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公开(公告)号：US20220333099A1
公开(公告)日：2022-10-20
申请号：US17855067
申请日：2022-06-30
IPC分类号： C12N15/10 , C12Q1/6837 , G01N33/53
摘要： Provided herein is a method for manufacturing a microarray system, for example, 3-dimensional lattice microarray system, for DNA sequence detection and analysis. A solid support, such as a plastic substrate, is contacted with a formulation containing a plurality of nucleic acid probes, a plurality of bifunctional polymer linkers, such as oligothymidine linkers, and a solvent mixture of water and a water-miscible liquid. The bifunctional polymer linkers are attached to the solid support and the water is evaporated. Then the nucleic acid probes are attached to the bifunctional polymer linker.
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公开(公告)号：US20220267867A1
公开(公告)日：2022-08-25
申请号：US17529666
申请日：2021-11-18
申请人： Frederick Henry Eggers , Benjamin Alan Katchman , Fushi Wen , Candy Mavis Rivas , Cory Scott Newland , Michael Edward Hogan
发明人： Frederick Henry Eggers , Benjamin Alan Katchman , Fushi Wen , Candy Mavis Rivas , Cory Scott Newland , Michael Edward Hogan
摘要： Provided herein is a method for detecting the presence of clade variants in the COVID-19 virus in a human sample and/or an environmental sample. Samples are processed to obtain total RNA. The RNA is used as a template in a combined reverse transcription and amplification reaction to obtain fluorescent COVID-19 virus amplicons. These amplicons are hybridized on a microarray with nucleic acid probes having sequences that discriminate among the various clade variants. The microarray is imaged to detect the clade variant and each clade variant is distinguished from others by generating an intensity distribution profile from the image, which is unique to each of the clade variants.
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公开(公告)号：US20220195539A1
公开(公告)日：2022-06-23
申请号：US16950171
申请日：2020-11-17
IPC分类号： C12Q1/70
摘要： Provided herein is a method for detecting the presence of a COVID-19 virus in a human sample or an environmental sample having one or more viral and bacterial pathogens. Samples processed to obtain total nucleic acids. The nucleic acids are used as a template in a reverse transcription-amplification reaction to obtain cDNA, which is used in a PCR amplification reaction to obtain fluorescent COVID-19 virus specific amplicons. These amplicons are detected by microarray hybridization near the lowest limit of detection. Also provided is a method for detecting in addition to the COVID-19 virus, the presence of respiratory disease-causing pathogens including viruses, bacteria and fungus in a single assay using the above method.
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公开(公告)号：US20200283845A1
公开(公告)日：2020-09-10
申请号：US16819564
申请日：2020-03-16
IPC分类号： C12Q1/6874
摘要： Provided is a method for a two-step multiplex DNA amplification reaction which allows bacterial or fungal DNA analysis without first extracting DNA from the sample, nor without need to enrich microbes by laboratory culture prior to analysis. Without additional DNA purification or analysis, the PCR amplified DNA is administered directly to a microarray designed to interrogatea large panel of meaningful bacteria or fungi as a single multiplex test. Microarray analysis is then performed at ambient temperature, thus enabling substantial simplification of the testing process. It is contemplated that analysis may be conducted on unprocessed and processed leaf wash and similar surface sampling of plant material, cannabis, vegetables, fruit, nuts, spices, grains, other agriculture samples, food samples, or water samples, so as to detect bacterial, yeast, mold or viral, plant or human pathogen contamination.
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公开(公告)号：US20180251822A1
公开(公告)日：2018-09-06
申请号：US15916036
申请日：2018-03-08
IPC分类号： C12Q1/6818
CPC分类号： C12Q1/6818 , C12Q1/6837 , C12Q1/689 , C12Q1/6895 , C12Q2531/113 , C12Q2549/119 , C12Q2563/107 , C12Q2565/549
摘要： Provided herein is a dual amplification method for detecting plant pathogens by analysis of pathogen DNA in an unpurified nucleic acid sample from the plant. Pathogen-specific primers are used to generate a first set of amplicons that are further amplified in a second amplification step using fluorescent tagged pathogen-specific primers. Fluorescent amplicons thus generated are hybridized with pathogen-specific nucleic acid probes that are immobilized on a solid support using bifunctional polymer linkers. The hybridized microarray is imaged to obtain fluorescent images of the amplicons and the nucleic acid probes, which are superimposed to detect the pathogen present in the plant. Also described herein is a method to simultaneously detect both plant DNA and pathogen DNA in a single assay.
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公开(公告)号：US20220364157A1
公开(公告)日：2022-11-17
申请号：US16950210
申请日：2020-11-17
IPC分类号： C12Q1/6816 , C12Q1/686 , C12Q1/6876
摘要： Provided herein is a method for detecting the presence of a COVID-19 virus in a human sample or an environmental sample having one or more viruses and bacterial pathogens. Samples are processed to obtain total nucleic acids. A combined reverse transcription and asymmetric PCR amplification reaction is performed to obtain fluorescent labeled COVID-19 virus specific amplicons. The amplicons are detected by microarray hybridization near the lowest limit of detection. Also provided is a method for detecting concurrently with COVID-19 virus, the presence of respiratory disease-causing pathogens including viruses, bacteria and fungus in a single assay using the above method.
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公开(公告)号：US10513745B2
公开(公告)日：2019-12-24
申请号：US16158276
申请日：2018-10-11
IPC分类号： C12Q1/6895 , C12Q1/689 , C12Q1/686 , C12Q1/6837 , C12Q1/6893 , C12Q1/6874 , C12Q1/6844 , C12Q1/682 , C12Q1/6853 , G16B30/00 , G16B25/10 , G16B20/00
摘要： Provided herein is an internal standard method for determining copy number of a pathogen DNA in an unpurified nucleic acid sample by using a known copy number of synthetic DNA that shares a consensus region sequence with the pathogen. The sample is subject to two amplification steps using locus-specific primers and fluorescent primers respectively to obtain fluorescent amplicons for the pathogen and synthetic DNA. These are hybridized with immobilized pathogen-specific and synthetic DNA-specific nucleic acid probes and imaged to obtain fluorescent signals for pathogen-specific and synthetic DNA-specific amplicons. Signal intensities are correlated with the known copy number of synthetic DNA to determine copy number of pathogen DNA in the plant. Also described herein is a method to simultaneously quantitate using the above method, copy numbers of both pathogen and plant DNA in a sample.
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公开(公告)号：US20190032046A1
公开(公告)日：2019-01-31
申请号：US16157404
申请日：2018-10-11
IPC分类号： C12N15/10
摘要： Provided herein is a method for manufacturing a 3-dimensional lattice microarray system for DNA sequence detection and analysis. A solid support is contacted with a formulation containing a plurality of nucleic acid probes, a plurality of bifunctional polymer liners and a solvent mixture of water and a water-miscible liquid. In a first attachment reaction the bifunctional polymer linkers are attached to the solid support and the water is evaporated. In a second attachment reaction the nucleic acid probes are attached to the bifunctional polymer linker.
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公开(公告)号：US12098434B2
公开(公告)日：2024-09-24
申请号：US16950171
申请日：2020-11-17
IPC分类号： C12Q1/70
CPC分类号： C12Q1/701 , C12Q2600/112
摘要： Provided herein is a method for detecting the presence of a COVID-19 virus in a human sample or an environmental sample having one or more viral and bacterial pathogens. Samples processed to obtain total nucleic acids. The nucleic acids are used as a template in a reverse transcription-amplification reaction to obtain cDNA, which is used in a PCR amplification reaction to obtain fluorescent COVID-19 virus specific amplicons. These amplicons are detected by microarray hybridization near the lowest limit of detection. Also provided is a method for detecting in addition to the COVID-19 virus, the presence of respiratory disease-causing pathogens including viruses, bacteria and fungus in a single assay using the above method.
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公开(公告)号：US11781133B2
公开(公告)日：2023-10-10
申请号：US17855067
申请日：2022-06-30
IPC分类号： C12N15/10 , C12Q1/6837 , G01N33/53
CPC分类号： C12N15/1086 , C12Q1/6837 , G01N33/5308
摘要： Provided herein is a method for manufacturing a microarray system, for example, 3-dimensional lattice microarray system, for DNA sequence detection and analysis. A solid support, such as a plastic substrate, is contacted with a formulation containing a plurality of nucleic acid probes, a plurality of bifunctional polymer linkers, such as oligothymidine linkers, and a solvent mixture of water and a water-miscible liquid. The bifunctional polymer linkers are attached to the solid support and the water is evaporated. Then the nucleic acid probes are attached to the bifunctional polymer linker.
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