Apparatus and method for overcoming minimal mass sensitivity limitations in a shear horizontal surface acoustic wave biosensor
Inventors
Shachar, Josh • Kornberg, Roger
Assignees
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Publication Number
US-11293919-B2
Publication Date
2022-04-05
Expiration Date
Abstract
The invention includes a method of assaying an analyte in a sample in a portable, handheld microfluidic reader. The method includes the steps of: inserting the sample in the reader; capturing the analyte with a first antibody having a DNA tag attached thereto; capturing the analyte in the sample with a second antibody attached to a surface or having a magnetic nanoparticle (MNP) attached thereto; where a sandwich including the magnetic nanoparticle, first and second antibodies, the analyte and the DNA tag is formed; replicating the DNA tag using isothermal amplification to a predetermined amount of DNA tags detectable by a detector sufficient to overcome the minimal mass sensitivity limitations of the detector; and measuring the amount of replicated DNA tags using the detector. The invention also includes an apparatus or handheld portable field microfluidic reader in which the method is performed.
Core Innovation
The invention relates to a method of assaying an analyte in a sample in a portable, handheld microfluidic reader having a detector with a minimal mass sensitivity limitation. A rotatable microfluidic disc is used with a sample inlet, a mixing chamber, an amplification chamber, and a detector selectively communicated to the amplification chamber. The mixing chamber is provided with a first antibody for capturing the analyte having a DNA tag attached thereto.
An amplification chamber defined in the disc is selectively communicated to the mixing chamber and includes a second antibody for capturing the analyte attached to a surface or having a magnetic nanoparticle (MNP) or nanoparticle (NP) attached thereto. In the amplification chamber, a sandwich is formed including the surface or MNP or NP, first and second antibodies, the analyte, and the DNA tag. The DNA tag is replicated using isothermal amplification to produce a predetermined amount of DNA tags.
The predetermined amount of DNA tags is produced to be sufficient to overcome the minimal mass sensitivity limitation of the detector by providing an amount reliably detectable by the detector. The amplified DNA tags are then measured by the detector that is selectively communicated to the amplification chamber. The reader is also directed to using the rotatable disc workflow including formation of the sandwich, replication of the DNA tag, and measuring the amount of replicated DNA tags.
Claims Coverage
The document contains two independent claims describing a portable handheld microfluidic reader with a rotatable microfluidic disc, antibody-based DNA tagging, isothermal amplification of the DNA tag to a predetermined detectable amount, and detector measurement configured to address detector minimal mass sensitivity limitation. The inventive features center on sandwich formation with a DNA tag and a second antibody on a surface or MNP/NP, isothermal replication of the DNA tag to a predetermined amount, and detector measurement selectively communicated with the amplification chamber.
Portable handheld microfluidic reader with rotatable disc workflow addressing minimal mass sensitivity
A method of assaying an analyte in a sample in a portable, handheld microfluidic reader having a detector with a minimal mass sensitivity limitation, the reader having a rotatable microfluidic disc with a sample inlet, a mixing chamber selectively communicated to the sample inlet, an amplification chamber selectively communicated to the mixing chamber, and a detector selectively communicated to the amplification chamber for measuring the amount of replicated DNA tags.
Sandwich formation using a first antibody DNA tag and a second antibody attached to a surface or MNP/NP
Capturing the analyte with a first antibody having a DNA tag attached thereto and with the second antibody having an attached magnetic nanoparticle (MNP) or nanoparticle (NP), where a sandwich is formed including the first and second antibodies, the analyte, the MNP, the NP, and the DNA tag; the amplification chamber provides the sandwich including the surface or MNP or NP, first and second antibodies, the analyte and the DNA tag.
Isothermal amplification to a predetermined amount sufficient to overcome detector minimal mass sensitivity
Replicating the DNA tag using isothermal amplification to produce a predetermined amount of DNA tags sufficient to overcome the minimal mass sensitivity limitation of the detector by providing an amount reliably detectable by the detector.
Measuring the replicated DNA tags using a detector selectively communicated to the amplification chamber
Measuring the amount of replicated DNA tags using the detector that is selectively communicated to the amplification chamber.
Portable, handheld microfluidic reader apparatus with sample inlet, mixing chamber, amplification chamber, and detector
A portable, handheld microfluidic reader for assaying an analyte in a sample having a rotatable microfluidic disc comprising a sample inlet, a mixing chamber with a first antibody for capturing the analyte having a DNA tag attached thereto, an amplification chamber with a second antibody for capturing the analyte attached to a surface or having a magnetic nanoparticle (MNP) or nanoparticle (NP) attached thereto where a sandwich including the surface or MNP or NP, first and second antibodies, the analyte and the DNA tag are formed, and a detector selectively communicated to the amplification chamber and provided in the disc for measuring the amount of replicated DNA tags.
Isothermal amplification of DNA tags to produce a predetermined amount measurable by the detector
Replicating the DNA tag using isothermal amplification to produce a predetermined amount of DNA tags; with the detector for measuring the amount of replicated DNA tags.
Across the independent claims, the inventive concept is that an antibody immuno-sandwich incorporating a DNA tag is formed in an amplification chamber of a portable handheld microfluidic reader on a rotatable disc, the DNA tag is replicated by isothermal amplification to a predetermined amount sufficient to overcome a detector minimal mass sensitivity limitation, and the resulting replicated DNA tags are measured by a detector selectively communicated with the amplification chamber.
Stated Advantages
Overcomes the detector minimal mass sensitivity limitation by producing a predetermined amount of DNA tags that is reliably detectable by the detector.
Documented Applications
Assaying analyte in a sample using a portable, handheld microfluidic reader, including analyte detection based on replicated DNA tags.
Viral particle detection including HIV p24/p36, HIV2 p41, and Ebola Zaire virus p41.
Detection of troponin (cTnI/T).
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