Integrating CRISPR-Cas diagnostics into organic electronics for point-of-care genotyping

庆大霉素是一种氨基糖苷类抗生素，常用于治疗新生儿感染。在英国，每年约有90，000名婴儿在新生儿重症监护室接受庆大霉素治疗（全球700万）。国家和国际指南强调快速抗生素给药的重要性，对于疑似败血症的病例，最好在一小时内给予抗生素。在高剂量或长期治疗方案中，氨基糖苷类会引起耳毒性，表现为听力或平衡障碍。然而，某些个体具有氨基糖苷类诱导的耳毒性（AIO）的遗传易感性，因此仅单次剂量就会导致严重和不可逆的听力损失。这些人在线粒体RNR 1基因（m.1555AG）中有一个变体，在一般人群中的患病率为0.2%（1/500）。因此，一种从口腔拭子样本中快速（在几分钟内）确定患者遗传状态的方法将具有重要的临床价值，通过确定是否应使用替代抗生素来避免AIO.This项目旨在开发一种生物电子设备来检测人类口腔样本中的m.1555变体。目前，患者的遗传状态是通过基于PCR的方法确定的，该方法需要复杂的实验室设备、昂贵的试剂、专业人员，并且对于这种临床情况来说不够快速。拟议的设备将生化检测变异的存在，并提供一个易于解释的数字读出。这样的电子输出然后可以被传输到患者的电子记录，而不需要手动解释和转录结果，从而节省时间并降低人为错误的风险。

Gentamicin is an aminoglycoside antibiotic commonly used to treat neonatal infections. Each year in the UK, approximately 90,000 babies are administered gentamicin on Neonatal Intensive Care Units (7 million worldwide). National and international guidelines stress the importance of rapid antibiotic administration, ideally within an hour for cases of suspected sepsis. In high doses, or with protracted regimens, aminoglycosides causes ototoxicity that can manifest as hearing or balance impairment. However, certain individuals have a genetic predisposition to aminoglycoside induced ototoxicity (AIO), whereby just a single dose causes profound and irreversible hearing loss. These individuals have a variant in the mitochondrial RNR1 gene (m.1555AG), which has a prevalence of 0.2% (1 in 500) in the general population. A method to rapidly (within minutes) determine a patient's genetic status from a buccal swab sample would therefore be of significant clinical value, by establishing if alternative antibiotics should be used instead to avoid AIO.This project aims to develop a bioelectronic device to detect the m.1555 variant from human buccal samples. Currently, a patient's genetic status is determined by PCR-based methods that require complex laboratory equipment, expensive reagents, specialist personnel, and are insufficiently rapid for this clinical scenario. The proposed device will biochemically detect the presence of the variant and deliver an easily interpretable digital read-out. Such an electronic output can then be transferred to a patient's electronic records, without the need for manual interpretation and transcription of the results, thus saving time and reducing the risk of human error.