Development, assessment, an implementation of a dual isothermal point-of care molecular diagnostic for both FGS and HPV

FGS 和 HPV 双等温护理点分子诊断的开发、评估和实施

• 批准号: 2881970
• 金额: --
• 依托单位: London School of Hygiene & Tropical Medicine
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2881970
• 关键词: Development; assessment; implementation; dual; isothermal

FGS is a gynaecological condition resulting from infection with the parasite Schistosoma haematobium and is thought to affect over 40 million women living across sub-Saharan Africa. In addition to causing symptoms such as bloody urine, severe cramping, and infertility, S. haematobium infection is associated with heightened risk of STI's and is a Grade 1 carcinogen. HPV is group of viruses known to cause genital warts and is often a precursor to cervical cancer. HPV is a global pathogen, and high-risk HPV strains cause more than 90% of all cervical cancer cases. A major hurdle in successful diagnosis and treatment in for Least Developed Countries (LDC's) is the lack of accessible diagnostics. Due to the fact that both HPV and FGS can be detected through gynaecological samples, this project aims to provide an integrated diagnostic that can be implemented in low resource settings to support timely treatment of marginalised women.Project Plan1. To review current literature to identify isothermal molecular assays that have been developed and used for the diagnosis of FGS and HPV. These assays will also be reviewed in terms of their applicability in low resource settings. 2. Suitable targets and primer-probe sets for RPA and LAMP assay development will be selected or redesigned (if needed) using genetic data from open-access databases. Online resources, including BLAST and OligoAnalyzer, will be used to in silico test the specificity of the assays followed by laboratory analytical development and testing. For HPV, a general HPV target and a high-risk HPV target would both be investigated.3. Assays will be tested and optimised as singleplex reactions before being optimised in a multiplex.4. An internal control target (human DNA) will be investigated and aimed to be incorporated into the assay for quality control purposes.5. Analytical testing will be done using synthetic DNA, gDNA from reference samples, and also available clinical samples for both S. haematobium and HPV. All molecular tests will be validated against qPCR reference tests.6. The assays we be evaluated in terms of their fit to the REASSURED criteria set out for diagnostics from the WHO for diagnostics for LDC's.7. Final stage assessment of the assay performance and suitability will be performed in a S. haematobium / HPV co-endemic setting in Sub-Saharan Africa. A comparison between self-swab and clinical swab samples will also be made to help inform the acceptability of the sampling procedure within the target population. Considering cultural and social attitudes is essential in relation to the most accurate diagnosis8. As well as descriptive statistics, differences in diagnostic capabilities between qPCR and the developed multiplex assay would be carried out using latent class analysis. The project will develop interdisciplinary skills including molecular biology, chemistry, field work, epidemiological analysis, and statistical analysis. Knowledge of physiology and pathology, skills identified as vulnerable by the UKRI, will also be developed.There is emphasis on intercollegiate collaboration, with significant portions of the project being undertaken at each institution due the bespoke expertise of each partner. Additionally, existing connections with commercial diagnostic companies and non-for-profit organisations that are dedicated to supporting work towards the provision of accessible diagnostics to marginalized communities, will guide this research.The project considers the MRC LID themes of global health and infectious disease, which are essential to the MRC strategy of improving human health and economic prosperity for everyone. Schistosomiasis is a neglected tropical disease and is most prevalent in LDC's. HPV is present globally, but access to screening and vaccination is limited in LDCs.

FGS是一种由寄生虫埃及血吸虫感染引起的妇科疾病，被认为影响了生活在撒哈拉以南非洲的4000多万妇女。除了引起血尿、严重痉挛和不孕等症状外，S。嗜血杆菌感染与STI的高风险相关，是1级致癌物。HPV是一组已知会导致生殖器疣的病毒，通常是宫颈癌的前兆。HPV是一种全球性病原体，90%以上的宫颈癌病例是由高危HPV毒株引起的。最不发达国家在成功诊断和治疗方面的一个主要障碍是缺乏可获得的诊断。由于HPV和FGS都可以通过妇科样本检测出来，该项目旨在提供一种可以在资源匮乏的环境中实施的综合诊断，以支持边缘化妇女的及时治疗。回顾当前文献，以确定已开发并用于诊断FGS和HPV的等温分子检测。还将审查这些测定法在低资源环境中的适用性。2.将使用来自开放访问数据库的遗传数据选择或重新设计（如需要）用于RPA和LAMP测定开发的合适靶标和引物-探针组。在线资源（包括BLAST和OligoAnalyzer）将用于计算机模拟检测试验的特异性，然后进行实验室分析开发和检测。对于HPV，一般HPV靶点和高危HPV靶点都将进行研究。在多路优化之前，将检测试剂盒作为单路反应进行测试和优化。4.将研究内部对照靶标（人DNA），并旨在将其纳入测定中用于质量控制样品。将使用合成DNA、参比样品的gDNA以及两种S的可用临床样品进行分析检测。嗜血杆菌和HPV。所有分子检测均将根据qPCR参考检测进行验证。6.我们将根据其对WHO为诊断LDC而制定的REASSURED诊断标准的适合性对测定进行评价。试验性能和适用性的最终阶段评估将在S.撒哈拉以南非洲的埃及伊蚊/ HPV共同流行环境。还将对自拭子和临床拭子样本进行比较，以帮助了解采样程序在目标人群中的可接受性。考虑文化和社会态度对于最准确的诊断至关重要8。除了描述性统计，qPCR和开发的多重检测试剂盒之间的诊断能力差异将使用潜在类别分析进行。该项目将培养跨学科技能，包括分子生物学，化学，实地工作，流行病学分析和统计分析。生理学和病理学的知识，被英国研究所确定为脆弱的技能，也将得到发展。重点是校际合作，由于每个合作伙伴的定制专业知识，每个机构都将承担项目的重要部分。此外，与商业诊断公司和非营利组织的现有联系，致力于支持向边缘化社区提供无障碍诊断的工作，将指导这项研究。该项目考虑了MRC LID的全球健康和传染病主题，这对MRC改善人类健康和每个人的经济繁荣战略至关重要。血吸虫病是一种被忽视的热带疾病，在最不发达国家最为流行。HPV在全球都存在，但在最不发达国家，筛查和疫苗接种的机会有限。