Abstract
The spin-column system for the isolation of nucleic acids (NAs) from multiple samples presents the inconvenience of repeated experimentation, time-consumption, and the risk of contamination in the process of the spin-column exchange. Herein, we propose a convenient and universal assay that can be used to diagnose multiple pathogens using a multi-sample preparation assay. The multi-sample preparation assay combines a 96-well filter/membrane plate, a bio-micromaterial lattice-like micro amine-functional diatomaceous earth (D-APDMS), and homobifunctional imidoesters (HI) for the processing of pathogen enrichment and extraction for multiple samples simultaneously. The purity and quantity of the extracted NAs from pathogens (E. coli and Brucella) using the proposed assay is superior to that of the commercialized spin-column kit. The assay also does not require the replacement of several collection tubes during the reaction processing. For the multi-sample testing, we used as many as six samples simultaneously with the proposed assay. This assay can simultaneously separate up to 96 NAs from one plate, and the use of multichannel pipettes allows faster and simpler experimentation. Therefore, we believe it is a convenient and easy process, and can be easily integrated with other detection methods for clinical diagnostics.
Highlights
The entire world pays tribute to all medical staff working on the front lines of infectious disease diagnosis, especially during rapid virus and pathogen infections [1,2]
We developed a convenient multi-sample preparation assay by uniting diatomaceous earth (DE) having micro-grid structure and 96-well filter/membrane plates, which could enrich the target samples for accurate diagnosis
The schematic diagram of pathogen enrichment with the micro lattice-like D-APDMS and homobifunctional imidoesters (HI) reagent, in the tube is shown in (Figure 1i). The imidoesters at both ends of the DMP help to form an amide bond with the amine group exposed on the surface of the D-APDMS and support to combine pathogens or nucleic acids (NAs)
Summary
The entire world pays tribute to all medical staff working on the front lines of infectious disease diagnosis, especially during rapid virus and pathogen infections [1,2]. Rapid diagnostic techniques, combined with automation and/or a self-examination system, are increasingly expected to save labor as well as time and provide better protection of the medical inspectors [5]. The spin-column based sample preparation technology is popular for nucleic acid (NA) extraction. It is an expensive kit and experienced technicians are required during the experiment. For NA extraction from multi-samples, the spin-column based sample preparation is unsuitable and labor intensive [7]. To address this issue, we used 96-well filter/membrane plates to extract NAs from multiple samples simultaneously. Multichannel pipettes can be more convenient for conducting experiments on various samples simultaneously
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
