Improved Chlamydia Test Using Aptamers on a Portable POC Microfluidic Platform

在便携式 POC 微流控平台上使用适体改进衣原体测试

• 批准号: 8698872
• 负责人: Vladimir Gilman
• 金额: $ 1.27万
• 依托单位: VIVONICS, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8698872
• 关键词: Improved; Chlamydia; Test; Using; Aptamers

DESCRIPTION (provided by applicant): The fundamental goal of this program is to develop a point of care high fidelity, aptamer based assay for genital Chlamydia infections that allows fast and quantitative determination of levels of infection with different sero-types of Chlamydia trachomatis. The ultimate goal is to have this assay be integrated into an inexpensive portable microfluidics device that can be used in any physician's office. This device will provide vital information on important health aspects of some underserved populations. The key element is a family of serotype specific DNA aptamers that exhibit very high sensitivity and specificity (99,999 %) of detection of C. trachomatis. The DNA aptamers act as an enhanced replacement for the antibodies and specifically react with the target microorganisms in a microfluidic version of sandwich ELISA realized in a portable hand held device recently developed by one of the team members, Claros Diagnostics (Cambridge MA). A two phase program is proposed. The Phase I culminates in the validation of the aptamer assay for Chlamydia trachomatis in the International STD, Respiratory, and Biothreat Research Laboratory of Johns Hopkins University (Baltimore, MD). The Phase II of the program will focus on bringing together the detection biochemistry and instrumental grid in a new point of care (physician's office) assay for Chlamydia trachomatis. The preliminary data accumulated by the proposal team encompass extensive experience and profound knowledge of clinical and societal aspects of C. trachomatis infections brought by the group of Professor Charlotte A. Gaydos, Johns Hopkins University. Claros Diagnostics' microfluidics detection platform realized as a small device is capable of dealing with microscale bio-samples and allows rapid reproducible handling of microliter liquid volumes. The novel aptamer selection technology has been developed by Infoscitex Corporation and has been successfully applied to produce high quality DNA aptamers for the US Department of Defense applications, such as detection of anthrax toxin and infectious prions. PUBLIC HEALTH RELEVANCE: Currently, the most sensitive diagnosis of genital chlamydiosis employs quantitative real-time polymerase chain reaction qRT-PCR. As recently reported, the major outer membrane protein (omp-1) -gene and a L- serovar-specific region of the polymorphic protein H (pmp-H) -gene could be used to detect C. trachomatis. The detection limit of each real-time PCR was 50 genome copies per reaction (Schaeffer, 2008). However, difficulties in distinguishing some of the C. trachomatis sero-variants by this method were acknowledged by the authors. If implemented on a large scale, the costs of RT-PCR diagnosis for C. trachomatis and time to diagnosis may, unfortunately, become prohibitive for the underserved populations. Therefore, a rapid and affordable point of care analysis of patient-derived specimens can revolutionize the field of diagnosis of genital chlamydial infections. Our technology will provide physicians with an inexpensive simple assay and unambiguous readout that will suggest, or rule out, the necessity of further treatment right in the physician's office. In summary, the assay-device combination proposed here will reach the underserved populations who can be much more fully protected when enabled with point of care diagnosis of genital chlamydial infections.

描述（由申请方提供）：该项目的基本目标是开发一种用于生殖器衣原体感染的基于适体的高保真床旁检测方法，该方法可以快速定量测定不同血清型沙眼衣原体的感染水平。最终目标是将这种检测方法集成到一种廉价的便携式微流体设备中，可以在任何医生的办公室中使用。该设备将提供关于某些得不到充分服务的人口的重要健康方面的重要信息。关键元件是血清型特异性DNA适体家族，其表现出非常高的检测C的灵敏度和特异性（99，999%）。沙眼DNA适体作为抗体的增强替代物，并在最近由团队成员之一Claros Diagnostics（剑桥MA）开发的便携式手持设备中实现的夹心ELISA的微流体版本中与靶微生物特异性反应。提出了两个阶段的计划。I期研究的高潮是在约翰霍普金斯大学（巴尔的摩，MD）的国际STD、呼吸道和生物威胁研究实验室验证沙眼衣原体的适体检测。该计划的第二阶段将重点放在将检测生物化学和仪器网格结合在一起，用于沙眼衣原体的新的护理点（医生办公室）检测。提案团队积累的初步数据包括C的临床和社会方面的广泛经验和深刻知识。由夏洛特教授A.同性恋，约翰霍普金斯大学。Claros Diagnostics的微流体检测平台实现为小型设备，能够处理微量生物样品，并允许快速再现处理微升液体体积。新的适体选择技术已由Infoscitex公司开发，并已成功应用于生产高质量的DNA适体，用于美国国防部的应用，如炭疽毒素和传染性朊病毒的检测。 公共卫生相关性：目前，生殖器衣原体病最敏感的诊断采用定量实时聚合酶链反应qRT-PCR。最近报道，主要外膜蛋白（omp-1）基因和L-血清型特异性多态性蛋白H（pmp-H）基因可用于检测C。沙眼每个实时PCR的检测限为每个反应50个基因组拷贝（Schaeffer，2008）。然而，在区分某些C.沙眼衣原体血清变异体通过该方法被作者认可。如果大规模实施，RT-PCR诊断C。不幸的是，沙眼和诊断时间可能会使服务不足的人群望而却步。因此，对患者来源的标本进行快速且经济实惠的护理点分析可以彻底改变生殖器衣原体感染的诊断领域。我们的技术将为医生提供一个便宜的简单的测定和明确的读数，将建议，或排除，进一步治疗的必要性，在医生的办公室。总之，本文提出的检测-器械组合将覆盖服务不足的人群，当能够进行生殖器衣原体感染的护理点诊断时，这些人群可以得到更充分的保护。