Oligonucleotide Array-in-Well Platform for Detection and Genotyping Human Adenoviruses by Utilizing Upconverting Phosphor Label Technology

Abstract

We have developed a robust array-in-well test platform based on an oligonucleotide array, combining advantages of simple instrumentation and new upconverting phosphor reporter technology. Upconverting inorganic lanthanide phosphors have a unique property of photoluminescence emission at visible wavelengths under near-infrared excitation. No autofluorescence is produced from the sample or support material, enabling a highly sensitive assay. In this study, the assay is performed in standard 96-well microtiter plates, making the technique easily adaptable to high-throughput analysis. The oligonucleotide array-in-well assay is employed to detect a selection of ten common adenovirus genotypes causing human infections. The study provides a demonstration of the advantages and potential of the upconverting phosphor-based reporter technology in multianalyte assays and anti-Stokes photoluminescence detection with an anti-Stokes photoluminescence imaging device.

Photonen aufkonvertierende Nanopartikel zur optischen Codierung und zum Multiplexing von Zellen, Biomolekülen und Mikrosphären

“…Die Gruppe von Soukka [206][207][208] [209] 9. Durch Mikrosphären codierte heterogene Assays Die Mikrotiterplatte als Festphase für Immunassays kann durch Mikrosphären ersetzt werden, und suspendierte Mikrosphären kçnnen durch Durchflusszytometrie sortiert oder in Femtoliterarrays eingeordnet werden, wie in der Einleitung beschrieben.…”

Upconverting Phosphor Labels for Bioanalytical Assays

“…Ylihärsilä et al [136] demonstrated genotyping of ten common adenovirus genotypes with an array-in-well platform and validated the assay with clinical samples. Eleven selective probe spots (diameter 375 μm) in three replicas and positive control oligonucleotide spots were printed on the bottom of standard microtiter wells.…”

“…OraSure Technologies, Inc. (former STC Technologies; Pennsylvania, USA) with its research partners SRI International (California, USA) and Leiden University Medical Center (the Fig. 6 Array-in-well platform for qualitative genotyping based on hybridization patterns [136]. The DNA probes were spotted in three replicas together with control spots in the corners.…”

“…Commonly, the research groups have utilized standard spectrofluorometers equipped with an appropriate external excitation source (omitted from Table 5). Similarly, FluoroCount™ [139,140] and Plate Chameleon™ fluorometers [136,203] have been modified by changing certain components resulting in dedicated instrumentation for UCP detection. The cuvette-based multiparameter measurement apparatus developed at Chinese Academy of Sciences (Shanghai, China) has a special feature as it is has been designed to quantify the UCP concentration based on scattering in addition to luminescence measurement [204].…”

“…The PlateChameleon Imager [136] has a significantly more powerful LD compared to the other instruments in Table 5. The high power is required to compensate the low power density due to a relatively large illumination area covering a standard microwell bottom.…”