PATENT CLAIM ANALYSIS

Application Number: 16158181
Application Type: Utility
Filing Date: 2018-10
Publication Date: 2019-03
Patent Classification: ["435", "006100"]

Abstract:
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.

Claim (Index 1):
A method for identifying pathogens in a plant sample comprising the steps of:\n a) harvesting pathogens from a plant tissue sample; b) isolating total nucleic acids comprising DNA; c) performing a first amplification in tandem in a single assay in the presence of plant DNA and non-DNA nucleic acids using at least one first primer pair selective for DNA isolated from the pathogens, to obtain one or more pathogen-specific first amplicons; d) performing a second amplification in tandem using the pathogen-specific first amplicons as a template and at least one first fluorescent labeled second primer pair having a sequence complementary to an internal flanking region in the first amplicons to obtain first fluorescent labeled second amplicons; e) hybridizing the second amplicons with nucleic acid probes specific for signature sequence determinants in the pathogen DNA, said nucleic acid probes immobilized at specific known positions on a 3-dimensional lattice microarray via a second fluorescent labeled bifunctional polymer linkers; f) washing the microarray at least once; g) imaging the microarray to detect a first fluorescent signal corresponding to the first fluorescent labeled second amplicons and a second fluorescent signal corresponding to the nucleic acid probes immobilized at the specific known positions on the microarray via the second fluorescent labeled bifunctional polymer linkers; h) superimposing the first fluorescent signal with the second fluorescent signal to obtain a superimposed signal image; and i) comparing the sequence of the nucleic acid probe at one or more superimposed signal positions on the microarray with a database of signature sequence determinants for a plurality of pathogen DNA, thereby identifying the pathogens in the sample.

Metadata:
- Claim Count in Document: 15.0
- Percentile: 97.0
- Lexical Diversity: 1.71233
- Patent Class: 435.0
- Transitional Phrase Type: open
- Component Type: 1
- Foreign Priority: False
- Related Applications: ['16158276', '15916036', '12100519', '09865807', '14694008']

Analysis Scores:
- 35 USC 101 Eligibility (BERT): 0.6535220675419539
- 35 USC 102 Novelty (BERT): 0.6234628607581866
- Combined Prediction Score: 0.6505161468635772
- Mean Citation Score: 492.178766
- Max Citation Score: 703.1072
- Similarity Product: 495.7039330627442

Labels:
- Claim Label 101: 1
- Claim Label 102: 1
- Claim Label 103: 1
- Claim Label 112: 0
- Combined Label: 1
- Label 101 Adjusted: 1

Dataset: test