Giner Inc., 89 Rumford Avenue, Newton, MA, 02466, United States.
J. Craig Venter Institute (JCVI), 4120 Capricorn Lane La Jolla, CA, 92037, United States.
Biosens Bioelectron. 2023 Feb 15;222:114986. doi: 10.1016/j.bios.2022.114986. Epub 2022 Dec 2.
Fungal infections are a rapidly increasing public health problem due to their high morbidity and mortality rates, especially in populations with compromised immune systems. Rapid and accurate diagnosis of these diseases is, therefore, necessary to improve the prognosis of afflicted patients. Unfortunately, current clinical chemistry practice relies on lengthy culturing methods that are insufficient to meet the fast turnaround requirements. Here we present a cost-effective and robust nucleic acid sensor that can identify the presence of histoplasmosis causing fungal genes, in whole blood or bronchoalveolar lavage (BAL) samples, far earlier than current methods. Our novel assay involves the hybridization of target gene sequences with immobilized nucleic acid probes, allowing direct, label-free detection of Hcp100, CBP1, and M antigen genes through electrochemical analysis. The resultant current is attributed to the presence of fungal targets in the sample solution. The assay provides ultra-sensitive detection of DNA molecules with a limit of detection (LOD) values down to 100 aM, sufficient to meet the clinical diagnostic need. In addition, the turnaround time for the sample to result is less than 90 min compared to the current clinical procedure's turnaround time of 3-4 weeks.
真菌感染是一个迅速增长的公共卫生问题，因为它们具有很高的发病率和死亡率，尤其是在免疫系统受损的人群中。因此，快速准确地诊断这些疾病对于改善受影响患者的预后是必要的。不幸的是，目前的临床化学实践依赖于冗长的培养方法，这些方法不足以满足快速周转的要求。在这里，我们提出了一种具有成本效益的、强大的核酸传感器，它可以在全血或支气管肺泡灌洗液（BAL）样本中比当前方法更早地识别引起组织胞浆菌病的真菌基因的存在。我们的新测定法涉及与固定化核酸探针杂交目标基因序列，通过电化学分析直接、无标记地检测 Hcp100、CBP1 和 M 抗原基因。所得电流归因于样品溶液中真菌靶标的存在。该测定法对 DNA 分子具有超灵敏的检测能力，检测限（LOD）值低至 100 aM，足以满足临床诊断需求。此外，与当前临床程序的 3-4 周周转时间相比，样本的周转时间不到 90 分钟。
