Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Election/Restrictions
Applicant’s election without traverse of species in the reply filed on 6/9/2025 is acknowledged. The Applicant failed to elect one species from the group of species in group (i) and one species from group (ii), as required in the Office action dated 4/9/2025 (page 3, last line). The Applicant elects all 4 species comprising SEQ ID NOs: 1-4 among the 8 species of group (i).
However, the Examiner rejoins all the species and withdraws the restriction requirement.
Claim Status
Claims 1-17 are pending and all claims are being examined.
Claim Rejections - 35 USC § 112(b)
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claim 1-17 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claims 1-2, 5-6, and 16-17 recites, “a nucleotide sequence… represented by SEQ ID NO:…”.
It is not clear if Nanovirus DNA-C “represented by” SEQ ID NO: 1 encompasses nucleic acids that comprise the full-length sequence of SEQ ID NO: 1 or any portion of SEQ ID NO: 1. The same lack of clarity is present for Nanovirus DNA-M, Nanovirus DNA-N, Nanovirus DNA-U1, Nanovirus DNA-R, and Nanovirus DNA-S components and the corresponding SEQ ID NOs.
Claims 1 recites, “… a nucleotide sequence of Nanovirus DNA-C represented by SEQ ID NO: 1; a nucleotide sequence of Nanovirus DNA-M represented by SEQ ID NO: 2; …”.
Similarly, independent claim 2 recites, “… a nucleotide sequence of Nanovirus DNA-C represented by SEQ ID NO: 5; a nucleotide sequence of Nanovirus DNA-M represented by SEQ ID NO: 6; …”.
Independent claim 7 recites, “… (a) an agent for detecting a Nanovirus comprising Nanovirus DNA-C including the nucleotide sequence of SEQ ID NO: 1 or a fragment thereof, Nanovirus DNA-M including the nucleotide sequence of SEQ ID NO: 2 or a fragment thereof;
(b) an agent for detecting a Nanovirus comprising Nanovirus DNA-C including the nucleotide sequence of SEQ ID NO: 5 or a fragment thereof, Nanovirus DNA-M including the nucleotide sequence of SEQ ID NO: 6 or a fragment thereof, …”
SEQ ID NO: 1 comprises 100% sequence identity to SEQ ID NO: 2, as shown below.
Title: US-18-523-237-1
Perfect score: 988
Sequence: 1 ctggggcggggcttagtatt..........catatgatcgaggtgctgag 988
Searched: 1 seqs, 988 residues
Database : US-18-523-237-2.seq:*
RESULT 1
US-18-523-237-2
Query Match 100.0%; Score 988; DB 1; Length 988;
Best Local Similarity 100.0%;
Matches 988; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 CTGGGGCGGGGCTTAGTATTACCCCCGCCCCAGGCTCAGCGGAGTCATCACGTGAGACGC 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 CTGGGGCGGGGCTTAGTATTACCCCCGCCCCAGGCTCAGCGGAGTCATCACGTGAGACGC 60
Qy 61 ACATGACTATTAAATAGAAGACATAATACCAAGTGGTATATGGTCCCCACTAATATATTT 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 ACATGACTATTAAATAGAAGACATAATACCAAGTGGTATATGGTCCCCACTAATATATTT 120
Qy 121 GATTTAAATGCACGGAATATATGCTTTGCTTCGTCTCGAAGCAAAGTAAGGAATCACTGG 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 GATTTAAATGCACGGAATATATGCTTTGCTTCGTCTCGAAGCAAAGTAAGGAATCACTGG 180
Qy 181 GACCGATATTAGCTGTAACCCATGTGCCTTCCTTTATTTGTATGGTGGACATTAATTTGC 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 GACCGATATTAGCTGTAACCCATGTGCCTTCCTTTATTTGTATGGTGGACATTAATTTGC 240
Qy 241 TTTAAGATAGCGTACATTGGAAGCATTGTTATTGCTTATAAAACCTGAGCCGCTATTGTC 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 TTTAAGATAGCGTACATTGGAAGCATTGTTATTGCTTATAAAACCTGAGCCGCTATTGTC 300
Qy 301 ATTTTTGCTGTTCCGTCGTGTTTCTATTGTCCATTGATATTTCATGGCGGATCCTGGATA 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 ATTTTTGCTGTTCCGTCGTGTTTCTATTGTCCATTGATATTTCATGGCGGATCCTGGATA 360
Qy 361 TTATCAAGGTTATCAAGACGACGGCGATATCGATGCCCAGAAGAGACATCAAGCTCTATA 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 TTATCAAGGTTATCAAGACGACGGCGATATCGATGCCCAGAAGAGACATCAAGCTCTATA 420
Qy 421 TTTGATCGGCATAATTATACTCATCATGGTATGTATTATTATTCTTTGGGTTTGTATTAT 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 TTTGATCGGCATAATTATACTCATCATGGTATGTATTATTATTCTTTGGGTTTGTATTAT 480
Qy 481 GTTGGCGTGTTATATTCCTGGGTTTTTGAAGAAGACAATGGAAACGTGGTTAAGTTCATC 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 GTTGGCGTGTTATATTCCTGGGTTTTTGAAGAAGACAATGGAAACGTGGTTAAGTTCATC 540
Qy 541 ATCTATGATGAAGAGGCGTCTTGCTGCGACTATAACACGCACACCCTTCGAGGCAACAGG 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 ATCTATGATGAAGAGGCGTCTTGCTGCGACTATAACACGCACACCCTTCGAGGCAACAGG 600
Qy 601 ACCTGAGAGAGAACTAAATTGGGATGCTCGCAGACAAGCAAACCCTGCACCAAGTCAGCC 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 ACCTGAGAGAGAACTAAATTGGGATGCTCGCAGACAAGCAAACCCTGCACCAAGTCAGCC 660
Qy 661 TTCCAATGGAGGAGTATTTTAGAGAATGATTATAGATTGTAATTATATATTCATATCATC 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 TTCCAATGGAGGAGTATTTTAGAGAATGATTATAGATTGTAATTATATATTCATATCATC 720
Qy 721 ATTCTATTGTAAGTGTCGCACCATCAATGAGAATTTACAGAAATTCGCTGTGTCGTAATT 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 ATTCTATTGTAAGTGTCGCACCATCAATGAGAATTTACAGAAATTCGCTGTGTCGTAATT 780
Qy 781 GTGTTTCGTATGAGTTGCAGACTTCTGTATATGTTTCAAGGAAGAAGAAAACTATTCACT 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 GTGTTTCGTATGAGTTGCAGACTTCTGTATATGTTTCAAGGAAGAAGAAAACTATTCACT 840
Qy 841 ATGCCACGTGTTCACATAGAAATGGTTTATATTATTTTTTATTACTCCGCGTAGCGGTAT 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 ATGCCACGTGTTCACATAGAAATGGTTTATATTATTTTTTATTACTCCGCGTAGCGGTAT 900
Qy 901 GTTTGGGCTTTTAAGCCCAATTGGGGTTAGGCCCAATTCAGTAGCCAATGAGGAAGGCTT 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 GTTTGGGCTTTTAAGCCCAATTGGGGTTAGGCCCAATTCAGTAGCCAATGAGGAAGGCTT 960
Qy 961 AATGACCTCATATGATCGAGGTGCTGAG 988
||||||||||||||||||||||||||||
Db 961 AATGACCTCATATGATCGAGGTGCTGAG 988
Similarly, SEQ ID NO: 5 comprises 100% sequence identity to SEQ ID NO: 6, as shown below.
Title: US-18-523-237-5
Perfect score: 993
Sequence: 1 acccccgccccgggttcaga..........taggggcggggcttagtatt 993
Searched: 1 seqs, 993 residues
Database : US-18-523-237-6.seq:*
RESULT 1
US-18-523-237-6
Query Match 100.0%; Score 993; DB 1; Length 993;
Best Local Similarity 100.0%;
Matches 993; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 ACCCCCGCCCCGGGTTCAGAGTCACGTACGGAGTGACTAATGGCCATTGGATTATTATAC 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 ACCCCCGCCCCGGGTTCAGAGTCACGTACGGAGTGACTAATGGCCATTGGATTATTATAC 60
Qy 61 ACGTGGACGATCAGGATCTGTGATTCGTGAAGCGAATCTGACGGAAGATCGTCCGAAGCT 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 ACGTGGACGATCAGGATCTGTGATTCGTGAAGCGAATCTGACGGAAGATCGTCCGAAGCT 120
Qy 121 TCGTGGTAGGGCCCAAGTGTTGCTTTAGTTTGCTTTTTTAAAAAGTAAAGCTAGATGCTT 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 TCGTGGTAGGGCCCAAGTGTTGCTTTAGTTTGCTTTTTTAAAAAGTAAAGCTAGATGCTT 180
Qy 181 TTCTCTACTTTATTCCTTTGAAGTGGAATACAGCTGTCTTTGCTTCCTCCCGAAGCAAAG 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 TTCTCTACTTTATTCCTTTGAAGTGGAATACAGCTGTCTTTGCTTCCTCCCGAAGCAAAG 240
Qy 241 GATTCTTTATCTCTCTATAAAAAGCTGTGTTCTTCTTCGTTGTTCTTACAACGAAGATGG 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 GATTCTTTATCTCTCTATAAAAAGCTGTGTTCTTCTTCGTTGTTCTTACAACGAAGATGG 300
Qy 301 GTCTGAAATATTTTTCTCATCTTCCTGAAGAGTTGAGAGAGAAGATTGTTCGTGATCATC 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 GTCTGAAATATTTTTCTCATCTTCCTGAAGAGTTGAGAGAGAAGATTGTTCGTGATCATC 360
Qy 361 TACAACAAGAGCGAAAGAAAGAGTTTCTTGAGAATGCTATAGAAGATAGTTGCAGAAGGC 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 TACAACAAGAGCGAAAGAAAGAGTTTCTTGAGAATGCTATAGAAGATAGTTGCAGAAGGC 420
Qy 421 ATGTGTCGTTATTAAAGTCAGACCCATCACCTTCAGAGCTGTATTCTCTGAGTAAGTTTT 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 ATGTGTCGTTATTAAAGTCAGACCCATCACCTTCAGAGCTGTATTCTCTGAGTAAGTTTT 480
Qy 481 TAGATTCATTGGCTGATTATGTTGGTAAGCAGTTTAATACACGTTGTTTGATTAAATGGA 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 TAGATTCATTGGCTGATTATGTTGGTAAGCAGTTTAATACACGTTGTTTGATTAAATGGA 540
Qy 541 AGAAAGACGTTCCTGCTAATATCAAGTTTCAAGTTATGGAAGAACAACACCTGCGGTTAT 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 AGAAAGACGTTCCTGCTAATATCAAGTTTCAAGTTATGGAAGAACAACACCTGCGGTTAT 600
Qy 601 ATGGGTTTGTTGATATGGACGACCTTTTATGTCGTGAGTTATTGCCACCTGAAGAAGATG 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 ATGGGTTTGTTGATATGGACGACCTTTTATGTCGTGAGTTATTGCCACCTGAAGAAGATG 660
Qy 661 ATGATATTACATACGAAGATGGTATGATTGTCAACTGCAGTGAGTTGGATAAATTATTTG 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 ATGATATTACATACGAAGATGGTATGATTGTCAACTGCAGTGAGTTGGATAAATTATTTG 720
Qy 721 AGGCTCTAGGCATTAGAGTTGTTTATATTACTGTCAGTAAGAATTGTATTTGGACTCCTT 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 AGGCTCTAGGCATTAGAGTTGTTTATATTACTGTCAGTAAGAATTGTATTTGGACTCCTT 780
Qy 781 TGAACAAGGATATTGTAATTAAATGAATTGTGATAATAATAAATTATTACTAATGTTATC 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 TGAACAAGGATATTGTAATTAAATGAATTGTGATAATAATAAATTATTACTAATGTTATC 840
Qy 841 ATGAAAGATATATTTATTCTTTTAAATACTCTGCGAAGCTATATGTTAAGGCCCAATAGG 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 ATGAAAGATATATTTATTCTTTTAAATACTCTGCGAAGCTATATGTTAAGGCCCAATAGG 900
Qy 901 CCCAATTGCTCTTAGGCCCAATTAAATTCTCTTTGAAGTGAGTCAGCTGACATCAGCTGA 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 CCCAATTGCTCTTAGGCCCAATTAAATTCTCTTTGAAGTGAGTCAGCTGACATCAGCTGA 960
Qy 961 CTCCTTGATGACGTAGGGGCGGGGCTTAGTATT 993
|||||||||||||||||||||||||||||||||
Db 961 CTCCTTGATGACGTAGGGGCGGGGCTTAGTATT 993
Claims 1, 2, and 7, and all the claims dependent from these independent claims are rendered indefinite as it is not clear whether DNA-C sequence is represented by SEQ ID NO: 1 or SEQ ID NO: 2 of milk vetch dwarf virus (MDV). It is also not clear whether DNA-C sequence is represented by SEQ ID NO: 5 or SEQ ID NO: 6 of Faba bean necrotic yellow virus (FBNYV). The same lack of clarity is valid for DNA-M sequence for both the nanovirues- MDV and FBNYV.
In the spirit of compact prosecution, in the prior art rejection below, the limitation “a nucleotide sequence of … represented by SEQ ID NO” is interpreted that the “nucleotide sequence” consists of the entire sequence of the relevant SEQ ID NO.
Claim 9 depends from claim 7, and recites “the detecting agent comprises ….” This limitation lacks clear antecedent basis because there are multiple detecting agents recited in claim 7: various agents for detecting the different viruses in (a) and (b), and thus it is unclear which particular detecting agent is meant by “the detecting agent …” recited in claim 9.
Claim Rejections - 35 USC § 112(a)
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 1-6 and 14-17 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the enablement requirement. The claims contain subject matter which was not described in the specification in such a way as to enable one skilled in the art to which it pertains, or with which it is most nearly connected, to use the invention.
Independent claims 1-2 are drawn to a recombinant vector, comprising:
a nucleotide sequence of Nanovirus DNA-C represented by SEQ ID NO: 1 (for MDV) or SEQ ID NO: 5 (for FBNYV);
a nucleotide sequence of Nanovirus DNA-M represented by SEQ ID NO: 2 (for MDV) or SEQ ID NO: 6 (FBNYV);
a nucleotide sequence of Nanovirus DNA-N represented by SEQ ID NO: 3 (for MDV) or SEQ ID NO: 7 (FDNYV); and
a nucleotide sequence of Nanovirus DNA-U1 represented by SEQ ID NO: 4 (for MDV) or SEQ ID NO: 8 (for FBNYV).
The description teaches transforming a host crop plants with the recombinant vector “can effectively induce the infection of Nanovirus into crops even without any insect vectors” (abstract, line 4-5). The Applicant describes the invention as a development of an artificial infection system for nanovirus (spec, page 3, line 5-6). The Applicant describes using PCR and detecting the presence of the polynucleotide sequence of DNA-C, DNA-M, DNA-N, and DNA-U1 in MDV (page 27, line 19-22; Fig. 9a) and FBNYV (page 28, line 6-11; Fig. 9b) nanovirus infected plants which developed symptoms such as leaf necrosis, chlorosis, and stunted growth. However, Fig. 9 (a and b) does indicate presence of DNA-R, DNA-S, DNA-U2, and DNA-U4; besides DNA-C, DNA-M, DNA-N, and DNA-U1 as described.
Timchenko et al. (Infectivity of nanovirus DNAs: induction of disease by cloned genome components of Faba bean necrotic yellows virus, 2006, Journal of General Virology, 87:1735-1743) describes that five genome components, DNA-R, DNA-S, DNA-M, DNA-U1 and DNA-U2, were sufficient for inducing (infection and) disease symptoms in Fava bean (V. faba) upon agroinoculation (abstract). Grigoras et al. (Nanovirus DNA-N encodes a protein mandatory for aphid transmission, 2018, Virology 522:281–291) describes that DNA-R (master replication initiator protein; M-Rep) is required for replication initiation of all (viral) genomic DNAs (page 281, left column, para 1, line 11-13). DNA-R, DNA-S and DNA-M are “absolutely requited” (page 289, left column, para 3, line 8-10) and essential for infection and symptom development (page 522, right column, para 2). On the other hand, nanovirus DNA-N is mandatory for aphid transmission (abstract). A host plant transformed by a vector which does not include DNA-R and DNA-S would fail to effectively induce an infection of Nanovirus including MDV and FBNYV. However, the specific feature (DNA-R and DNA-S) is missing from the independent claims 1 and 2, and also missing from the dependent claims directly or indirectly depending from claim 1 or 2.
One skilled in the art would be able to make the claimed recombinant vectors but would not be able to use the invention commensurate with the scope of the invention as recited in the claims, in the spec, or both.
Claims 16-17 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for a recombinant vector comprising the nucleotide sequence of Nanovirus DNA-R represented by SEQ ID NO: 9 and Nanovirus DNA-S represented by SEQ ID NO: 10, does not reasonably provide enablement for a recombinant vector comprising the nucleotide sequence of Nanovirus DNA-R represented by SEQ ID NO: 9 or Nanovirus DNA-S represented by SEQ ID NO: 10. The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to use the invention commensurate in scope with these claims.
The description teaches transforming crop plants with the recombinant vector “can effectively induce the infection of Nanovirus into crops even without any insect vectors” (abstract, line 4-5). The Applicant describes development of an artificial infection system for nanovirus (spec, page 3, line 5-6). The Applicant describes detecting presence of DNA-C, DNA-M, DNA-N, and DNA-U1 in MDV (page 27, line 19-22; Fig. 9a) and FBNYV (page 28, line 6-11; Fig. 9b) infected plants, which developed symptoms such as leaf necrosis, chlorosis, and stunted growth.
However, Timchenko et al. describes that five genome components, DNA-R, DNA-S, DNA-M, DNA-U1 and DNA-U2, were sufficient for inducing (infection and) disease symptoms in Fava bean (V. faba) upon agroinoculation (abstract). Grigoras et al. describes that DNA-R (master replication initiator protein; M-Rep) is required for replication initiation of all (viral) genomic DNAs (page 281, left column, para 1, line 11-13). DNA-R, DNA-S and DNA-M are “absolutely requited” (page 289, left column, para 3, line 8-10) and essential for infection and symptom development (page 522, right column, para 2). On the other hand, nanovirus DNA-N is mandatory for aphid transmission for a Nanovirus (abstract). A crop plant transformed by a vector that includes DNA-C, DNA-M, DNA-N, and DNA-U1, along with either DNA-R or DNA-S would fail to induce an infection in the transformed plant. Both DNA-R and DNA-S are “absolutely required” to induce an infection in a host plant.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
Claims 7-13 are rejected under 35 U.S.C. 103 as being unpatentable over Lal et al. (Milk vetch dwarf virus infection in the Solanaceae and Caricaceae families in Southeast Asia, 2020, Plant Pathology, 69:1026–1033), in view of Zhang et al. (Complete genomic characterization of milk vetch dwarf virus isolates from cowpea and broad bean in Anhui province, China; 2017, Arch Virol., 162:2437–2440).
Independent claim 7 is drawn to a composition for diagnosis of Nanovirus infection, comprising:
(a) an agent for detecting a Nanovirus comprising Nanovirus DNA-C including the nucleotide sequence of SEQ ID NO: 1 or a fragment thereof, Nanovirus DNA-M including the nucleotide sequence of SEQ ID NO: 2 or a fragment thereof, Nanovirus DNA-N including the nucleotide sequence of SEQ ID NO: 3 or a fragment thereof, and Nanovirus DNA-U1 including the nucleotide sequence of SEQ ID NO: 4 or a fragment thereof,
(b) an agent for detecting a Nanovirus comprising Nanovirus DNA-C including the nucleotide sequence of SEQ ID NO: 5 or a fragment thereof, Nanovirus DNA-M including the nucleotide sequence of SEQ ID NO: 6 or a fragment thereof, Nanovirus DNA-N including the nucleotide sequence of SEQ ID NO: 7 or a fragment thereof, and Nanovirus DNA-U1 including the nucleotide sequence of SEQ ID NO: 8 or a fragment thereof; or
(c) a combination thereof.
Lal et al. also describes a nucleotide sequence of Nanovirus (MDV) DNA-M (GenBank accession No. MN795302) (page 1030, Table 1) having 100% sequence identity to SEQ ID NOs: 1-2, as shown below. (SEQ ID NO: 1 and SEQ ID NO: 2 share 100% identity to each other, as discussed above.)
RESULT 1
MN795302
LOCUS MN795302 988 bp DNA circular VRL 18-NOV-2020
DEFINITION Milk vetch dwarf virus isolate KS92 segment DNA M, complete sequence.
ACCESSION MN795302
VERSION MN795302.1
SOURCE Milk vetch dwarf virus
ORGANISM Milk vetch dwarf virus
Viruses; Monodnaviria; Shotokuvirae; Cressdnaviricota;
Arfiviricetes; Mulpavirales; Nanoviridae; Nanovirus.
REFERENCE 1 (bases 1 to 988)
AUTHORS Lal,A., Kil,E.-J., Thuy,V.T.B., Fadhila,C., Ho,P.T., Byun,H.-S.,
Dao,H.T., Kim,J.-K. and Lee,S.
TITLE Milk vetch dwarf virus infection in the Solanaceae and Caricaceae
families in Southeast Asia
JOURNAL Plant Pathol 69 (6), 1026-1033 (2020)
REMARK DOI: 10.1111/ppa.13196
REFERENCE 2 (bases 1 to 988)
AUTHORS Lal,A., Kil,E.-J., Vo,T.T.B., Fadhila,C., Ho,P.T. and Lee,S.
TITLE Direct Submission
JOURNAL Submitted (03-DEC-2019) Integrative Biotechnology, Sungkyunkwan
University, 2066, Seobu-ro, Jangan-gu, Suwon, Gyeonggi-do 16419,
South Korea
FEATURES Location/Qualifiers
source 1..988
/organism="Milk vetch dwarf virus"
/mol_type="genomic DNA"
/isolate="KS92"
/host="pepper"
/db_xref="taxon:67585"
/segment="DNA M"
/geo_loc_name="Viet Nam"
/collection_date="Aug-2018"
/collected_by="Sukchan Lee"
CDS 344..682
/codon_start=1
/product="movement protein"
/protein_id="QPB75528.1"
/translation="MADPGYYQGYQDDGDIDAQKRHQALYLIGIIILIMVCIIILWVC
IMLACYIPGFLKKTMETWLSSSSMMKRRLAATITRTPFEATGPERELNWDARRQANPA
PSQPSNGGVF
Query Match 100.0%; Score 988; Length 988;Best Local Similarity 100.0%;
Matches 988; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 CTGGGGCGGGGCTTAGTATTACCCCCGCCCCAGGCTCAGCGGAGTCATCACGTGAGACGC 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 CTGGGGCGGGGCTTAGTATTACCCCCGCCCCAGGCTCAGCGGAGTCATCACGTGAGACGC 60
Qy 61 ACATGACTATTAAATAGAAGACATAATACCAAGTGGTATATGGTCCCCACTAATATATTT 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 ACATGACTATTAAATAGAAGACATAATACCAAGTGGTATATGGTCCCCACTAATATATTT 120
Qy 121 GATTTAAATGCACGGAATATATGCTTTGCTTCGTCTCGAAGCAAAGTAAGGAATCACTGG 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 GATTTAAATGCACGGAATATATGCTTTGCTTCGTCTCGAAGCAAAGTAAGGAATCACTGG 180
Qy 181 GACCGATATTAGCTGTAACCCATGTGCCTTCCTTTATTTGTATGGTGGACATTAATTTGC 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 GACCGATATTAGCTGTAACCCATGTGCCTTCCTTTATTTGTATGGTGGACATTAATTTGC 240
Qy 241 TTTAAGATAGCGTACATTGGAAGCATTGTTATTGCTTATAAAACCTGAGCCGCTATTGTC 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 TTTAAGATAGCGTACATTGGAAGCATTGTTATTGCTTATAAAACCTGAGCCGCTATTGTC 300
Qy 301 ATTTTTGCTGTTCCGTCGTGTTTCTATTGTCCATTGATATTTCATGGCGGATCCTGGATA 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 ATTTTTGCTGTTCCGTCGTGTTTCTATTGTCCATTGATATTTCATGGCGGATCCTGGATA 360
Qy 361 TTATCAAGGTTATCAAGACGACGGCGATATCGATGCCCAGAAGAGACATCAAGCTCTATA 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 TTATCAAGGTTATCAAGACGACGGCGATATCGATGCCCAGAAGAGACATCAAGCTCTATA 420
Qy 421 TTTGATCGGCATAATTATACTCATCATGGTATGTATTATTATTCTTTGGGTTTGTATTAT 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 TTTGATCGGCATAATTATACTCATCATGGTATGTATTATTATTCTTTGGGTTTGTATTAT 480
Qy 481 GTTGGCGTGTTATATTCCTGGGTTTTTGAAGAAGACAATGGAAACGTGGTTAAGTTCATC 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 GTTGGCGTGTTATATTCCTGGGTTTTTGAAGAAGACAATGGAAACGTGGTTAAGTTCATC 540
Qy 541 ATCTATGATGAAGAGGCGTCTTGCTGCGACTATAACACGCACACCCTTCGAGGCAACAGG 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 ATCTATGATGAAGAGGCGTCTTGCTGCGACTATAACACGCACACCCTTCGAGGCAACAGG 600
Qy 601 ACCTGAGAGAGAACTAAATTGGGATGCTCGCAGACAAGCAAACCCTGCACCAAGTCAGCC 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 ACCTGAGAGAGAACTAAATTGGGATGCTCGCAGACAAGCAAACCCTGCACCAAGTCAGCC 660
Qy 661 TTCCAATGGAGGAGTATTTTAGAGAATGATTATAGATTGTAATTATATATTCATATCATC 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 TTCCAATGGAGGAGTATTTTAGAGAATGATTATAGATTGTAATTATATATTCATATCATC 720
Qy 721 ATTCTATTGTAAGTGTCGCACCATCAATGAGAATTTACAGAAATTCGCTGTGTCGTAATT 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 ATTCTATTGTAAGTGTCGCACCATCAATGAGAATTTACAGAAATTCGCTGTGTCGTAATT 780
Qy 781 GTGTTTCGTATGAGTTGCAGACTTCTGTATATGTTTCAAGGAAGAAGAAAACTATTCACT 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 GTGTTTCGTATGAGTTGCAGACTTCTGTATATGTTTCAAGGAAGAAGAAAACTATTCACT 840
Qy 841 ATGCCACGTGTTCACATAGAAATGGTTTATATTATTTTTTATTACTCCGCGTAGCGGTAT 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 ATGCCACGTGTTCACATAGAAATGGTTTATATTATTTTTTATTACTCCGCGTAGCGGTAT 900
Qy 901 GTTTGGGCTTTTAAGCCCAATTGGGGTTAGGCCCAATTCAGTAGCCAATGAGGAAGGCTT 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 GTTTGGGCTTTTAAGCCCAATTGGGGTTAGGCCCAATTCAGTAGCCAATGAGGAAGGCTT 960
Qy 961 AATGACCTCATATGATCGAGGTGCTGAG 988
||||||||||||||||||||||||||||
Db 961 AATGACCTCATATGATCGAGGTGCTGAG 988
Lal et al. also describes DNA isolation and PCR analysis using primers to amplify different DNA segments of nanovirus genome (p. 1027, right column, para 2). Primers are also used to detect presence of different genomic segments including DNA-M of the MDV genome (p. 1029; left column, last para). Moreover, designing a PCR primer is a well-known and routine method in the art (Addgene protocol). Lal et al. also describes AccuPower PCR Master Mix (Bioneer) (p. 1027, right column, para 2, line 21), which contains all the necessary reagents for the PCR including “Top DNA Polymerase, dNTPs, reaction buffer, tracking dye, and patented stabilizer” (Bioneer flyer).
However, Lal et al. does not describe instant SEQ ID NOs: 3-4.
Zhang et al. describes a (DNA-N) nucleotide sequence, BDP1, (page 2437, right column para 1, line 18; Suppl. Table S1; GenBank Accession No. LC094150) encoding a nuclear shuttle protein having 100% sequence identity to instant SEQ ID NO: 3, as shown below.
RESULT 1
LC094150
LOCUS LC094150 973 bp DNA linear VRL 12-MAY-2016
DEFINITION Milk vetch dwarf virus gene for nuclear shuttle protein, complete
cds, isolate: BDP1.
ACCESSION LC094150 (mentioned in Spec, page 25, line 12)
VERSION LC094150.1
SOURCE Milk vetch dwarf virus
ORGANISM Milk vetch dwarf virus
Viruses; Monodnaviria; Shotokuvirae; Cressdnaviricota;
Arfiviricetes; Mulpavirales; Nanoviridae; Nanovirus.
REFERENCE 1
AUTHORS Sano,Y. and Kamal,M.U.
TITLE Identification of milkvetch dwarf virus in Bangladesh
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 973)
AUTHORS Sano,Y.
TITLE Direct Submission
JOURNAL Submitted (31-OCT-2015) Contact:Yoshitaka Sano Faculty of
Agriculture, Niigata University, Agrobiology; 2-8050 Ikarashi,
Niigata city, Niigata Prefecture 950-2181, Japan URL
:http://www.agr.niigata-u.ac.jp/index.html
FEATURES Location/Qualifiers
source 1..973
/organism="Milk vetch dwarf virus"
/mol_type="genomic DNA"
/isolate="BDP1"
/host="Carica papaya"
/db_xref="taxon:67585"
/segment="DNA-N"
/geo_loc_name="Bangladesh"
/collection_date="2014-06-03"
CDS 340..801
/codon_start=1
/product="nuclear shuttle protein"
/protein_id="BAV00125.1"
/translation="MADWFASPLKTCTHVCDFPTLAGNPSQEITCCDSMKNKLNDSRK
VLLVSCSVSFNGSFYGGNRNVRGQLQISMLEDDGVCRPIGYVPIGGYLYHNDYGYYEG
EKTFNLDIESQYLKKDEDYNRKFIVSVLNENGLDSLCDLKVFIVHALRIKV
Query Match 100.0%; Score 973; Length 973;
Best Local Similarity 100.0%;
Matches 973; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 ACCCCCGCCCCAGGATCAGCGGAGTCATTTAGACTCGCTATAAGCCGTTAGATGTGTAGA 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 ACCCCCGCCCCAGGATCAGCGGAGTCATTTAGACTCGCTATAAGCCGTTAGATGTGTAGA 60
Qy 61 CACGTGTACAATCAGGATCTGTGATTCGTGAAGCGAATCTGACGGAAGATTGTACAATAC 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 CACGTGTACAATCAGGATCTGTGATTCGTGAAGCGAATCTGACGGAAGATTGTACAATAC 120
Qy 121 GCTAAACTATATATGTACGCGTGTTTAATTGCCATTGGTCCATATAGTAGTGGACATCAC 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 GCTAAACTATATATGTACGCGTGTTTAATTGCCATTGGTCCATATAGTAGTGGACATCAC 180
Qy 181 GCGTTTTCTTTTAACGCGGTTTACTTTATAAGTGGGGACCAGTAGTTGCTTTGTTTGCAC 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 GCGTTTTCTTTTAACGCGGTTTACTTTATAAGTGGGGACCAGTAGTTGCTTTGTTTGCAC 240
Qy 241 GAAAGGTTAATCTTTTGTGTTGAAGAAGCTTCTTCGTTGCTCTATAAATACAAGCTTCTT 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 GAAAGGTTAATCTTTTGTGTTGAAGAAGCTTCTTCGTTGCTCTATAAATACAAGCTTCTT 300
Qy 301 CTTCATCCTCTTCGTCATCATTTTCTGCAACTGCAAAAAATGGCAGATTGGTTCGCGAGT 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 CTTCATCCTCTTCGTCATCATTTTCTGCAACTGCAAAAAATGGCAGATTGGTTCGCGAGT 360
Qy 361 CCTTTGAAGACATGTACACATGTCTGCGATTTTCCAACCCTTGCTGGTAATCCATCACAA 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 CCTTTGAAGACATGTACACATGTCTGCGATTTTCCAACCCTTGCTGGTAATCCATCACAA 420
Qy 421 GAGATTACTTGCTGTGATAGTATGAAGAATAAGTTAAATGATTCGAGGAAGGTTTTACTT 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 GAGATTACTTGCTGTGATAGTATGAAGAATAAGTTAAATGATTCGAGGAAGGTTTTACTT 480
Qy 481 GTGAGTTGCAGTGTGAGCTTCAATGGTAGTTTCTATGGAGGTAACAGAAACGTTCGTGGT 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 GTGAGTTGCAGTGTGAGCTTCAATGGTAGTTTCTATGGAGGTAACAGAAACGTTCGTGGT 540
Qy 541 CAGTTGCAGATTTCTATGTTAGAAGACGACGGTGTTTGTAGACCAATTGGTTATGTCCCA 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 CAGTTGCAGATTTCTATGTTAGAAGACGACGGTGTTTGTAGACCAATTGGTTATGTCCCA 600
Qy 601 ATTGGTGGTTACTTGTACCATAACGACTATGGATATTACGAAGGTGAGAAGACATTCAAC 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 ATTGGTGGTTACTTGTACCATAACGACTATGGATATTACGAAGGTGAGAAGACATTCAAC 660
Qy 661 CTTGATATTGAATCCCAGTACTTGAAGAAGGACGAAGACTACAATAGGAAGTTTATTGTT 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 CTTGATATTGAATCCCAGTACTTGAAGAAGGACGAAGACTACAATAGGAAGTTTATTGTT 720
Qy 721 TCAGTTCTCAACGAGAATGGTTTAGACAGTTTATGCGACTTAAAGGTTTTTATTGTTCAT 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 TCAGTTCTCAACGAGAATGGTTTAGACAGTTTATGCGACTTAAAGGTTTTTATTGTTCAT 780
Qy 781 GCTCTTAGGATCAAAGTGTAATAATCATAATTATTGTAAGATTATGCAATTGAATAATAA 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 GCTCTTAGGATCAAAGTGTAATAATCATAATTATTGTAAGATTATGCAATTGAATAATAA 840
Qy 841 TATCAGGATGAATAATTTATTTTTAAATACTCTGCGTAGCTGTATGTTAGGCCCAATAGG 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 TATCAGGATGAATAATTTATTTTTAAATACTCTGCGTAGCTGTATGTTAGGCCCAATAGG 900
Qy 901 CCCATTTGTTAGTAAGCCCAAATGAATATGACGTCATCTGATCCCTTGCTGAGCTGGGGC 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 CCCATTTGTTAGTAAGCCCAAATGAATATGACGTCATCTGATCCCTTGCTGAGCTGGGGC 960
Qy 961 GGGGCTTAGTATT 973
|||||||||||||
Db 961 GGGGCTTAGTATT 973
Zhang et al. also describes a nucleotide sequence encoding DNA-U1 component of a Nanovirus (MDV) (GenBank accession No. KY070231; page 2440, left column, para 2) comprising 100% sequence identity to instant SEQ ID NO: 4, as shown below.
RESULT 1
KY070231
LOCUS KY070231 990 bp DNA circular VRL 12-APR-2017
DEFINITION Milk vetch dwarf virus isolate VF segment DNA-U1, complete
sequence.
ACCESSION KY070231 (From Broad bean, V. faba)
VERSION KY070231.1
SOURCE Milk vetch dwarf virus
ORGANISM Milk vetch dwarf virus
Viruses; Monodnaviria; Shotokuvirae; Cressdnaviricota;
Arfiviricetes; Mulpavirales; Nanoviridae; Nanovirus.
REFERENCE 1 (bases 1 to 990)
AUTHORS Zhang,C., Zheng,H., Yan,D., Han,K., Song,X., Liu,Y., Zhang,D.,
Chen,J. and Yan,F.
TITLE Complete genomic characterization of milk vetch dwarf virus
isolates from cowpea and broad bean in Anhui province, China
JOURNAL Arch. Virol. (2017) In press
PUBMED 28386650
REMARK Publication Status: Available-Online prior to print
REFERENCE 2 (bases 1 to 990)
AUTHORS Zhang,C., Zheng,H., Yan,D., Han,K., Song,X., Liu,Y., Zhang,D.,
Chen,J. and Yan,F.
TITLE Direct Submission
JOURNAL Submitted (01-NOV-2016) ICTV, 24 Woodland Way, Stevenage,
Hertfordshire SG2 8BT, United Kingdom
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..990
/organism="Milk vetch dwarf virus"
/mol_type="genomic DNA"
/isolate="VF"
/host="broad bean"
/db_xref="taxon:67585"
/segment="DNA-U1"
/geo_loc_name="China: Anhui, Hefei City"
stem_loop 1..29
regulatory 274..280
/regulatory_class="TATA_box"
regulatory 372..824
/regulatory_class="polyA_signal_sequence"
regulatory 823..828
/regulatory_class="polyA_signal_sequence
Query Match 100.0%; Score 990; Length 990; Best Local Similarity 100.0%;
Matches 990; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 GGGGCGGGGCTTAGTATTACCCCCGCCCCGGGTTCAGAGTCACGTACGGAGTGACTTACA 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 GGGGCGGGGCTTAGTATTACCCCCGCCCCGGGTTCAGAGTCACGTACGGAGTGACTTACA 60
Qy 61 GCCGTTGGATTTTAAATAAGATGGACGATCAGGATCTGTGATTCGTGAAGCGAATCTGAC 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 GCCGTTGGATTTTAAATAAGATGGACGATCAGGATCTGTGATTCGTGAAGCGAATCTGAC 120
Qy 121 GGAAGATCGTCCGAAGCTTCGTGGTAGGGCCCCCCTATTGCTTTATCTTTACTTTAATAA 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 GGAAGATCGTCCGAAGCTTCGTGGTAGGGCCCCCCTATTGCTTTATCTTTACTTTAATAA 180
Qy 181 AGTAAAGTAAGATGCTGTCCCCTACTTTATTCGTTTATGAGTTGTTTACAGCTGTCTTTG 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 AGTAAAGTAAGATGCTGTCCCCTACTTTATTCGTTTATGAGTTGTTTACAGCTGTCTTTG 240
Qy 241 CTTCGTCTCGAAGCAAAGGACCTTCTCATCGTCTATAAAAGCTGTGTTTATGAAGAAGAT 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 CTTCGTCTCGAAGCAAAGGACCTTCTCATCGTCTATAAAAGCTGTGTTTATGAAGAAGAT 300
Qy 301 GAATTGTTCATCTTCTTCTTTCTCACAAACCATATTGTTTGTTTCATCAAACTAAAGCTT 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 GAATTGTTCATCTTCTTCTTTCTCACAAACCATATTGTTTGTTTCATCAAACTAAAGCTT 360
Qy 361 CTATTGAGGTTATGTCTGCGAACGATTCTTCGTTGTTTGATGAGGCCACCGATGAGTTAA 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 CTATTGAGGTTATGTCTGCGAACGATTCTTCGTTGTTTGATGAGGCCACCGATGAGTTAA 420
Qy 421 TATCGTCTGAGAGGAAATTGATAGCCGTCGAATGCCATGATGATGATTCTCAGGTAATTA 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 TATCGTCTGAGAGGAAATTGATAGCCGTCGAATGCCATGATGATGATTCTCAGGTAATTA 480
Qy 481 ATGTTAAGGTTGAAGATATCTTTTGCGATATGTCCGACAAGGTCGTTATTAAACTGCAGT 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 ATGTTAAGGTTGAAGATATCTTTTGCGATATGTCCGACAAGGTCGTTATTAAACTGCAGT 540
Qy 541 TTAGATTGTGTTACAAGTACCGAAAGCTATTAGATATTACTCTTCTTGGGTGTCGTATGA 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 TTAGATTGTGTTACAAGTACCGAAAGCTATTAGATATTACTCTTCTTGGGTGTCGTATGA 600
Qy 601 AGGTTCATACAGAGTTGAAGACACCATCTGAACGTTCTCTGAAGAGTATTTTGCAGAAGA 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 AGGTTCATACAGAGTTGAAGACACCATCTGAACGTTCTCTGAAGAGTATTTTGCAGAAGA 660
Qy 661 AGATGAATGTTATATGCGATGGGAATTATTTGATAGGTATTAGAATGTTTTTTATAAATA 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 AGATGAATGTTATATGCGATGGGAATTATTTGATAGGTATTAGAATGTTTTTTATAAATA 720
Qy 721 TTAATCAGTTGATTAATACTTGTAAGTGGATTACCAGAACTGAAGATGTATATCCAATAT 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 TTAATCAGTTGATTAATACTTGTAAGTGGATTACCAGAACTGAAGATGTATATCCAATAT 780
Qy 781 GTACTTTGTATCATGTGAATGATATTCCTGTAATTGATAGTTAATAATGAATATTTGTTT 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 GTACTTTGTATCATGTGAATGATATTCCTGTAATTGATAGTTAATAATGAATATTTGTTT 840
Qy 841 TTTGATCAAACACATGTATTGATAAATATATTATTTTTAATTACTCTGCGAAGCTATATG 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 TTTGATCAAACACATGTATTGATAAATATATTATTTTTAATTACTCTGCGAAGCTATATG 900
Qy 901 TCTAGGCCCAATAGGCCCAAATGCTCTTAAGCCCAATGAAATTAACACTGAAGTGAGTCA 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 TCTAGGCCCAATAGGCCCAAATGCTCTTAAGCCCAATGAAATTAACACTGAAGTGAGTCA 960
Qy 961 GCTGACGTCAGCTGACTCCTTGATGACGTA 990
||||||||||||||||||||||||||||||
Db 961 GCTGACGTCAGCTGACTCCTTGATGACGTA 990
Zhang et al. also describes primers to amplify and detect presence of a nanovirus nucleotide sequences (page 2437, right column, last para; and page 2438, left column, first para; Suppl. Table S2).
Before the effective filling date, it would have been obvious to an ordinarily skilled artisan to use the composition comprising suitable primers along with other required reagents to diagnose/detect the presence or absence of a polynucleotide sequence of the nanoviral genome as described by Lal et al. and Zhang et al.
Before the effective filling of the invention, an ordinarily skilled artisan would have been motivated to use a composition comprising specific primers and other reagents to diagnose or detect the presence or absence of any specific genomic nucleotide sequence of a nanovirus.
Regarding claim 8, Lal et al. (page 1027, right column, para 2, line 12-18) and Zhang et al. (page 2437, right column last para; p 2438, left column, first para; Suppln. Table S2) describe primers that can detect DNA-R sequence. Lal et al. describes a DNA-R sequence comprising SEQ ID NO: 9, as shown below.
RESULT 1
MK726377
LOCUS MK726377 1002 bp DNA linear VRL 01-MAY-2020
DEFINITION Milk vetch dwarf virus isolate YS-AA-1 segment R, complete
sequence.
ACCESSION MK726377
VERSION MK726377.1
SOURCE Milk vetch dwarf virus
ORGANISM Milk vetch dwarf virus
Viruses; Monodnaviria; Shotokuvirae; Cressdnaviricota;
Arfiviricetes; Mulpavirales; Nanoviridae; Nanovirus.
REFERENCE 1 (bases 1 to 1002)
AUTHORS Lal,A., Kil,E.-J., Byun,H.-S., Zarghami,S.D., Kim,J.-K. and Lee,S.
TITLE Milk vetch dwarf virus R segment isolate Korea
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 1002)
AUTHORS Lal,A., Kil,E.-J., Byun,H.-S., Zarghami,S.D., Kim,J.-K. and Lee,S.
TITLE Direct Submission
JOURNAL Submitted (29-MAR-2019) Department of Integrative Biotechnology,
Sungkyunkwan University, 2066, Seobu-ro, Jangan-gu, Suwon, Gyeonggi
16419, Korea
FEATURES Location/Qualifiers
source 1..1002
/organism="Milk vetch dwarf virus"
/mol_type="genomic DNA"
/isolate="YS-AA-1"
/host="Carica papaya"
/db_xref="taxon:67585"
/segment="R"
/geo_loc_name="South Korea"
/collection_date="Sep-2017"
CDS 120..980
/note="Rep"
/codon_start=1
/product="Rep Protein"
/protein_id="QHA79702.1"
/translation="MARQVICWCFTLNNPLSPLVLHESMKYLVYQREQGEAGNIHFQG
YIEMKKRTSLAGMKRLVPGAHFEKMRVNQDQAKAYAMKEDTRLEGPWEYGEFIPTIED
KLREVMNDMKITGKRPIEYIEECCNTYDKSASTLREFRGELKKKKAILSWELQRKPWM
DEVDTLLQERDGRRIIWVYGPQGGEGKTSYAKHLVKTRDAFYSTGGKTADIAFAWDHQ
ELVLFDFPRSFEEYVNYGVIEQLKNGIIQSGKYQSVIKYSDYVEVIVFANFTPRSGMF
SDDRIVYVYA
Query Match 100.0%; Score 1002; Length 1002;
Best Local Similarity 100.0%;
Matches 1002; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 CTGGGGCGGGGCTTAGTATTACCCCCGCCCCAGGATCAGCGGAGTCATCACGTGACCCGC 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 CTGGGGCGGGGCTTAGTATTACCCCCGCCCCAGGATCAGCGGAGTCATCACGTGACCCGC 60
Qy 61 ACATGCCTTTAGTCTATATATAGCCCAAGTTGTAATATTTTATCATTCATCAATAAAATA 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 ACATGCCTTTAGTCTATATATAGCCCAAGTTGTAATATTTTATCATTCATCAATAAAATA 120
Qy 121 TGGCTCGGCAAGTTATATGCTGGTGCTTTACATTAAATAATCCTCTCTCTCCTCTAGTTC 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 TGGCTCGGCAAGTTATATGCTGGTGCTTTACATTAAATAATCCTCTCTCTCCTCTAGTTC 180
Qy 181 TTCATGAATCAATGAAGTACCTTGTTTATCAAAGAGAACAAGGTGAAGCTGGAAATATTC 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 TTCATGAATCAATGAAGTACCTTGTTTATCAAAGAGAACAAGGTGAAGCTGGAAATATTC 240
Qy 241 ATTTCCAGGGTTATATTGAGATGAAGAAACGCACGTCTCTTGCAGGTATGAAGCGATTAG 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 ATTTCCAGGGTTATATTGAGATGAAGAAACGCACGTCTCTTGCAGGTATGAAGCGATTAG 300
Qy 301 TTCCAGGAGCCCATTTTGAGAAAATGAGGGTAAACCAAGACCAGGCTAAGGCTTACGCTA 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 TTCCAGGAGCCCATTTTGAGAAAATGAGGGTAAACCAAGACCAGGCTAAGGCTTACGCTA 360
Qy 361 TGAAGGAAGACACCCGTCTTGAAGGTCCATGGGAGTATGGGGAGTTCATCCCCACCATTG 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 TGAAGGAAGACACCCGTCTTGAAGGTCCATGGGAGTATGGGGAGTTCATCCCCACCATTG 420
Qy 421 AAGATAAGCTCAGGGAAGTTATGAACGACATGAAGATTACAGGTAAGAGACCTATTGAGT 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 AAGATAAGCTCAGGGAAGTTATGAACGACATGAAGATTACAGGTAAGAGACCTATTGAGT 480
Qy 481 ATATAGAAGAGTGCTGTAATACATACGATAAATCTGCAAGTACTCTAAGGGAGTTCAGAG 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 ATATAGAAGAGTGCTGTAATACATACGATAAATCTGCAAGTACTCTAAGGGAGTTCAGAG 540
Qy 541 GTGAGTTGAAGAAGAAGAAGGCAATTTTAAGTTGGGAGTTGCAGAGGAAGCCATGGATGG 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 GTGAGTTGAAGAAGAAGAAGGCAATTTTAAGTTGGGAGTTGCAGAGGAAGCCATGGATGG 600
Qy 601 ACGAGGTCGATACCTTGCTTCAGGAGAGAGATGGAAGAAGAATCATTTGGGTGTATGGCC 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 ACGAGGTCGATACCTTGCTTCAGGAGAGAGATGGAAGAAGAATCATTTGGGTGTATGGCC 660
Qy 661 CACAAGGTGGAGAAGGGAAAACCTCTTACGCTAAGCATCTTGTGAAAACGCGTGATGCTT 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 CACAAGGTGGAGAAGGGAAAACCTCTTACGCTAAGCATCTTGTGAAAACGCGTGATGCTT 720
Qy 721 TTTATTCGACAGGTGGAAAGACAGCCGACATTGCTTTTGCGTGGGACCACCAAGAGTTAG 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 TTTATTCGACAGGTGGAAAGACAGCCGACATTGCTTTTGCGTGGGACCACCAAGAGTTAG 780
Qy 781 TGCTTTTCGACTTTCCTCGAAGCTTCGAGGAGTATGTTAATTATGGAGTAATAGAGCAAT 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 TGCTTTTCGACTTTCCTCGAAGCTTCGAGGAGTATGTTAATTATGGAGTAATAGAGCAAT 840
Qy 841 TAAAGAATGGGATTATCCAGTCTGGCAAGTACCAAAGTGTAATTAAGTATAGTGATTATG 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 TAAAGAATGGGATTATCCAGTCTGGCAAGTACCAAAGTGTAATTAAGTATAGTGATTATG 900
Qy 901 TGGAGGTGATTGTATTTGCTAATTTTACTCCGCGTAGCGGTATGTTTAGTGATGATAGGA 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 TGGAGGTGATTGTATTTGCTAATTTTACTCCGCGTAGCGGTATGTTTAGTGATGATAGGA 960
Qy 961 TTGTATATGTATATGCATGACGTCATATGATCCCGTGCTGAG 1002
||||||||||||||||||||||||||||||||||||||||||
Db 961 TTGTATATGTATATGCATGACGTCATATGATCCCGTGCTGAG 1002
Regarding claim 9, Lal et al. describes the detecting agents comprising primers that specifically bind to nanovirus DNA-M (page 1027, right column, para 2, line 4) and DNA-C (page 1027, right column, para 2, line 13), and as described above.
Regarding claim 10, Lal et al. describes primers that specifically bind to DNA-R and DNA-S (p. 1027, right column, para 2, line 4 and 13).
Regarding claims 11-12, Lal et al. (p. 1027, right column, para 2) and Zhang et al. ((page 2437, right column, last para; and page 2438, left column, first para; Suppl. Table S2) describe a method, PCR, that can detect Nanovirus infection in terms of detecting presence or absence of SEQ ID NOs: 1-4, in plant derived samples, as described above. The method comprises contacting the diagnostic composition with a plant-derived sample and performing a nucleic acid amplification reaction; and detecting amplified nucleic acid.
Regarding claim 13, Lal et al. (page 1027, right column, para 2, line 12-18) and Zhang et al. (page 2437, right column last para; p 2438, left column, first para; Suppln. Table S2) describe a method to detect DNA-R sequences and Lal et al. describes a DNA-R nucleotide sequence comprising SEQ ID NO: 9, as shown above.
Conclusion
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