Low Cost Multiplexed DNA Diagnostic Sensors for Infectious Diseases

近2.6亿人感染了血吸虫病，其中90%的感染者在撒哈拉以南非洲地区发现。全世界约有32亿人面临疟疾的风险，其中许多人也在撒哈拉以南非洲。这两种疾病都在相同的农村地区流行，而且很难区分症状和提供正确的诊断和治疗。众所周知，错误的诊断会导致不必要的药物分配，从而增加产生耐药性的可能性。库珀教授开发了一种纸基多路DNA疟疾诊断设备，该设备利用新型热蜡打印将样品制备和微流控流相结合。该设备可以区分三种引起疟疾的疟原虫，现在已经在工厂使用完整的患者血液样本进行了测试。该项目的目的是开发一种新的基于血液的多重检测方法，可以从一滴血液中区分疟疾和血吸虫病，并识别抗药性微生物。这一高度敏感的诊断设备将使现场工作人员能够监测治疗结果，具体目标是：(1)提供正确的诊断；(2)防止局部治疗后再次出现感染；(3)通过确保有效和适当的治疗减少耐药性。现场测试将在非洲东部进行操作员双盲现场测试。样本的特征也将使用(I)纸质测试；(Ii)通过现场显微镜；(Iii)也将被储存(作为干血点)用于在格拉斯哥进行基于回顾的分子测试。将获得伦理许可。疟疾寄生虫(疟原虫)以红细胞为生命周期的一部分。因此，我们还将开发一种基于血液的检测方法，从一次挑选的血液中检测出DNA。该检测将再次配置为低成本的纸质检测，并将发展为双重检测，使疟疾和血吸虫病(血吸虫病)能够同时被检测到。这将指导在现场诊所对发烧患者进行适当的治疗。好处：该项目还与全球挑战基金的健康优先事项保持一致，该基金的目标是“解决疾病，加强卫生系统，并惠及世界上最脆弱的人群”。事实上，根据世卫组织的最新估计，2014年至少有2.58亿人需要治疗血吸虫病，据报道有6160万人需要治疗。90%的需要治疗的人生活在非洲。在儿童中，血吸虫病会导致贫血、生长发育迟缓和学习能力下降。治疗通常会逆转这些效果，但需要监测其疗效，以确定治愈和适应疗法。如果不治疗，慢性病会影响人们的工作能力，并可能导致死亡。疾病负担估计占450万DALY(残疾影响寿命年)。通过国家控制方案，例如在乌干达实施了十多年的大规模吡喹酮药物管理。发现了感染热点，其流行率、强度和相关发病率现在高于基线水平。

Nearly 260m people are infected with schistosomiasis, with >90% ofinfections found in sub-Saharan Africa. Worldwide ~3.2b people areat risk of malaria, many also in Sub-Saharan Africa. Both diseasesare endemic in the same rural locations and there is difficulty indifferentiating symptoms and informing correct diagnosis andtreatment. Incorrect diagnosis is known to lead to unnecessarydispensation of drugs leading to increased probability of drugresistance.Professor Cooper has developed a paper-based multiplexed DNAmalaria diagnostic device that exploits novel hot wax printing tointegrate sample preparation and microfluidic flows. The device candifferentiate between three species of malaria causing plasmodiumand has now been tested using whole patient blood samples in thelaboratory.The aim of this project is to develop a novel blood-based multiplextest that can, from a single drop of blood, distinguish between malariaand schistosomiasis and identify drug resistant microbes. This newhighly sensitive diagnostic device will enable field workers to monitortreatment outcomes, with the specific aims of(i) providing correct diagnosis;(ii) preventing re-emergence of infections following localtreatment;(iii) reducing resistance by ensuring effective and appropriatetreatment.Field-testing will be carried out operator double-blind field-testing inEast Africa. The samples will also be characterised using(i) the paper based test;(ii) through microscopy in-the-field(iii) will also be stored (as dried blood spots) for retrospectivelaboratory based molecular testing in Glasgow.Ethics permissions will be obtained.Additional Information: The schistosoma live in the hepatic portal veinand shed cells into the blood. Malaria parasites (Plasmodium sp.) livewith red blood cells as part of their lifecycle. We will therefore alsodevelop a blood based assay, where DNA will be detected from afingerpick of blood. The assay will again be configured as a low cost,paper based assay and will be developed as a duplex, enabling bothmalaria and bliharzia (schistosomiasis) to be detected at the sametime. This will guide appropriate treatment for febrile patientspresenting themselves at field clinics.Benefits:This project is also aligned with the Health priority of the GlobalChallenge Fund, which aims 'to tackle diseases, strengthen healthsystems and reach the worlds' most vulnerable'. Indeed, according tothe latest WHO estimates, at least 258 million people have requiredtreatment for schistosomiasis in 2014, with 61.6 million peoplereported to be treated. 90% of those requiring treatment live in Africa.In children, schistosomiasis causes anaemia, stunted growth andreduced ability to learn. Treatment usually reverses these effects, butits efficacy needs to be monitored to establish cure and adapttherapy. If untreated, chronic disease affects people's ability to workand can result in death. The disease burden has been estimated toaccount for 4.5 million DALYs (disability affected life years).Mass drug administration of praziquantel has been delivered throughNational Control Programmes, e.g. in Uganda for over a decade.Hotspots of infection have been detected, where prevalence,intensity and associated morbidity are now higher than they were atbaseline.