PATENT CLAIM ANALYSIS

Application Number: 16313906
Application Type: Utility
Filing Date: 2018-12
Publication Date: 2019-10
Patent Classification: ["506", "009000"]

Abstract:
A method for detecting trace fungi using single-cell sequencing and a fungi detection kit prepared by the method are disclosed. The method includes the steps of obtaining trace fungal cells, extracting fungal protoplasm by breaking the cell walls of fungi, extracting gDNA from trace fungal protoplasm and amplifying the gDNA, constructing trace gDNA library, genome sequencing, bioinformatics analysis and comparison, and determining the species of detected fungi, etc. The method realizes the high-efficiency detection of trace fungi, and can be directly applied to the isolation, detection, identification of trace and difficult-to-identify fungal samples or mixed samples, and in-depth study of genetic information. The fungi detection method and the kit are applicable to industrial production, environmental monitoring, air testing, soil testing, water quality testing, food testing, drug testing, cosmetics testing, health care products testing and medical testing, and other fields.

Claim (Index 10):
The method for detecting the trace fungi using the single-cell sequencing according to  claim 1 , wherein the step (2) is as follows:\n (2) wall-breaking of the fungi cell walls: extraction of the fungal protoplasm by the chemical method combined with the mixed enzymatic method:\n 1) adding the 0.8 M D-sorbitol solution into the centrifuge tube containing the target cells to immerse the target cells at 4\u00b0 C. for 2 h; \n 2) preparation of the composite pretreatment agent: mixing the 50 mM Tris, the 5 mM EDTA and the 5% \u03b2-mercaptoethanol evenly; \n 3) preparation of the mixed enzyme treatment agent: mixing the 5 mg/mL lywallzyme, the 4 mg/mL of lysozyme, and the 3 mg/mL of cellulase; \n 4) adding the composite pretreatment agent into the centrifuge tube to treat the target cells at 35\u00b0 C. for 1 h; \n 5) subjecting the centrifuge tube to the centrifugation to collect the target cells to the bottom of the centrifuge tube, then discarding the composite pretreatment agent; \n 6) adding the sterile water to wash the target cells twice, centrifuging, and discarding the liquid; \n 7) adding the mixed enzyme treatment agent into the centrifuge tube to treat the target cells at 37\u00b0 C. for 6 h; \n 8) subjecting the centrifuge tube to the centrifugation to collect the target cells to the bottom of the centrifuge tube, then discarding the liquid; and \n 9) adding the PBS into the centrifuge tube to wash the target cells once, centrifuging, discarding the liquid, and retaining 10 \u03bcl of the cell suspension.

Metadata:
- Claim Count in Document: 1.0
- Percentile: 98.0
- Lexical Diversity: 2.01235
- Patent Class: 506.0
- Transitional Phrase Type: none
- Component Type: 0
- Foreign Priority: True
- Related Applications: ['14117035', '11212812', '14648161', '15612091', '13833872']

Analysis Scores:
- 35 USC 101 Eligibility (BERT): 0.5314266753350124
- 35 USC 102 Novelty (BERT): 0.500639608008099
- Combined Prediction Score: 0.5283479686023211
- Mean Citation Score: 175.581828
- Max Citation Score: 194.80453
- Similarity Product: 129.23011939956962

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

Dataset: test