Improving RT-PCR detection from frozen, unpreserved first-void clinical samples
APPLICATION NOTE SKU 44XXX
Key Advantages
> Capture and concentrate Zika virus from urine to boost assay sensitivity and eliminate the need for invasive blood collection.
> Overcome the sensitivity challenges in frozen storage conditions by minimizing the impact of sample degradation through virus enrichment.
> Circumvent the low input sample volume restrictions of commercial extraction methods by capturing more virions from larger volumes upstream.
> Easily integrate into existing workflows using sample collection devices, commercial RNA extraction methods and RT-PCR assays.
Introduction
Increasing geographic distribution of mosquito vectors and subsequent transmission by international travel has created a need for a technology that is capable of accurately and reliably detecting low-concentration viral pathogens, like Zika, in biological samples. Non-invasive collection methods, such as urine sampling are ideal, but they are limited by low molecular assay sensitivity in urine samples. This sensitivity issue is further exacerbated when collected samples need to be stored for longer periods of time or transported prior to testing. First-void urine improves the sensitivity of molecular assays and offers an alternative to serum sampling. However, testing of urine samples that are stored for extended periods of time (> 1 year) in the absence of a preservative can result in false-negative results.
Nanotrap particles can capture, concentrate, and preserve viral pathogens from biological fluids upstream of a range of analytical methods, including immunoassays and quantitative reverse-transcription polymerase chain reaction (qRT-PCR).
In this study, Nanotrap particles were evaluated for their ability to significantly improve the RT-PCR detection of Zika virus in frozen, archived clinical first-void urine samples. This work was performed in collaboration with Escuela Superior Politecnica del Litoral (ESPOL), Ghent University, Eucagen LLC., and Novosanis.
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