Putra Henry Gotama, Surja Sem Samuel, Widowati Tria Asri, Ali Soegianto, Kaisar Maria Mardalena Martini
Undergraduate Study Program, School of Medicine and Health Sciences, Atma Jaya Catholic University of Indonesia, Jakarta, 14440 Indonesia.
Department of Parasitology, School of Medicine and Health Sciences, Atma Jaya Catholic University of Indonesia, Jakarta, 14440 Indonesia.
Virusdisease. 2024 Jun;35(2):293-301. doi: 10.1007/s13337-024-00870-1. Epub 2024 Jun 7.
Colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a potential and relatively simple rapid diagnostics method for COVID-19 detection. This study aims to evaluate and optimize the RT-LAMP performance on saliva specimens based on a commercially available kit.Modifications on an established protocol (Protocol A) were used, including Proteinase K supplementation (Protocol B); pre-treatment using nuclease-free water and proteinase K (Protocol C); Saliva cooling (Protocol D); saliva dilution after pre-treatment (Protocol E); lastly a combination of saliva cooling and dilution (Protocol F). Protocol performances were evaluated by comparing success rates (SR), diagnostic accuracy (DA), sensitivity, specificity, and predictive values. Additionally, a correlation between the Ct value by RT-qPCR and RT-LAMP performance was analyzed.. A total of 106 specimens were used in this study. Protocols B and C showed 100% unreadable results, therefore were paused. Protocol F showed the highest SR (87.65%) compared to other protocols, with a slight compromise to DA (81.69%), sensitivity (57.14%), specificity (97.67%), PPV (94.12%), and NPV (77.78%). In the sub-analysis of the low Ct value group (Ct < 30), Protocol F demonstrated a higher success rate (86.57%) compared to protocol A (64.18%); increased 3.08% sensitivity and 2.42% NPV; comparable DA; minor reduction in specificity (A = 100%; F = 97.67%) and PPV (A = 100%; F = 92.31%). A combination of saliva cooling-dilution substantially increased the tested kit's success rate, despite a slight decrease in specificity and PPV. Findings confirmed the saliva cooling-dilution procedure was beneficial to the test's SR, sensitivity, and NPV in the low Ct value group.
The online version contains supplementary material available at 10.1007/s13337-024-00870-1.
比色逆转录环介导等温扩增（RT-LAMP）是一种用于新冠病毒检测的有潜力且相对简单的快速诊断方法。本研究旨在基于市售试剂盒评估并优化唾液样本的RT-LAMP性能。对既定方案（方案A）进行了修改，包括补充蛋白酶K（方案B）；使用无核酸酶水和蛋白酶K进行预处理（方案C）；唾液冷却（方案D）；预处理后唾液稀释（方案E）；最后是唾液冷却和稀释相结合（方案F）。通过比较成功率（SR）、诊断准确性（DA）、敏感性、特异性和预测值来评估方案性能。此外，分析了RT-qPCR的Ct值与RT-LAMP性能之间的相关性。本研究共使用了106个样本。方案B和C显示100%结果不可读，因此暂停。与其他方案相比，方案F显示出最高的成功率（87.65%），但在诊断准确性（81.69%）、敏感性（57.14%）、特异性（97.67%）、阳性预测值（PPV，94.12%）和阴性预测值（NPV，77.78%）方面略有折衷。在低Ct值组（Ct＜30）的亚分析中，方案F显示出比方案A更高的成功率（86.57%对64.18%）；敏感性提高3.08%，阴性预测值提高2.42%；诊断准确性相当；特异性（方案A = 100%；方案F = 97.67%）和阳性预测值（方案A = 100%；方案F = 92.31%）略有降低。唾液冷却-稀释相结合显著提高了测试试剂盒的成功率，尽管特异性和阳性预测值略有下降。研究结果证实，唾液冷却-稀释程序对低Ct值组的测试成功率、敏感性和阴性预测值有益。
在线版本包含可在10.1007/s13337-024-00870-1获取的补充材料。
