PATENT CLAIM ANALYSIS

Application Number: 15877007
Application Type: Utility
Filing Date: 2018-01
Publication Date: 2018-08
Patent Classification: ["435", "006120"]

Abstract:
This invention discloses multi-part primers for primer-dependent nucleic amplification methods. Also disclosed are primer-dependent nucleic acid amplification reactions, particularly DNA amplification reactions, reaction mixtures and reagent kits for such reactions. This invention relates to primer-dependent nucleic acid amplification reactions, particularly DNA amplification reactions such as PCR, and primers, reaction mixtures and reagent kits for such reactions and assays employing same.

Claim (Index 24):
A kit of reagents for performing a homogeneous primer-dependent amplification and homogeneous detection method that includes multiple cycles of primer-annealing and extension, said method being capable of detecting as few as ten copies of at least one rare mutant DNA target sequence in a mixture containing, for each mutant target sequence, 100,000 copies of a closely related wild-type DNA target sequence, said kit comprising a DNA polymerase; amplification buffer; dNTPs; for each mutant target sequence a primer pair that includes a reverse primer and a\n multi-part primer comprising, in the 5\u2032 to 3\u2032 direction the following three contiguous DNA sequences:\n an anchor sequence that hybridizes with the mutant target sequence and with its closely related wild-type target sequence during primer annealing, \n a bridge sequence at least six nucleotides long that does not hybridize to either the mutant target sequence or its closely related wild-type target sequence during primer annealing, and \n a foot sequence that is at least five nucleotides long, perfectly complementary to the mutant sequence and mismatched to its wild-type sequence by one or two nucleotides; and \n a fluorescent detection reagent for detecting the amplified product of the at least one multi-part primer; wherein:\n (i) if the anchor sequence and the foot sequence of the primer are hybridized either to the mutant target sequence or to its closely related wild-type target sequence to form a hybrid, there is in the target sequence an intervening sequence at least six nucleotides long that does not hybridize to the primer's bridge sequence during primer-annealing, and the bridge sequence and the intervening sequence, together create a bubble in the hybrid having a circumference of 16-52 nucleotides, \n (ii) the circumference of the bubble and the length of the foot sequence in combination result in a weak foot/mutant-target-sequence hybrid that makes copying the intended target sequence unlikely as evidenced by a delay of at least two cycles in the threshold cycle (C T ) of amplification of said at least one mutant target sequence using said multi-part primer as compared to using a conventional primer, \n (iii) the bridge sequence and the foot sequence do not together prime non-target sequences, and \n (iv) the probability that during a cycling of the primer-dependent amplification reaction a multi-part primer/wild-type-target-sequence hybrid will be extended is at least 10,000 times lower than the probability that during said cycling a multi-part primer/mutant-target-sequence hybrid will be extended, as evidenced by a \u0394C T  of at least 13.3 cycles.

Metadata:
- Claim Count in Document: 65.0
- Percentile: 86.0
- Lexical Diversity: 2.125
- Patent Class: 435.0
- Transitional Phrase Type: open
- Component Type: 1
- Foreign Priority: False
- Related Applications: ['14766139', '15393103', '10632393', '10328150', '12185699']

Analysis Scores:
- 35 USC 101 Eligibility (BERT): 0.6279892732512043
- 35 USC 102 Novelty (BERT): 0.6355515396589042
- Combined Prediction Score: 0.6287454998919744
- Mean Citation Score: 385.617464
- Max Citation Score: 679.42566
- Similarity Product: 548.5512611408615

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

Dataset: test