For example, the CDC has been using advanced molecular detection technology, such as whole-genome sequencing as means of surveying the genetic differences between isolates in various pathogenic organisms.
Samples were then incubated with goat anti-rabbit Alexa Fluor 488 (Molecular Probes, Invitrogen Detection Technologies, Eugene, OR, USA; 1 200), goat anti-mouse Alexa Fluor 594 (Molecular Probes, CA, USA; 1 200), and DAPI (Molecular Probes 1 10 000) for 1 h at room temperature.
The mouse IgG secondary antibody conjugated with Oregon green was used to detect mouse monoclonal antibody against p53 (DO-1) and the rat IgG secondary antibody conjugated with Texas red was used to detect rat monoclonal antibody against Hsp90α (Molecular Probes, Invitrogen Detection Technologies).
The DNA obtained was quantified using a Qubit fluorometer apparatus (Molecular Probes, Invitrogen Detection Technologies, Oregon, USA).
To investigate apoptosis annexinV Alexa Fluor®647 in combination with PI were added (Vybrant® Apoptosis Assay Kit, Molecular ProbesTM Invitrogen detection technologies).
Fixed nuclei were mounted on SuperFrost-plus slides (Fisher Scientific, Pittsburgh, PA) with Prolong Gold antifade reagent with DAPI (Molecular Probes, Invitrogen Detection Technologies, Carlsbad, CA).
Cell nuclei were then stained with 0.5 μg/μL propidium iodide [P21493 (Molecular Probes, Invitrogen Detection Technologies)] for 5 min. The fluorescence-labeled cells on microscope slides were mounted with ProLong Gold Antifade reagent [P36930 (Life Technologies Corp., Invitrogen)] and sealed with nail polish.
SERS was a very sensitive technology for molecular detection, and it is particularly important to select a suitable SERS substrate.
The range of nucleic acid-based technologies for the molecular detection of pathogens has grown rapidly in recent years.
