Arkell Paul, Mairiang Dumrong, Songjaeng Adisak, Malpartida-Cardenas Kenny, Hill-Cawthorne Kerri, Avirutnan Panisadee, Georgiou Pantelis, Holmes Alison, Rodriguez-Manzano Jesus
Centre for Antimicrobial Optimisation, Department of Infectious Disease, Imperial College London, Hammersmith Hospital, London, United Kingdom.
Siriraj Center of Research Excellence in Dengue and Emerging Pathogens (SiCORE-Dengue), Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
PLOS Glob Public Health. 2023 Aug 8;3(8):e0002169. doi: 10.1371/journal.pgph.0002169. eCollection 2023.
Dengue is a mosquito-borne disease caused by dengue virus (DENV) serotypes 1-4 which affects 100-400 million adults and children each year. Reverse-transcriptase (RT) quantitative polymerase chain reaction (qPCR) assays are the current gold-standard in diagnosis and serotyping of infections, but their use in low-middle income countries (LMICs) has been limited by laboratory infrastructure requirements. Loop-mediated isothermal amplification (LAMP) assays do not require thermocycling equipment and therefore could potentially be deployed outside laboratories and/or miniaturised. This scoping literature review aimed to describe the analytical and diagnostic performance characteristics of previously developed serotype-specific dengue RT-LAMP assays and evaluate potential for use in portable molecular diagnostic devices. A literature search in Medline was conducted. Studies were included if they were listed before 4th May 2022 (no prior time limit set) and described the development of any serotype-specific DENV RT-LAMP assay ('original assays') or described the further evaluation, adaption or implementation of these assays. Technical features, analytical and diagnostic performance characteristics were collected for each assay. Eight original assays were identified. These were heterogenous in design and reporting. Assays' lower limit of detection (LLOD) and linear range of quantification were comparable to RT-qPCR (with lowest reported values 2.2x101 and 1.98x102 copies/ml, respectively, for studies which quantified target RNA copies) and analytical specificity was high. When evaluated, diagnostic performance was also high, though reference diagnostic criteria varied widely, prohibiting comparison between assays. Fourteen studies using previously described assays were identified, including those where reagents were lyophilised or 'printed' into microfluidic channels and where several novel detection methods were used. Serotype-specific DENV RT-LAMP assays are high-performing and have potential to be used in portable molecular diagnostic devices if they can be integrated with sample extraction and detection methods. Standardised reporting of assay validation and diagnostic accuracy studies would be beneficial.
登革热是一种由1 - 4型登革病毒（DENV）引起的蚊媒疾病，每年影响1亿至4亿成年人和儿童。逆转录酶（RT）定量聚合酶链反应（qPCR）检测是目前感染诊断和血清分型的金标准，但它们在低收入和中等收入国家（LMICs）的使用受到实验室基础设施要求的限制。环介导等温扩增（LAMP）检测不需要热循环设备，因此有可能在实验室外部署和/或小型化。本综述性文献旨在描述先前开发的血清型特异性登革热RT - LAMP检测的分析和诊断性能特征，并评估其在便携式分子诊断设备中的应用潜力。我们在Medline中进行了文献检索。如果研究在2022年5月4日之前列出（未设置先前的时间限制），并且描述了任何血清型特异性DENV RT - LAMP检测的开发（“原始检测”），或者描述了这些检测的进一步评估、改编或实施，则纳入研究。收集了每个检测的技术特征、分析和诊断性能特征。共鉴定出8种原始检测。这些检测在设计和报告方面存在差异。检测的最低检测限（LLOD）和定量线性范围与RT - qPCR相当（对于定量目标RNA拷贝的研究，报告的最低值分别为2.2×101和1.98×102拷贝/毫升），分析特异性较高。在进行评估时，诊断性能也较高，尽管参考诊断标准差异很大，无法进行检测之间的比较。我们还鉴定了14项使用先前描述的检测的研究，包括试剂冻干或“打印”到微流控通道中的研究，以及使用了几种新型检测方法的研究。血清型特异性DENV RT - LAMP检测性能良好，如果能够与样本提取和检测方法集成，有可能用于便携式分子诊断设备。对检测验证和诊断准确性研究进行标准化报告将是有益的。
