Opportunity
Short, single-stranded nucleic acids often serve as important factors for determining a disease or bioterrorism agent. Many platforms have been developed for detecting nucleic acid molecules with specific sequences, including polymerase chain reaction (PCR), bio-barcode-based detection and electrochemistry. PCR-based methods, in particular, have been largely used to detect nucleic acids in ultralow abundance. However, this approach requires labor-intensive procedures and cumbersome instrumentation. These limitations have created significant challenges in healthcare medication in developing countries and other resource-limited sites. Thus, effective detection and diagnosis, suitable for low-resource settings, is of particular importance.
In recent years, the development of visual detection methods based on gold nanoparticles (AuNPs), silver nanoparticles and graphene oxide has increased rapidly because of their simplicity and the visual readouts produced. Numerous methods based on AuNP aggregation have been developed to detect DNAs/RNAs, proteins and metal ions. However, the sensitivity is insufficient for detecting low-abundance target molecules. To improve sensitivity, enzymatic amplification and biobarcode amplification methods have been developed. However, each of the amplification methods has its own set of drawbacks.
The present invention provides novel and highly advantageous methods for detecting a target nucleic acid sequence even in low abundance, which are especially suitable for further diagnosis and prognosis of a disease or a health condition of a subject.
Technology
The present invention relates to methods for determining the presence or amount of a target nucleic acid sequence in a sample, in particular based on a magnetophoretic assay. The present invention also pertains to a kit comprising a magnetic particle and a polystyrene particle and is suitable for a magnetophoretic assay for determining the presence or amount of a target nucleic acid sequence.
A method for determining the presence or amount of a target nucleic acid sequence in a sample, uses magnetic particles loaded with a probe and polystyrene particles loaded with a probe, in particular a different probe, for directly or indirectly detecting the presence or amount of the target nucleic acid sequence which is in particular indicative of a disease. The methods of the present invention allow for determining target nucleic acid sequences even in low abundance and in complex samples with advantageous sensitivity and, thus, represent promising approaches which can in particular be used in the diagnosis of several diseases and health conditions. A kit comprising a loaded magnetic particle and a loaded polystyrene particle is provided and can be used for the methods of the present invention, in particular for diagnosis or prognosis of a disease in a subject such as a human.
Advantages
- For an easy, fast and reliable visual determination of a target nucleic acid sequence even in complex samples and even if it is present in low abundance in the sample.
- The micro-meter size of it further contributes to an easy and direct visual determination of the presence of the target nucleic acid sequence in a sample.
Applications
- For further diagnosis and prognosis of a disease or a health condition of a subject.
