，dPCR有效地跟踪了住院期间病毒载量的变化，有助于COVID-19的诊断和治疗。
