Hu Junjie, Xu Shanshan, Zeng Zeng, Gong Wei, Xu Weihua, Ma Zhichao, Fu Shengmiao, Li Linhai, Xiao Bin, Chen Xinping
Department of clinical Laboratory, Affiliated Cancer Hospital of Hainan Medical University, Hainan Cancer Hospital, Haikou, Hainan, People's Republic of China.
The First Clinical School of Hainan Medical University, Haihou, Hainan, People's Republic of China.
Microbiol Spectr. 2025 Sep 22:e0059225. doi: 10.1128/spectrum.00592-25.
) is a gram-negative bacterium found in soil and surface water. It is also the pathogen that causes melioidosis disease in humans and animals. This study aimed to obtain the whole genome sequence of the endemic strain of in Hainan, using third-generation sequencing (TGS) technology, and elucidate the genome structure, function, and genetic evolution. Additionally, the study aimed to achieve rapid and specific identification of these endemic strains using specific high-sensitivity enzymatic reporter unlocking (SHERLOCK) detection technology, providing a new strategy for the early diagnosis of melioidosis. Utilizing the PacBio platform for TGS technology, we completed whole genome sequencing of 16 strains from Hainan. High-precision and complete genome sequences were obtained through quality control and genome assembly of the sequencing data. Additionally, we established a nucleic acid detection technology platform based on SHERLOCK, which could be completed from nucleic acid extraction to result reading within 1-2 hours, demonstrating good sensitivity and specificity (both are 100%). The lateral chromatography strip method does not require special equipment and holds promise as an immediate screening method for the early diagnosis of melioidosis.
Melioidosis is a highly pathogenic infectious disease caused by a gram-negative bacterium of (). The traditional gold standard for diagnosing melioidosis is still isolation and culture from clinical samples. Although this method has high specificity, it has low sensitivity and is time-consuming, which often leads to misdiagnosis or missed diagnosis of melioidosis, affecting subsequent treatment. In this study, recombinase polymerase amplification technology and clustered regularly interspaced short palindromic repeats/Cas13a technology were combined to establish the Specific High-sensitivity Enzymatic Reporter Unlocking detection technology, which can achieve rapid and accurate identification of , providing a new method for the early diagnosis of melioidosis.
）是一种存在于土壤和地表水中的革兰氏阴性菌。它也是导致人类和动物类鼻疽病的病原体。本研究旨在利用第三代测序（TGS）技术获取海南地方性菌株的全基因组序列，并阐明其基因组结构、功能和遗传进化。此外，该研究旨在利用特异性高灵敏度酶促报告分子解锁（SHERLOCK）检测技术实现对这些地方性菌株的快速和特异性鉴定，为类鼻疽病的早期诊断提供新策略。利用PacBio平台进行TGS技术，我们完成了来自海南的16株菌株的全基因组测序。通过对测序数据的质量控制和基因组组装获得了高精度和完整的基因组序列。此外，我们建立了基于SHERLOCK的核酸检测技术平台，该平台可在1 - 2小时内完成从核酸提取到结果读取，显示出良好的灵敏度和特异性（均为100%）。侧向层析条法不需要特殊设备，有望作为类鼻疽病早期诊断的即时筛查方法。
类鼻疽病是由（）革兰氏阴性菌引起的高致病性传染病。诊断类鼻疽病的传统金标准仍然是从临床样本中分离和培养。虽然这种方法具有高特异性，但灵敏度低且耗时，这常常导致类鼻疽病的误诊或漏诊，影响后续治疗。在本研究中，将重组酶聚合酶扩增技术和规律成簇间隔短回文重复序列/Cas13a技术相结合，建立了特异性高灵敏度酶促报告分子解锁检测技术，可实现对（）的快速准确鉴定，为类鼻疽病的早期诊断提供了新方法。
