A Rapid Point-of-care Diagnostic for C.trachomitis STDs

描述（由申请人提供）：改进沙眼衣原体感染的诊断是一个关键的未满足的医疗需求。沙眼原体是一种专性的人类细胞内病原体，可导致失明、盆腔炎、不孕症，是人类免疫缺陷病毒（HIV）传播的重要辅助因子。与这种病原体相关的感染率使其成为全球排名第一的细菌性传播疾病（STD）。这些感染中有许多是无症状的，造成了大规模的无声流行。目前的衣原体诊断主要基于核酸扩增试验（NAAT），不同NAAT之间缺乏一致性，灵敏度不同（尽管特异性很高），价格昂贵，需要专业技术，需要数天才能得到结果（特别是批量处理），并且通常仅限于主要临床或参考实验室。它们也无法区分侵袭性性病淋巴肉芽肿（LGV）与非侵袭性D - K菌株，前者需要数周的抗生素治疗才能根除。网络生物系统公司（NetBio）正在提议开发一种廉价的基于微流体技术的沙眼原体诊断方法，包括治愈试验，这将提供对菌株类型的敏感和特异性检测和区分，从而使临床决策能够实时做出。开发这样一种有效的即时诊断方法可能会导致广泛的筛查工作，以发现有症状和无症状的感染，并进行适当的治疗，这不仅可以减少与这些感染直接相关的急性和慢性发病率，还可以减少与沙眼衣原体感染相关的潜在附带风险，包括HIV-1感染、宫颈癌和自身免疫性关节炎。因此，该I期申请的具体目标是：1)开发一种快速提取临床尿道和宫颈样本基因组DNA的方法；2)设计至少两对对沙眼衣原体两个位点扩增具有敏感性和特异性的引物；3)与目前的NAAT相比，确定微流体诊断沙眼衣原体的敏感性和特异性。在SBIR二期研究中，我们将：1)扩展我们从结膜、咽、尿液、溃疡、阴道和直肠样本中提取基因组DNA的方法；2)扩展引物设计，根据参考和临床衣原体性病菌株的比较基因组学确定菌株类型（对于指导侵袭性沙眼衣原体菌株与非侵袭性沙眼衣原体菌株的治疗和流行病学研究至关重要）；3)获得FDA批准的专利；4)设计一项针对我们的诊断与领先的商业诊断的面对面临床研究，使用来自衣原体性传播疾病风险的不同地理人群的临床样本。鉴于基因组学博士的专业知识。Tim Read和Deborah Dean开发目标基因引物对，以及Dean博士的衣原体性病专业知识，该应用程序提供了一个独特的合作机会，最终获得沙眼衣原体的快速即时诊断。公共卫生相关性：沙眼衣原体是世界范围内性传播疾病的主要细菌病因。许多这些感染是无症状的，未被发现，这导致传播增加和盆腔炎的后遗症，不孕症和异位妊娠。目前的诊断需要广泛的专业技术知识，价格昂贵，需要数天才能得出结果，导致失去随访。此外，需要更长时间治疗的更危险的菌株无法被发现。Network Biosystems的目标是开发一种易于使用、具有成本效益的沙眼衣原体一小时诊断方法，该方法还可以区分菌株类型，以便进行适当的治疗。

DESCRIPTION (provided by applicant): Improved diagnoses of Chlamydia trachomatis infections represent a critical unmet medical need. C. trachomatis is an obligate intracellular pathogen of humans that causes blindness, pelvic inflammatory disease, infertility, and is an important cofactor in transmission of human immunodeficiency virus (HIV). Infection rates associated with this pathogen make it the number one bacterial sexually transmitted disease (STD) worldwide. Many of these infections are asymptomatic, contributing to a large, silent epidemic. Current chlamydial diagnostics are primarily based on nucleic acid amplification tests (NAAT) that lack concordance across the different NAATs, vary in sensitivity (although specificity is high), are expensive, require technical expertise, take days for results (especially with batch processing), and are usually confined to major clinical or reference laboratories. They are also unable to differentiate between invasive lymphogranuloma venereum (LGV) versus non-invasive strains D to K, the former of which require weeks of antibiotic therapy for eradication. Network Biosystems (NetBio) is proposing to develop an inexpensive microfluidics-based diagnostic for C. trachomatis, including test-of-cure, that will provide sensitive and specific detection and discrimination of strain types to enable clinical decisions to be made in real time. Development of such an effective point-of-care diagnostic could result in widespread screening efforts for detection of symptomatic and asymptomatic infections and appropriate treatment that would not only reduce the acute and chronic morbidity that is directly associated with these infections but could also reduce potential collateral risks associated with C. trachomatis infections, including HIV-1 infection, cervical cancer and autoimmune-driven arthritis. The Specific Aims, then, of this Phase I application are to: 1) Develop a rapid method for genomic DNA extraction from clinical urethral and cervical samples; 2) Design at least two primer pairs that are sensitive and specific for C. trachomatis amplification at two loci; and 3) Determine the sensitivity and specificity of the microfluidics diagnostic for C. trachomatis compared to a current NAAT. In the SBIR Phase II, we will: 1) Expand our methods to extract genomic DNA from conjunctival, pharyngeal, urine, ulcer, vaginal and rectal samples; 2) Expand our primer design to identify strain types based on comparative genomics of reference and clinical chlamydial STD strains (essential for directing therapy for invasive versus non-invasive C. trachomatis strains and for epidemiologic studies); 3) Secure patents for FDA approval; 4) Design a head-to-head clinical study of our diagnostic against leading commercial diagnostics using clinical samples from diverse geographic populations at risk for chlamydial STDs. Given the genomic expertise of Drs. Tim Read and Deborah Dean for developing target gene primer pairs and the chlamydial STD expertise of Dr. Dean, the application provides a unique collaborative opportunity to finally obtain a rapid point-of-care diagnostic for C. trachomatis. PUBLIC HEALTH RELEVANCE: Chlamydia trachomatis is the leading bacterial cause of sexually transmitted diseases worldwide. Many of these infections are asymptomatic and go undetected, which leads to increased transmission and the sequelae of pelvic inflammatory disease, infertility and ectopic pregnancy. Current diagnostics require extensive technical expertise, are expensive and take days for results, leading to loss of follow up. Additionally, more dangerous strains that require longer treatment cannot be detected. Network Biosystems aims to develop an easy to use, cost effective, one hour diagnostic for C. trachomatis that also differentiates strains types for appropriate treatment.