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

描述（由申请方提供）：沙眼衣原体感染诊断的改善代表了一个关键的未满足的医疗需求。C.沙眼是人类的一种专性细胞内病原体，可导致失明、盆腔炎、不育，并且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头号细菌性传播疾病（STD）。其中许多感染是无症状的，导致了一场大规模的无声流行。目前的衣原体诊断主要基于核酸扩增测试（NAAT），这些测试缺乏不同NAAT之间的一致性，灵敏度不同（尽管特异性很高），价格昂贵，需要技术专长，需要数天才能获得结果（特别是批量处理），并且通常局限于主要的临床或参考实验室。他们也无法区分侵袭性性病淋巴肉芽肿（LGV）与非侵袭性菌株D至K，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）正计划开发一种廉价的基于微流体的C。沙眼，包括治愈测试，这将提供敏感和特异性的检测和菌株类型的区分，使临床决策能够在真实的时间。开发这种有效的床旁诊断可以导致广泛的筛查工作，以检测有症状和无症状的感染和适当的治疗，这不仅可以降低与这些感染直接相关的急性和慢性发病率，而且还可以降低与C相关的潜在附带风险。沙眼感染，包括HIV-1感染、宫颈癌和自身免疫驱动的关节炎。因此，该I期申请的具体目标是：1）开发用于从临床尿道和宫颈样品中提取基因组DNA的快速方法; 2）设计至少两个对C.沙眼衣原体在两个位点的扩增;和3）确定微流体诊断沙眼衣原体的灵敏度和特异性。与目前的NAAT相比，在SBIR II期，我们将：1）扩展我们的方法，从结膜、咽、尿液、溃疡、阴道和直肠样本中提取基因组DNA; 2）扩展我们的引物设计，根据参考和临床衣原体STD菌株的比较基因组学鉴定菌株类型（对于指导侵入性与非侵入性衣原体治疗至关重要）。沙眼衣原体菌株和流行病学研究）; 3）获得FDA批准的专利; 4）使用来自不同地理人群的衣原体性传播疾病风险的临床样本，设计我们的诊断与领先的商业诊断的头对头临床研究。鉴于Tim Read博士和Deborah Dean博士在开发靶基因引物对方面的基因组专业知识以及Dean博士的衣原体STD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C.沙眼公共卫生相关性：沙眼衣原体是全球性传播疾病的主要细菌。其中许多感染是无症状的，未被发现，这导致传播增加和盆腔炎、不孕症和异位妊娠的后遗症。目前的诊断需要广泛的技术专业知识，价格昂贵，需要几天的时间才能得出结果，导致失去后续行动。此外，需要更长时间治疗的更危险的菌株无法检测到。网络生物系统公司的目标是开发一种易于使用，成本效益高，一小时诊断C。还可以区分菌株类型以进行适当的治疗。

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.