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Enhancement of Lateral Flow Assay for the Detection of Whole Viral Particle and Chlamydial Elementary Bodies

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Abstract
Chlamydia trachomatis accounts for 3.6% of blindness worldwide, and is the leading cause of bacterial-induced blindness in the world. With the subtle initial presentation of the disease and the difficulty in clearing the infection without the aid of antibiotics, C. trachomatis can spread rapidly following introduction into a population. This problem is further compounded in resource limited areas due to the lack of trained personnel (i.e. Medical Doctors, Nurses), equipment, and finances to test and treat large portions of the population. A testing method that is both cheap and easy to interpret is necessary. Lateral flow assays (LFA) have been used for years to evaluate pregnancy status in the developed world, and their low cost, ease of use and disposable nature make them a worthwhile candidate, but the current use of visual reporters (i.e. gold or latex nanoparticles) does not allow for adequate sensitivity for true clinical use. Fluorescent reporters, particularly fluorescent nanoparticles, would lower the limit of detection (LOD) and allow for the detection of acute and subclinical infections, which would allow for an effective and objective screening method for trachoma and many other diseases. An effective, rapid, and disposable test would allow for mass screening to be implemented which, in turn, would allow for rapid and targeted treatment. The results in this study show that the use of fluorescent-based reporters greatly improve the LOD of the LFA, with both FITC and RuSNP reporters showing a reduction in the LOD by 1 and 2.5 logs respectively when compared to traditional colorimetric reporters. This substantial improvement in the LOD of the LFA allows for the tests to be used to detect relevant levels of viral pathogens. A similar improvement in the LOD was seen when using FITC-labeled antibodies which improved the sensitivity of LFAs with regards to the detection of C. trachomatis. The use of fluorescent-based reporters in LFAs greatly improves the LOD for both viruses and bacteria, allowing for their detection at clinically relevant levels.
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2014-02
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