A method for identifying brucella abortus based on specific probe and SNP site

Abstract

The application discloses a method for identifying bovine brucella based on specific probes and SNP sites. The application relates to the technical field of biology, and particularly relates to a method for identifying bovine brucella based on specific probes and SNP sites. The method for identifying or assisting in identifying bovine brucella comprises the following steps: S1), extracting genomic DNA (double-stranded DNA) of a sample to be detected; S2), sequencing the genomic DNA to obtain a sequencing result; S3), performing sequence alignment on the sequencing result and a specific probe for identifying bovine brucella, and determining or assisting in determining whether the sample to be detected is bovine brucella or contains bovine brucella according to the sequence alignment result. The method can quickly, simply and efficiently identify whether the sample contains bovine brucella.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to a method for identifying Brucella bovis based on specific probes and SNP sites. Background Technology

[0002] Brucella is a facultative intracellular parasitic Gram-negative bacterium that causes brucellosis, a chronic infectious disease that can be transmitted between humans and animals. Brucellosis can cause abortions in livestock such as cattle, pigs, and sheep, and high fever in humans. It damages the reproductive systems and joints of both animals and humans, posing a significant threat to livestock development, human health, and public health. It is the most widespread zoonotic disease in the world. Human infection with brucellosis primarily occurs through contact with infected livestock or by consuming food or animal products contaminated with the bacterium. Brucellosis is classified as a Class B infectious disease under the Law of the People's Republic of China on the Prevention and Control of Infectious Diseases. Due to its high infectivity, Brucella is considered a potential biological weapon.

[0003] Based on the biochemical characteristics, selectivity to natural hosts, and pathogenicity of Brucella, the genus Brucella is mainly divided into six species: *Brucella melitensis*, *Brucella abortus*, *Brucella suis*, *Brucella neotomae*, *Brucella canis*, and *Brucella ovis*. Traditional methods of Brucella species identification rely on phenotypic techniques and the initial host of isolation, which are labor-intensive and slow. Molecular biological methods, on the other hand, utilize genetic markers for differentiation, offering simplicity and speed. Currently, a single primer pair is insufficient to distinguish between different species of Brucella; multiplex PCR is required for species identification.

[0004] With the development of sequencing technology, more and more Brucella species have obtained their whole genome sequences. From the perspective of comparative genomics, structural differences and similarity analyses are performed on Brucella species across the entire genome to screen for specific fragments, providing specific sequences for species identification and thus better distinguishing different Brucella species. Summary of the Invention

[0005] The main problem this invention aims to solve is how to identify or assist in the identification of Brucella bovis.

[0006] To address the aforementioned problems, this invention provides a method for identifying or assisting in the identification of Brucella bovis.

[0007] The present invention provides a method for identifying or assisting in the identification of Brucella bovis, the method comprising the following steps:

[0008] S1) Extract genomic DNA (double-stranded DNA) from the sample to be tested;

[0009] S2) Sequencing the genomic DNA to obtain sequencing results;

[0010] S3) The sequencing results are compared with the specific probe for identifying Brucella bovis. Based on the sequence comparison results, it is determined or assisted in determining whether the sample to be tested is Brucella bovis or whether it contains Brucella bovis.

[0011] If the sequence alignment results show that the genomic DNA sequence of the sample to be tested contains any of the specific probes, the sample to be tested is Brucella bovis or contains Brucella bovis; if the sequence alignment results show that the genomic DNA sequence of the sample to be tested does not contain the specific probes, the sample to be tested is not Brucella bovis or does not contain Brucella bovis.

[0012] The specific probe contains at least one of 28 single-stranded DNAs named probe 1 to probe 28:

[0013] Probe 1 contains SNP (single nucleotide polymorphism) site 1, which is the 100th nucleotide of sequence 1 and is T;

[0014] Probe 2 contains SNP (single nucleotide polymorphism) site 2, which is the 100th nucleotide of sequence 2 and is T;

[0015] Probe 3 contains SNP (single nucleotide polymorphism) site 3, which is the 100th nucleotide of sequence 3 and is T;

[0016] Probe 4 contains SNP (single nucleotide polymorphism) site 4, which is the 100th nucleotide of sequence 4 and is A;

[0017] Probe 5 contains SNP (single nucleotide polymorphism) site 5, which is the 100th nucleotide of sequence 5 and is T;

[0018] Probe 6 contains SNP (single nucleotide polymorphism) site 6, which is the 92nd nucleotide of sequence 6 and is C.

[0019] Probe 7 contains SNP (single nucleotide polymorphism) site 7, which is the 63rd nucleotide of sequence 7 and is C.

[0020] Probe 8 contains SNP (single nucleotide polymorphism) site 8, which is the 100th nucleotide of sequence 8 and is C.

[0021] Probe 9 contains SNP (single nucleotide polymorphism) site 9, which is the 68th nucleotide of sequence 9 and is A;

[0022] Probe 10 contains an SNP (single nucleotide polymorphism) site 10, which is the 79th nucleotide of sequence 10 and is A.

[0023] Probe 11 contains an SNP (single nucleotide polymorphism) site 11, which is the 74th nucleotide of sequence 11 and is C.

[0024] Probe 12 contains an SNP (single nucleotide polymorphism) site 12, which is the 87th nucleotide of sequence 12 and is T.

[0025] Probe 13 contains an SNP (single nucleotide polymorphism) site 13, wherein the SNP site 13 is the 63rd nucleotide of sequence 13 and the nucleotide is G, and the SNP site 14 is the 64th nucleotide of sequence 13 and the nucleotide is C.

[0026] Probe 14 contains an SNP (single nucleotide polymorphism) site 15, which is the 100th nucleotide of sequence 14 and is G.

[0027] Probe 15 contains an SNP (single nucleotide polymorphism) site 16, which is the 69th nucleotide of sequence 15 and is nucleotide A;

[0028] Probe 16 contains an SNP (single nucleotide polymorphism) site 17, which is the 20th nucleotide of sequence 16 and is C; and an SNP site 18, which is the 151st nucleotide of sequence 16 and is G.

[0029] Probe 17 contains SNP (single nucleotide polymorphism) site 19, which is the 81st nucleotide of sequence 17 and is T;

[0030] Probe 18 contains an SNP (single nucleotide polymorphism) site 20, which is the 90th nucleotide of sequence 18 and is nucleotide A;

[0031] Probe 19 contains an SNP (single nucleotide polymorphism) site 21, which is the 99th nucleotide of sequence 19 and is nucleotide A;

[0032] Probe 20 contains an SNP (single nucleotide polymorphism) site 22, which is the 92nd nucleotide of sequence 20 and is T.

[0033] Probe 21 contains an SNP (single nucleotide polymorphism) site 23, which is the 71st nucleotide of sequence 21 and is nucleotide A;

[0034] Probe 22 contains an SNP (single nucleotide polymorphism) site 24, which is the 100th nucleotide of sequence 22 and is T.

[0035] Probe 23 contains an SNP (single nucleotide polymorphism) site 25, which is the 100th nucleotide of sequence 23 and is T.

[0036] Probe 24 contains an SNP (single nucleotide polymorphism) site 26, which is the 58th nucleotide of sequence 24 and is T.

[0037] Probe 25 contains an SNP (single nucleotide polymorphism) site 27, which is the 100th nucleotide of sequence 25 and is A, and an SNP site 28, which is the 136th nucleotide of sequence 25 and is T.

[0038] Probe 26 contains SNP (single nucleotide polymorphism) site 29, which is the 78th nucleotide of sequence 26 and is T;

[0039] Probe 27 contains an SNP (single nucleotide polymorphism) site 30, which is the 100th nucleotide of sequence 27 and is T.

[0040] Probe 28 contains an SNP (single nucleotide polymorphism) site 31, which is the 69th nucleotide of sequence 28 and is T.

[0041] In the above method, the nucleotide sequence of probe 1 is sequence 1 and the nucleotide of SNP site 1 is T;

[0042] The nucleotide sequence of probe 2 is sequence 2 and the nucleotide of SNP site 2 is T;

[0043] The nucleotide sequence of probe 3 is sequence 3 and the nucleotide of SNP site 3 is T;

[0044] The nucleotide sequence of probe 4 is sequence 4 and the nucleotide of SNP site 4 is A;

[0045] The nucleotide sequence of probe 5 is sequence 5 and the nucleotide of SNP site 5 is T;

[0046] The nucleotide sequence of probe 6 is sequence 6 and the nucleotide of SNP site 6 is C;

[0047] The nucleotide sequence of probe 7 is sequence 7 and the nucleotide of SNP site 7 is C;

[0048] The nucleotide sequence of probe 8 is sequence 8 and the nucleotide of SNP site 8 is C;

[0049] The nucleotide sequence of probe 9 is sequence 9 and the nucleotide of SNP site 9 is A;

[0050] The nucleotide sequence of probe 10 is sequence 10 and the nucleotide of SNP site 10 is A;

[0051] The nucleotide sequence of probe 11 is sequence 11 and the nucleotide of SNP site 11 is C;

[0052] The nucleotide sequence of probe 12 is sequence 12 and the nucleotide of SNP site 12 is T;

[0053] The nucleotide sequence of probe 13 is sequence 13, the nucleotide of SNP site 13 is G, and the nucleotide of SNP site 14 is C.

[0054] The nucleotide sequence of probe 14 is sequence 14 and the nucleotide of SNP site 15 is G;

[0055] The nucleotide sequence of probe 15 is sequence 15 and the nucleotide of SNP site 16 is A;

[0056] The nucleotide sequence of probe 16 is sequence 16, and the nucleotide of SNP site 17 is C; and the nucleotide of SNP site 18 is G;

[0057] The nucleotide sequence of probe 17 is sequence 17 and the nucleotide of SNP site 19 is T;

[0058] The nucleotide sequence of probe 18 is sequence 18 and the nucleotide of SNP site 20 is A;

[0059] The nucleotide sequence of probe 19 is sequence 19 and the nucleotide of SNP site 21 is A;

[0060] The nucleotide sequence of probe 20 is sequence 20 and the nucleotide of SNP site 22 is T;

[0061] The nucleotide sequence of probe 21 is sequence 21 and the nucleotide of SNP site 23 is A;

[0062] The nucleotide sequence of probe 22 is sequence 22 and the nucleotide of SNP site 24 is T;

[0063] The nucleotide sequence of probe 23 is sequence 23 and the nucleotide of SNP site 25 is T;

[0064] The nucleotide sequence of probe 24 is sequence 24 and the nucleotide of SNP site 26 is T;

[0065] The nucleotide sequence of probe 25 is sequence 25, the nucleotide of SNP site 27 is A, and the nucleotide of SNP site 28 is T.

[0066] The nucleotide sequence of probe 26 is sequence 26 and the nucleotide of SNP site 29 is T;

[0067] The nucleotide sequence of probe 27 is sequence 27 and the nucleotide of SNP site 30 is T;

[0068] The nucleotide sequence of probe 28 is sequence 28 and the nucleotide of SNP site 31 is T.

[0069] The sample to be tested can be a sample from a non-living human or animal body, such as an environmental sample (e.g., air) or a food sample (e.g., frozen or fresh food).

[0070] The present invention also provides an apparatus for identifying or assisting in the identification of Brucella bovis, the apparatus comprising:

[0071] 1) Sequence alignment module: Aligns the genomic DNA of the sample to be tested with specific probes for identifying Brucella bovis, and outputs the sequence alignment results. The specific probes contain at least one of 28 single-stranded DNAs named probe 1 to probe 28.

[0072] Probe 1 contains SNP (single nucleotide polymorphism) site 1, which is the 100th nucleotide of sequence 1 and is T;

[0073] Probe 2 contains SNP (single nucleotide polymorphism) site 2, which is the 100th nucleotide of sequence 2 and is T;

[0074] Probe 3 contains SNP (single nucleotide polymorphism) site 3, which is the 100th nucleotide of sequence 3 and is T;

[0075] Probe 4 contains SNP (single nucleotide polymorphism) site 4, which is the 100th nucleotide of sequence 4 and is A;

[0076] Probe 5 contains SNP (single nucleotide polymorphism) site 5, which is the 100th nucleotide of sequence 5 and is T;

[0077] Probe 6 contains SNP (single nucleotide polymorphism) site 6, which is the 92nd nucleotide of sequence 6 and is C.

[0078] Probe 7 contains SNP (single nucleotide polymorphism) site 7, which is the 63rd nucleotide of sequence 7 and is C.

[0079] Probe 8 contains SNP (single nucleotide polymorphism) site 8, which is the 100th nucleotide of sequence 8 and is C.

[0080] Probe 9 contains SNP (single nucleotide polymorphism) site 9, which is the 68th nucleotide of sequence 9 and is A;

[0081] Probe 10 contains an SNP (single nucleotide polymorphism) site 10, which is the 79th nucleotide of sequence 10 and is A.

[0082] Probe 11 contains an SNP (single nucleotide polymorphism) site 11, which is the 74th nucleotide of sequence 11 and is C.

[0083] Probe 12 contains an SNP (single nucleotide polymorphism) site 12, which is the 87th nucleotide of sequence 12 and is T.

[0084] Probe 13 contains an SNP (single nucleotide polymorphism) site 13, wherein the SNP site 13 is the 63rd nucleotide of sequence 13 and the nucleotide is G, and the SNP site 14 is the 64th nucleotide of sequence 13 and the nucleotide is C.

[0085] Probe 14 contains an SNP (single nucleotide polymorphism) site 15, which is the 100th nucleotide of sequence 14 and is G.

[0086] Probe 15 contains an SNP (single nucleotide polymorphism) site 16, which is the 69th nucleotide of sequence 15 and is nucleotide A;

[0087] Probe 16 contains an SNP (single nucleotide polymorphism) site 17, which is the 20th nucleotide of sequence 16 and is C; and an SNP site 18, which is the 151st nucleotide of sequence 16 and is G.

[0088] Probe 17 contains SNP (single nucleotide polymorphism) site 19, which is the 81st nucleotide of sequence 17 and is T;

[0089] Probe 18 contains an SNP (single nucleotide polymorphism) site 20, which is the 90th nucleotide of sequence 18 and is nucleotide A;

[0090] Probe 19 contains an SNP (single nucleotide polymorphism) site 21, which is the 99th nucleotide of sequence 19 and is nucleotide A;

[0091] Probe 20 contains an SNP (single nucleotide polymorphism) site 22, which is the 92nd nucleotide of sequence 20 and is T.

[0092] Probe 21 contains an SNP (single nucleotide polymorphism) site 23, which is the 71st nucleotide of sequence 21 and is nucleotide A;

[0093] Probe 22 contains an SNP (single nucleotide polymorphism) site 24, which is the 100th nucleotide of sequence 22 and is T.

[0094] Probe 23 contains an SNP (single nucleotide polymorphism) site 25, which is the 100th nucleotide of sequence 23 and is T.

[0095] Probe 24 contains an SNP (single nucleotide polymorphism) site 26, which is the 58th nucleotide of sequence 24 and is T.

[0096] Probe 25 contains an SNP (single nucleotide polymorphism) site 27, which is the 100th nucleotide of sequence 25 and is A, and an SNP site 28, which is the 136th nucleotide of sequence 25 and is T.

[0097] Probe 26 contains SNP (single nucleotide polymorphism) site 29, which is the 78th nucleotide of sequence 26 and is T;

[0098] Probe 27 contains an SNP (single nucleotide polymorphism) site 30, which is the 100th nucleotide of sequence 27 and is T.

[0099] Probe 28 contains an SNP (single nucleotide polymorphism) site 31, which is the 69th nucleotide of sequence 28 and is T.

[0100] 2) Identification module: Based on the sequence alignment results, determine or assist in determining whether the sample to be tested is Brucella bovis or whether it contains Brucella bovis; if the sequence alignment results show that the genomic DNA sequence of the sample to be tested contains any one of the specific probes, the sample to be tested is Brucella bovis or contains Brucella bovis; if the sequence alignment results show that the genomic DNA sequence of the sample to be tested does not contain the specific probe, the sample to be tested is not Brucella bovis or does not contain Brucella bovis.

[0101] In the above-described apparatus, the nucleotide sequence of probe 1 is sequence 1, and the nucleotide of SNP site 1 is T.

[0102] The nucleotide sequence of probe 2 is sequence 2 and the nucleotide of SNP site 2 is T;

[0103] The nucleotide sequence of probe 3 is sequence 3 and the nucleotide of SNP site 3 is T;

[0104] The nucleotide sequence of probe 4 is sequence 4 and the nucleotide of SNP site 4 is A;

[0105] The nucleotide sequence of probe 5 is sequence 5 and the nucleotide of SNP site 5 is T;

[0106] The nucleotide sequence of probe 6 is sequence 6 and the nucleotide of SNP site 6 is C;

[0107] The nucleotide sequence of probe 7 is sequence 7 and the nucleotide of SNP site 7 is C;

[0108] The nucleotide sequence of probe 8 is sequence 8 and the nucleotide of SNP site 8 is C;

[0109] The nucleotide sequence of probe 9 is sequence 9 and the nucleotide of SNP site 9 is A;

[0110] The nucleotide sequence of probe 10 is sequence 10 and the nucleotide of SNP site 10 is A;

[0111] The nucleotide sequence of probe 11 is sequence 11 and the nucleotide of SNP site 11 is C;

[0112] The nucleotide sequence of probe 12 is sequence 12 and the nucleotide of SNP site 12 is T;

[0113] The nucleotide sequence of probe 13 is sequence 13, the nucleotide of SNP site 13 is G, and the nucleotide of SNP site 14 is C.

[0114] The nucleotide sequence of probe 14 is sequence 14 and the nucleotide of SNP site 15 is G;

[0115] The nucleotide sequence of probe 15 is sequence 15 and the nucleotide of SNP site 16 is A;

[0116] The nucleotide sequence of probe 16 is sequence 16, and the nucleotide of SNP site 17 is C; and the nucleotide of SNP site 18 is G;

[0117] The nucleotide sequence of probe 17 is sequence 17 and the nucleotide of SNP site 19 is T;

[0118] The nucleotide sequence of probe 18 is sequence 18 and the nucleotide of SNP site 20 is A;

[0119] The nucleotide sequence of probe 19 is sequence 19 and the nucleotide of SNP site 21 is A;

[0120] The nucleotide sequence of probe 20 is sequence 20 and the nucleotide of SNP site 22 is T;

[0121] The nucleotide sequence of probe 21 is sequence 21 and the nucleotide of SNP site 23 is A;

[0122] The nucleotide sequence of probe 22 is sequence 22 and the nucleotide of SNP site 24 is T;

[0123] The nucleotide sequence of probe 23 is sequence 23 and the nucleotide of SNP site 25 is T;

[0124] The nucleotide sequence of probe 24 is sequence 24 and the nucleotide of SNP site 26 is T;

[0125] The nucleotide sequence of probe 25 is sequence 25, the nucleotide of SNP site 27 is A, and the nucleotide of SNP site 28 is T.

[0126] The nucleotide sequence of probe 26 is sequence 26 and the nucleotide of SNP site 29 is T;

[0127] The nucleotide sequence of probe 27 is sequence 27 and the nucleotide of SNP site 30 is T;

[0128] The nucleotide sequence of probe 28 is sequence 28 and the nucleotide of SNP site 31 is T.

[0129] The present invention also provides a computer-readable storage medium storing a computer program that causes a computer to perform the steps described above.

[0130] The application of the device described above in the identification or auxiliary identification of Brucella bovis, or in the preparation of products for the identification or auxiliary identification of Brucella bovis, also falls within the scope of protection claimed by this invention.

[0131] The use of the computer-readable storage medium described above in the identification or assistance of Brucella bovis, or in the preparation of products for the identification or assistance of Brucella bovis, is also within the scope of protection claimed by this invention.

[0132] The present invention also provides a specific probe for identifying Brucella bovis, the specific probe comprising at least one of 28 single-stranded DNAs named probe 1 to probe 28, wherein probe 1 contains SNP (single nucleotide polymorphism) site 1, wherein SNP site 1 is the 100th nucleotide of sequence 1 and the nucleotide is T.

[0133] The probe 2 contains SNP (single nucleotide polymorphism) site 2, which is the 100th nucleotide of sequence 2 and is T.

[0134] The probe 3 contains an SNP (single nucleotide polymorphism) site 3, which is the 100th nucleotide of sequence 3 and is T.

[0135] The probe 4 contains a SNP (single nucleotide polymorphism) site 4, which is the 100th nucleotide of sequence 4 and is A.

[0136] The probe 5 contains an SNP (single nucleotide polymorphism) site 5, which is the 100th nucleotide of sequence 5 and is T.

[0137] The probe 6 contains an SNP (single nucleotide polymorphism) site 6, which is the 92nd nucleotide of sequence 6 and is C.

[0138] The probe 7 contains an SNP (single nucleotide polymorphism) site 7, which is the 63rd nucleotide of sequence 7 and is C.

[0139] The probe 8 contains a SNP (single nucleotide polymorphism) site 8, which is the 100th nucleotide of sequence 8 and is C.

[0140] The probe 9 contains a SNP (single nucleotide polymorphism) site 9, which is the 68th nucleotide of sequence 9 and is A.

[0141] The probe 10 contains an SNP (single nucleotide polymorphism) site 10, which is the 79th nucleotide of sequence 10 and is A.

[0142] The probe 11 contains an SNP (single nucleotide polymorphism) site 11, which is the 74th nucleotide of sequence 11 and is C.

[0143] The probe 12 contains an SNP (single nucleotide polymorphism) site 12, which is the 87th nucleotide of sequence 12 and is T.

[0144] The probe 13 contains an SNP (single nucleotide polymorphism) site 13, the SNP site 13 being the 63rd nucleotide of sequence 13 and being G, and the SNP site 14 being the 64th nucleotide of sequence 13 and being C.

[0145] The probe 14 contains an SNP (single nucleotide polymorphism) site 15, which is the 100th nucleotide of sequence 14 and is G.

[0146] The probe 15 contains an SNP (single nucleotide polymorphism) site 16, which is the 69th nucleotide of sequence 15 and is nucleotide A.

[0147] The probe 16 contains an SNP (single nucleotide polymorphism) site 17, which is the 20th nucleotide of sequence 16 and is C; the SNP site 18 is the 151st nucleotide of sequence 16 and is G.

[0148] The probe 17 contains an SNP (single nucleotide polymorphism) site 19, which is the 81st nucleotide of sequence 17 and is T.

[0149] The probe 18 contains an SNP (single nucleotide polymorphism) site 20, which is the 90th nucleotide of sequence 18 and is nucleotide A.

[0150] The probe 19 contains an SNP (single nucleotide polymorphism) site 21, which is the 99th nucleotide of sequence 19 and is nucleotide A.

[0151] The probe 20 contains an SNP (single nucleotide polymorphism) site 22, which is the 92nd nucleotide of sequence 20 and is T.

[0152] The probe 21 contains an SNP (single nucleotide polymorphism) site 23, which is the 71st nucleotide of sequence 21 and is nucleotide A.

[0153] The probe 22 contains an SNP (single nucleotide polymorphism) site 24, which is the 100th nucleotide of sequence 22 and is T.

[0154] The probe 23 contains an SNP (single nucleotide polymorphism) site 25, which is the 100th nucleotide of sequence 23 and is T.

[0155] The probe 24 contains an SNP (single nucleotide polymorphism) site 26, which is the 58th nucleotide of sequence 24 and is T.

[0156] The probe 25 contains an SNP (single nucleotide polymorphism) site 27, which is the 100th nucleotide of sequence 25 and is A, and the SNP site 28 is the 136th nucleotide of sequence 25 and is T.

[0157] The probe 26 contains an SNP (single nucleotide polymorphism) site 29, which is the 78th nucleotide of sequence 26 and is T.

[0158] The probe 27 contains an SNP (single nucleotide polymorphism) site 30, which is the 100th nucleotide of sequence 27 and is T.

[0159] The probe 28 contains an SNP (single nucleotide polymorphism) site 31, which is the 69th nucleotide of sequence 28 and is T.

[0160] The nucleotide sequence of probe 1 is sequence 1 and the nucleotide of SNP site 1 is T;

[0161] The nucleotide sequence of probe 2 is sequence 2 and the nucleotide of SNP site 2 is T;

[0162] The nucleotide sequence of probe 3 is sequence 3 and the nucleotide of SNP site 3 is T;

[0163] The nucleotide sequence of probe 4 is sequence 4 and the nucleotide of SNP site 4 is A;

[0164] The nucleotide sequence of probe 5 is sequence 5 and the nucleotide of SNP site 5 is T;

[0165] The nucleotide sequence of probe 6 is sequence 6 and the nucleotide of SNP site 6 is C;

[0166] The nucleotide sequence of probe 7 is sequence 7 and the nucleotide of SNP site 7 is C;

[0167] The nucleotide sequence of probe 8 is sequence 8 and the nucleotide of SNP site 8 is C;

[0168] The nucleotide sequence of probe 9 is sequence 9 and the nucleotide of SNP site 9 is A;

[0169] The nucleotide sequence of probe 10 is sequence 10 and the nucleotide of SNP site 10 is A;

[0170] The nucleotide sequence of probe 11 is sequence 11 and the nucleotide of SNP site 11 is C;

[0171] The nucleotide sequence of probe 12 is sequence 12 and the nucleotide of SNP site 12 is T;

[0172] The nucleotide sequence of probe 13 is sequence 13, the nucleotide of SNP site 13 is G, and the nucleotide of SNP site 14 is C.

[0173] The nucleotide sequence of probe 14 is sequence 14 and the nucleotide of SNP site 15 is G;

[0174] The nucleotide sequence of probe 15 is sequence 15 and the nucleotide of SNP site 16 is A;

[0175] The nucleotide sequence of probe 16 is sequence 16, and the nucleotide of SNP site 17 is C; and the nucleotide of SNP site 18 is G;

[0176] The nucleotide sequence of probe 17 is sequence 17 and the nucleotide of SNP site 19 is T;

[0177] The nucleotide sequence of probe 18 is sequence 18 and the nucleotide of SNP site 20 is A;

[0178] The nucleotide sequence of probe 19 is sequence 19 and the nucleotide of SNP site 21 is A;

[0179] The nucleotide sequence of probe 20 is sequence 20 and the nucleotide of SNP site 22 is T;

[0180] The nucleotide sequence of probe 21 is sequence 21 and the nucleotide of SNP site 23 is A;

[0181] The nucleotide sequence of probe 22 is sequence 22 and the nucleotide of SNP site 24 is T;

[0182] The nucleotide sequence of probe 23 is sequence 23 and the nucleotide of SNP site 25 is T;

[0183] The nucleotide sequence of probe 24 is sequence 24 and the nucleotide of SNP site 26 is T;

[0184] The nucleotide sequence of probe 25 is sequence 25, the nucleotide of SNP site 27 is A, and the nucleotide of SNP site 28 is T.

[0185] The nucleotide sequence of probe 26 is sequence 26 and the nucleotide of SNP site 29 is T;

[0186] The nucleotide sequence of probe 27 is sequence 27 and the nucleotide of SNP site 30 is T;

[0187] The nucleotide sequence of probe 28 is sequence 28 and the nucleotide of SNP site 31 is T.

[0188] This invention also provides the application of the probe described above in any of the following:

[0189] (1) Identification or auxiliary identification of Brucella bovis;

[0190] (2) Screening or breeding of Brucella bovis;

[0191] (3) Prepare products for identification or auxiliary identification of Brucella bovis;

[0192] (4) Prepare products for screening or breeding of Brucella bovis.

[0193] This invention involves in-depth research on the whole genome sequences of Brucella bovis and non-Brucella bovis strains in the NCBI database. Using a local Blast database, a Python program was written to perform extensive screening and comparison. This identified conserved DNA regions in the Brucella bovis genome sequence and screened for specific probe sequences. These probes were then used to detect sequencing results (including second- or third-generation sequencing) of unknown samples. Since up to 28 specific probes are used, the presence of Brucella bovis can be determined as long as any one probe is detected. The more probes detected, the higher the reliability. This ensures that even genome sequences with poor sequencing quality and low coverage can be queried using these electron probes, enabling rapid, simple, and efficient identification of whether a sample is Brucella bovis. Attached Figure Description

[0194] Figure 1 This is a strategy for extracting specific tags.

[0195] Figure 2This shows the BLAST test results for the Babortus_Probe001 probe in the NCBI database. A represents strains with a completely identical specific sequence, and B represents strains with a different specific sequence.

[0196] Figure 3 Examples of specific BLAST results for Babortus_Probe001 in a single strain. A represents a specific genomic sequence that is completely identical to the specific sequence, and B represents a specific genomic sequence that is inconsistent with the specific sequence.

[0197] Figure 4 Examples of the practicality and specificity of specific probes.

[0198] Figure 5 This is an example of the application of specific probes using strain data from the literature. Detailed Implementation

[0199] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0200] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following examples are commercially available.

[0201] Unless otherwise specified, all quantitative experiments in the following examples are performed in triplicate.

[0202] The Brucella bovis genome sequence described in this article is the Babortus_001.fna chromosome sequence (GCA_000054005.1, GenBank:NC_007618.1, NC_007624.1), and the nucleic acid sequence positions are also calculated based on (GCA_000054005.1, GenBank:NC_007618.1, NC_007624.1).

[0203] Example 1: Acquisition of an electron probe for Brucella bovis

[0204] The specific steps for preparing the bovine Brucella electron probe are as follows:

[0205] 1. Download Brucella genome sequences from the NCBI Genome Database, totaling 800 strains of 12 species (Table 1), and number these sequences.

[0206] Table 1. Information on Brucella species and their numbers

[0207]

[0208]

[0209] 2. To ensure the reliability of the results, the quality of the downloaded genome sequences was analyzed using CheckM software, along with whole-genome SNP analysis and whole-genome core gene clustering analysis. Strains with obvious classification errors were removed, leaving 789 strains from 8 species for analysis (Table 2).

[0210] Table 2 Information on the remaining strains selected for analysis

[0211]

[0212] 3. Using the chromosome sequence of Babortus_001.fna (GCA_000054005.1, (NC_007618.1, NC_007624.1)) as the reference sequence, sequence tags were extracted according to window=100bp and step=1bp, resulting in 3,278,208 100bp sequence tags. The results are as follows. Figure 1 As shown.

[0213] 4. Using a local BLAST (BLAST-2.7.1+), a database was built using the genome sequences of 533 Brucella strains (excluding *Brucella bovis*) as the query sequences, with 3,278,208 sequence tags as the query sequences. A Python program was used to remove all the retrieved sequences (Identical = 100%, alignment length = 100 bp). This step identified 3,006,001 sequence tags in the genome sequences of the other 533 Brucella strains; after removing these tags, 272,207 sequence tags remained.

[0214] 5. Locate sequence tags present in all 256 strains of Brucella bovis.

[0215] A Python program was written to query the genome sequences of all 256 Brucella bovis strains using the 272,207 sequence tags obtained in "Step 4". Sequence tags that were not present in the genome of any strain were removed. As a result, we obtained 25,960 sequence tags that were present in the genomes of all 256 Brucella bovis strains.

[0216] 6. Based on the overlap between fragments, 25,960 sequence tags were fused to obtain 488 specific genomic fragments, which ranged in length from 100bp to 321bp.

[0217] 7. Using these 488 fragments, perform a local BLAST query on all Brucella genomes. Based on the "subject start" and "subject end" information in the BLAST results, extract the information of each fragment in each genome. Construct a FASTA file from the fragments that match the genomes of each of these 488 fragments, resulting in a total of 488 FASTA files.

[0218] 8. The 488 FASTA format files obtained in step 7 were compared using MegaX software to find specific SNP sites that could distinguish Brucella abortus: the candidate sites were the same bases in all Brucella abortus strains, while in all Non-B. abortus strains they were different bases from B. abortus.

[0219] 9. A total of 385 sequences contained one (or more) SNP sites. For ease of use, all sequences containing two or more SNP sites were selected. For sequences containing only one SNP site, the SNP site must be more than 50 bp away from both ends of the sequence. A total of 28 electron probes were obtained (Table 3).

[0220] Table 3. Specific electron probe sequences and SNP sites (underlined bases)

[0221]

[0222]

[0223]

[0224]

[0225] Example 2: Specificity verification of 28 electron probes

[0226] To test the specificity of these 28 electron probes in a larger database, an online BLAST query was performed on them (https: / / blast.ncbi.nlm.nih.gov / Blast.cgi). The results showed that all sequences with QueryCoverage=100% and percent identity=100% were B. abortus genome sequences, indicating that these 28 sequences have very good specificity.

[0227] The test results using Babortus_Probe01 as an example are as follows: Figure 2 ,in Figure 2 In the sequence A, it represents a genome sequence that is completely identical to the specific sequence. Figure 2 The letter B indicates a genomic sequence inconsistent with the specific sequence. Results show that this electronic probe achieved 100% Query Cover and Per.Ident (Percentage of Identity) values ​​only in *B. abortus*. See the attached diagram for details. Figure 3 The sequence of A is completely identical; it cannot be found in any strain of No-B. abortus, as shown in the following figure. Figure 3 In case B, the Query Cover and Per.Ident values ​​cannot both be 100%, indicating that the sequences are not completely consistent.

[0228] Genome acquisition of test strains: 1240 Brucella genomes were downloaded from the website (https: / / www.bv-brc.org / view / Taxonomy / 234#view_tab=genomes). 758 of these strains overlapped with the 800 strains previously used, resulting in 482 newly added genomes. The sequencing quality of these 482 strains was assessed using CheckM, and whole-genome SNP analysis was performed. Strains with obvious classification errors (8 strains) were removed, leaving 474 strains. Strains classified only to the genus level (10 strains) were then removed, leaving 464 usable strains. Information is shown in Table 4 below.

[0229] Table 4. Information on Brucella strains used in the case tests.

[0230]

[0231]

[0232] The 464 strains in Table 4 and 10271 strains of bacteria from genera (species) such as Bacillus, Acinetobacter, and Yersinia downloaded from the Internet (https: / / www.ncbi.nlm.nih.gov / ) were identified and analyzed using 28 specific probes for identifying Brucella bovis.

[0233] Table 5 Summary of other strains involved in the case study

[0234]

[0235] All the above-mentioned strains were used as test strains. The genome sequencing results of the test strains were compared with the specific probes for identifying Brucella bovis. The sequence alignment results were used to determine whether the strain was Brucella bovis.

[0236] If the sequence alignment results indicate that the genomic DNA sequence of the test strain contains any one of the 28 specific probes, the test strain is Brucella bovis or contains Brucella bovis; if the sequence alignment results indicate that the genomic DNA sequence of the test sample does not contain any one of the 28 specific probes, the test sample is not Brucella bovis or does not contain Brucella bovis.

[0237] The 28 specific probes contain at least one of 28 single-stranded DNAs named probe 1 through probe 28:

[0238] Probe 1 contains SNP (single nucleotide polymorphism) site 1, which is the 100th nucleotide of sequence 1 and is T; where y in sequence 1 in the sequence listing represents t or c.

[0239] Probe 2 contains SNP (single nucleotide polymorphism) site 2, which is the 100th nucleotide of sequence 2 and is T; where y in sequence 2 in the sequence listing represents t or c.

[0240] Probe 3 contains SNP (single nucleotide polymorphism) site 3, which is the 100th nucleotide of sequence 3 and is T; where y in sequence 3 in the sequence listing represents t or c.

[0241] Probe 4 contains SNP (single nucleotide polymorphism) site 4, which is the 100th nucleotide of sequence 4 and is A; where r in sequence 4 in the sequence listing represents a or g.

[0242] Probe 5 contains an SNP (single nucleotide polymorphism) site 5, which is the 100th nucleotide of sequence 5 and is T; where w in sequence 5 in the sequence listing represents t or a.

[0243] Probe 6 contains an SNP (single nucleotide polymorphism) site 6, which is the 92nd nucleotide of sequence 6 and is C; where y in sequence 6 in the sequence listing represents c or t.

[0244] Probe 7 contains an SNP (single nucleotide polymorphism) site 7, which is the 63rd nucleotide of sequence 7 and is C; where 's' in sequence 7 in the sequence listing represents c or g.

[0245] Probe 8 contains SNP (single nucleotide polymorphism) site 8, which is the 100th nucleotide of sequence 8 and is C; where y in sequence 8 in the sequence listing represents c or t.

[0246] Probe 9 contains an SNP (single nucleotide polymorphism) site 9, which is the 68th nucleotide of sequence 9 and is A; where r in sequence 9 in the sequence listing represents a or g.

[0247] Probe 10 contains an SNP (single nucleotide polymorphism) site 10, which is the 79th nucleotide of sequence 10 and is A; wherein m in sequence 10 in the sequence listing represents a or c.

[0248] Probe 11 contains an SNP (single nucleotide polymorphism) site 11, which is the 74th nucleotide of sequence 11 and is C; wherein y in sequence 11 in the sequence listing represents c or t.

[0249] Probe 12 contains an SNP (single nucleotide polymorphism) site 12, which is the 87th nucleotide of sequence 12 and is T; wherein y in sequence 12 in the sequence listing represents c or t.

[0250] Probe 13 contains an SNP (single nucleotide polymorphism) site 13, wherein the SNP site 13 is the 63rd nucleotide of sequence 13 and the nucleotide is G, and the SNP site 14 is the 64th nucleotide of sequence 13 and the nucleotide is C; wherein in the sequence listing, r represents g or a, and m represents c or a.

[0251] Probe 14 contains an SNP (single nucleotide polymorphism) site 15, which is the 100th nucleotide of sequence 14 and is G; where r in sequence 14 in the sequence listing represents g or a.

[0252] Probe 15 contains an SNP (single nucleotide polymorphism) site 16, which is the 69th nucleotide of sequence 15 and is A; where r in sequence 15 in the sequence listing represents g or a.

[0253] Probe 16 contains an SNP (single nucleotide polymorphism) site 17, which is the 20th nucleotide of sequence 16 and is C; and an SNP site 18, which is the 151st nucleotide of sequence 16 and is G; wherein r in sequence 16 in the sequence listing represents g or a, and y represents c or t.

[0254] Probe 17 contains an SNP (single nucleotide polymorphism) site 19, which is the 81st nucleotide of sequence 17 and is T; where y in sequence 17 in the sequence listing represents c or t.

[0255] Probe 18 contains an SNP (single nucleotide polymorphism) site 20, which is the 90th nucleotide of sequence 18 and is A; where r in sequence 18 in the sequence listing represents g or a.

[0256] Probe 19 contains an SNP (single nucleotide polymorphism) site 21, which is the 99th nucleotide of sequence 19 and is A; where w in sequence 19 in the sequence listing represents t or a.

[0257] Probe 20 contains an SNP (single nucleotide polymorphism) site 22, which is the 92nd nucleotide of sequence 20 and is T; wherein y in sequence 20 in the sequence listing represents c or t.

[0258] Probe 21 contains an SNP (single nucleotide polymorphism) site 23, which is the 71st nucleotide of sequence 21 and is A; where m in sequence 21 in the sequence listing represents c or a.

[0259] Probe 22 contains an SNP (single nucleotide polymorphism) site 24, which is the 100th nucleotide of sequence 22 and is T; where y in sequence 22 in the sequence listing represents c or t.

[0260] Probe 23 contains an SNP (single nucleotide polymorphism) site 25, which is the 100th nucleotide of sequence 23 and is T; wherein y in sequence 23 in the sequence listing represents c or t.

[0261] Probe 24 contains an SNP (single nucleotide polymorphism) site 26, which is the 58th nucleotide of sequence 24 and is T; where y in sequence 24 in the sequence listing represents c or t.

[0262] Probe 25 contains an SNP (single nucleotide polymorphism) site 27, which is the 100th nucleotide of sequence 25 and is A, and an SNP site 28, which is the 136th nucleotide of sequence 25 and is T; wherein y in sequence 25 in the sequence listing represents c or t, and m represents a or c.

[0263] Probe 26 contains an SNP (single nucleotide polymorphism) site 29, which is the 78th nucleotide of sequence 26 and is T; where y in sequence 26 in the sequence listing represents c or t.

[0264] Probe 27 contains an SNP (single nucleotide polymorphism) site 30, which is the 100th nucleotide of sequence 27 and is T; where y in sequence 27 in the sequence listing represents c or t.

[0265] Probe 28 contains an SNP (single nucleotide polymorphism) site 31, which is the 69th nucleotide of sequence 28 and is T; where y in sequence 28 in the sequence listing represents c or t.

[0266] The results showed that all 223 *Brucella abortus* strains contained the aforementioned 28 specific probes, while the genomic DNA sequences of other strains did not contain any of these 28 specific probes. Figure 4 This application example further demonstrates the specificity and practicality of these probes.

[0267] Example 3: Application of 28 electron probes in the identification of Brucella bovis

[0268] A review of the literature [Pathogens, 2021, 10(6), 759; DOI: 10.3390 / pathogens10060759] revealed that the authors isolated 29 species of Brucella from the lymph nodes, milk, and abomasal contents of infected cattle, buffalo, sheep, and goats in nine regions of Egypt. The isolates were identified using several methods, including microbiological methods, matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF MS), and multiplex PCR. Finally, Illumina was used again to analyze the isolates. Sequencing of 29 Brucella isolates and phylogenetic clustering revealed 8 species of B. abortus and 21 species of B. melitensis.

[0269] Twenty-nine Brucella species were obtained from the website (https: / / www.ncbi.nlm.nih.gov / biosample?LinkName=bioproject_biosample_all&from_uid=650270), and specific strain information is shown in Table 6. After simple sequence assembly of these SRA files, they were compared using the 28 electron probes described above. The results showed that all eight Brucella abortus strains contained the 28 specific probes described in Example 2 (such as...). Figure 5 The upper half of the black square shows that the genomic DNA of the other 21 Brucella melitensis strains did not contain any of the 28 specific probes described in Example 2 (such as...). Figure 5 (The lower half is shown in gray squares). The *Brucella bovis* strains identified using the 28 probes of this invention were completely consistent with the results in the literature, demonstrating speed and accuracy. This application example further proves the specificity and practicality of these probes.

[0270] Table 6. Information on strains from the literature and identification results using the 28 probes of this invention.

[0271] SRA number BioSample Number Document number Authentication results of the documents Identification results using the probe of this invention SRR12368029 SAMN15701920 18RB17233 Brucella abortus Brucella abortus SRR12368032 SAMN15701943 18RB17259 Brucella abortus Brucella abortus SRR12368034 SAMN15701941 18RB17257 Brucella abortus Brucella abortus SRR12368035 SAMN15701940 18RB17256 Brucella abortus Brucella abortus SRR12368036 SAMN15701939 18RB17255 Brucella abortus Brucella abortus SRR12368047 SAMN15701929 18RB17245 Brucella abortus Brucella abortus SRR12368049 SAMN15701927 18RB17243 Brucella abortus Brucella abortus SRR12368050 SAMN15701926 18RB17242 Brucella abortus Brucella abortus SRR12368024 SAMN15701925 18RB17241 Brucella melitensis non-Brucella abortus SRR12368025 SAMN15701924 18RB17240 Brucella melitensis non-Brucella abortus SRR12368026 SAMN15701923 18RB17238 Brucella melitensis non-Brucella abortus SRR12368027 SAMN15701922 18RB17236 Brucella melitensis non-Brucella abortus SRR12368028 SAMN15701921 18RB17235 Brucella melitensis non-Brucella abortus SRR12368030 SAMN15701919 18RB17230 Brucella melitensis non-Brucella abortion SRR12368031 SAMN15701944 18RB17260 Brucella melitensis non-Brucella abortion SRR12368033 SAMN15701942 18RB17258 Brucella melitensis non-Brucella abortion SRR12368037 SAMN15701938 18RB17254 Brucella melitensis non-Brucella abortion SRR12368038 SAMN15701937 18RB17253 Brucella melitensis non-Brucella abortion SRR12368039 SAMN15701936 18RB17252 Brucella melitensis non-Brucella abortion SRR12368040 SAMN15701918 18RB17229 Brucella melitensis non-Brucella abortion SRR12368041 SAMN15701935 18RB17251 Brucella melitensis non-Brucella abortion SRR12368042 SAMN15701934 18RB17250 Brucella melitensis non-Brucella abortion SRR12368043 SAMN15701933 18RB17249 Brucella melitensis non-Brucella abortion SRR12368044 SAMN15701932 18RB17248 Brucella melitensis non-Brucella abortion SRR12368045 SAMN15701931 18RB17247 Brucella melitensis non-Brucella abortion SRR12368046 SAMN15701930 18RB17246 Brucella melitensis non-Brucella abortion SRR12368048 SAMN15701928 18RB17244 Brucella melitensis non-Brucella abortion SRR12368051 SAMN15701917 18RB17228 Brucella melitensis non-Brucella abortion SRR12368052 SAMN15701916 18RB17227 Brucella melitensis non-Brucella abortion

[0272] The specific steps for identifying Brucella bovis using 28 electron probes are as follows:

[0273] 1. Collect the bacteria to be tested and extract their genome. The whole genome extraction of bacteria was performed according to the procedure of the Bacterial Genomic DNA Extraction kit (Promega, catalog number A1120);

[0274] 2. Genome samples were sequenced using the Nanopore / Illumina platform;

[0275] 3. Use bioinformatics software to compare and analyze the sequencing results with the probe sequences:

[0276] The sequencing results were compared with the 28 specific probes described in Example 2 for identifying Brucella bovis. The sequence alignment results were used to determine, or to assist in determining, whether the test bacterium was Brucella bovis: if the sequence alignment results showed that the genomic DNA sequence of the test bacterium contained any one of the 28 specific probes described in Example 2, the test bacterium was Brucella bovis; if the sequence alignment results showed that the genomic DNA sequence of the test bacterium did not contain any of the 28 specific probes described in Example 2, the test bacterium was not Brucella bovis. Multiple electron probes can avoid incorrect identification due to poor sequencing quality or artificially modified bacteria; the more probes detected, the more reliable the results.

[0277] The present invention has been described in detail above. For those skilled in the art, the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope, and without requiring unnecessary experiments. Although specific embodiments have been given, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including changes made using conventional techniques known in the art that depart from the scope disclosed herein. Some of the essential features can be applied within the scope of the following appended claims.

Claims

1. A method for identifying Brucella bovis for non-disease diagnostic and therapeutic purposes, characterized in that: The method includes the following steps: S1) Extract genomic DNA from the sample to be tested; S2) Sequencing the genomic DNA to obtain sequencing results; S3) The sequencing results are compared with the specific probe for identifying Brucella bovis. Based on the sequence comparison results, it is determined or assisted in determining whether the sample to be tested is Brucella bovis or whether it contains Brucella bovis. If the sequence alignment results show that the genomic DNA sequence of the sample to be tested contains any of the specific probes, the sample to be tested is Brucella bovis or contains Brucella bovis; if the sequence alignment results show that the genomic DNA sequence of the sample to be tested does not contain the specific probes, the sample to be tested is not Brucella bovis or does not contain Brucella bovis. The specific probes comprise 28 single-stranded DNAs named probe 1 through probe 28: Probe 1 contains SNP (single nucleotide polymorphism) site 1, which is the 100th nucleotide of sequence 1 and is T; Probe 2 contains SNP (single nucleotide polymorphism) site 2, which is the 100th nucleotide of sequence 2 and is T; Probe 3 contains SNP (single nucleotide polymorphism) site 3, which is the 100th nucleotide of sequence 3 and is T; Probe 4 contains SNP (single nucleotide polymorphism) site 4, which is the 100th nucleotide of sequence 4 and is A; Probe 5 contains SNP (single nucleotide polymorphism) site 5, which is the 100th nucleotide of sequence 5 and is T; Probe 6 contains SNP (single nucleotide polymorphism) site 6, which is the 92nd nucleotide of sequence 6 and is C; Probe 7 contains SNP (single nucleotide polymorphism) site 7, which is the 63rd nucleotide of sequence 7 and is C. Probe 8 contains SNP (single nucleotide polymorphism) site 8, which is the 100th nucleotide of sequence 8 and is C. Probe 9 contains SNP (single nucleotide polymorphism) site 9, which is the 68th nucleotide of sequence 9 and is A; Probe 10 contains an SNP (single nucleotide polymorphism) site 10, which is the 79th nucleotide of sequence 10 and is A. Probe 11 contains an SNP (single nucleotide polymorphism) site 11, which is the 74th nucleotide of sequence 11 and is C. Probe 12 contains an SNP (single nucleotide polymorphism) site 12, which is the 87th nucleotide of sequence 12 and is T. Probe 13 contains SNP (single nucleotide polymorphism) sites 13 and 14, wherein SNP site 13 is the 63rd nucleotide of sequence 13 and is G, and SNP site 14 is the 64th nucleotide of sequence 13 and is C. Probe 14 contains an SNP (single nucleotide polymorphism) site 15, which is the 100th nucleotide of sequence 14 and is G; Probe 15 contains an SNP (single nucleotide polymorphism) site 16, which is the 69th nucleotide of sequence 15 and is nucleotide A; Probe 16 contains SNP (single nucleotide polymorphism) sites 17 and 18, wherein SNP site 17 is the 20th nucleotide of sequence 16 and is C; and SNP site 18 is the 151st nucleotide of sequence 16 and is G. Probe 17 contains an SNP (single nucleotide polymorphism) site 19, which is the 81st nucleotide of sequence 17 and is T. Probe 18 contains an SNP (single nucleotide polymorphism) site 20, which is the 90th nucleotide of sequence 18 and is nucleotide A; Probe 19 contains an SNP (single nucleotide polymorphism) site 21, which is the 99th nucleotide of sequence 19 and is nucleotide A; Probe 20 contains an SNP (single nucleotide polymorphism) site 22, which is the 92nd nucleotide of sequence 20 and is T. Probe 21 contains an SNP (single nucleotide polymorphism) site 23, which is the 71st nucleotide of sequence 21 and is nucleotide A; Probe 22 contains an SNP (single nucleotide polymorphism) site 24, which is the 100th nucleotide of sequence 22 and is T. Probe 23 contains an SNP (single nucleotide polymorphism) site 25, which is the 100th nucleotide of sequence 23 and is T. Probe 24 contains an SNP (single nucleotide polymorphism) site 26, which is the 58th nucleotide of sequence 24 and is T. Probe 25 contains SNP (single nucleotide polymorphism) sites 27 and 28, wherein SNP site 27 is the 100th nucleotide of sequence 25 and is A, and SNP site 28 is the 136th nucleotide of sequence 25 and is T. Probe 26 contains an SNP (single nucleotide polymorphism) site 29, which is the 78th nucleotide of sequence 26 and is T. Probe 27 contains an SNP (single nucleotide polymorphism) site 30, which is the 100th nucleotide of sequence 27 and is T. Probe 28 contains an SNP (single nucleotide polymorphism) site 31, which is the 69th nucleotide of sequence 28 and is T.

2. The method according to claim 1, characterized in that: The nucleotide sequence of probe 1 is sequence 1 and the nucleotide of SNP site 1 is T; The nucleotide sequence of probe 2 is sequence 2 and the nucleotide of SNP site 2 is T; The nucleotide sequence of probe 3 is sequence 3 and the nucleotide of SNP site 3 is T; The nucleotide sequence of probe 4 is sequence 4 and the nucleotide of SNP site 4 is A; The nucleotide sequence of probe 5 is sequence 5 and the nucleotide of SNP site 5 is T; The nucleotide sequence of probe 6 is sequence 6 and the nucleotide of SNP site 6 is C; The nucleotide sequence of probe 7 is sequence 7 and the nucleotide of SNP site 7 is C; The nucleotide sequence of probe 8 is sequence 8 and the nucleotide of SNP site 8 is C; The nucleotide sequence of probe 9 is sequence 9 and the nucleotide of SNP site 9 is A; The nucleotide sequence of probe 10 is sequence 10 and the nucleotide of SNP site 10 is A; The nucleotide sequence of probe 11 is sequence 11 and the nucleotide of SNP site 11 is C; The nucleotide sequence of probe 12 is sequence 12 and the nucleotide of SNP site 12 is T; The nucleotide sequence of probe 13 is sequence 13, the nucleotide of SNP site 13 is G, and the nucleotide of SNP site 14 is C. The nucleotide sequence of probe 14 is sequence 14 and the nucleotide of SNP site 15 is G; The nucleotide sequence of probe 15 is sequence 15 and the nucleotide of SNP site 16 is A; The nucleotide sequence of probe 16 is sequence 16, and the nucleotide of SNP site 17 is C; and the nucleotide of SNP site 18 is G; The nucleotide sequence of probe 17 is sequence 17 and the nucleotide of SNP site 19 is T; The nucleotide sequence of probe 18 is sequence 18 and the nucleotide of SNP site 20 is A; The nucleotide sequence of probe 19 is sequence 19 and the nucleotide of SNP site 21 is A; The nucleotide sequence of probe 20 is sequence 20 and the nucleotide of SNP site 22 is T; The nucleotide sequence of probe 21 is sequence 21 and the nucleotide of SNP site 23 is A; The nucleotide sequence of probe 22 is sequence 22 and the nucleotide of SNP site 24 is T; The nucleotide sequence of probe 23 is sequence 23 and the nucleotide of SNP site 25 is T; The nucleotide sequence of probe 24 is sequence 24 and the nucleotide of SNP site 26 is T; The nucleotide sequence of probe 25 is sequence 25, the nucleotide of SNP site 27 is A, and the nucleotide of SNP site 28 is T. The nucleotide sequence of probe 26 is sequence 26 and the nucleotide of SNP site 29 is T; The nucleotide sequence of probe 27 is sequence 27 and the nucleotide of SNP site 30 is T; The nucleotide sequence of probe 28 is sequence 28 and the nucleotide of SNP site 31 is T.

3. An apparatus for identifying Brucella bovis, the apparatus comprising: 1) Sequence Alignment Module: This module aligns the genomic DNA of the sample to be tested with specific probes for identifying Brucella bovis and outputs the alignment results. The specific probes comprise 28 single-stranded DNA molecules named probe 1 to probe 28. Probe 1 contains SNP (single nucleotide polymorphism) site 1, which is the 100th nucleotide of sequence 1 and is T; Probe 2 contains SNP (single nucleotide polymorphism) site 2, which is the 100th nucleotide of sequence 2 and is T; Probe 3 contains SNP (single nucleotide polymorphism) site 3, which is the 100th nucleotide of sequence 3 and is T; Probe 4 contains SNP (single nucleotide polymorphism) site 4, which is the 100th nucleotide of sequence 4 and is A; Probe 5 contains SNP (single nucleotide polymorphism) site 5, which is the 100th nucleotide of sequence 5 and is T; Probe 6 contains SNP (single nucleotide polymorphism) site 6, which is the 92nd nucleotide of sequence 6 and is C; Probe 7 contains SNP (single nucleotide polymorphism) site 7, which is the 63rd nucleotide of sequence 7 and is C. Probe 8 contains SNP (single nucleotide polymorphism) site 8, which is the 100th nucleotide of sequence 8 and is C. Probe 9 contains SNP (single nucleotide polymorphism) site 9, which is the 68th nucleotide of sequence 9 and is A; Probe 10 contains an SNP (single nucleotide polymorphism) site 10, which is the 79th nucleotide of sequence 10 and is A. Probe 11 contains an SNP (single nucleotide polymorphism) site 11, which is the 74th nucleotide of sequence 11 and is C. Probe 12 contains an SNP (single nucleotide polymorphism) site 12, which is the 87th nucleotide of sequence 12 and is T. Probe 13 contains SNP (single nucleotide polymorphism) sites 13 and 14, wherein SNP site 13 is the 63rd nucleotide of sequence 13 and is G, and SNP site 14 is the 64th nucleotide of sequence 13 and is C. Probe 14 contains an SNP (single nucleotide polymorphism) site 15, which is the 100th nucleotide of sequence 14 and is G; Probe 15 contains an SNP (single nucleotide polymorphism) site 16, which is the 69th nucleotide of sequence 15 and is nucleotide A; Probe 16 contains SNP (single nucleotide polymorphism) sites 17 and 18, wherein SNP site 17 is the 20th nucleotide of sequence 16 and is C; and SNP site 18 is the 151st nucleotide of sequence 16 and is G. Probe 17 contains an SNP (single nucleotide polymorphism) site 19, which is the 81st nucleotide of sequence 17 and is T. Probe 18 contains an SNP (single nucleotide polymorphism) site 20, which is the 90th nucleotide of sequence 18 and is nucleotide A; Probe 19 contains an SNP (single nucleotide polymorphism) site 21, which is the 99th nucleotide of sequence 19 and is nucleotide A; Probe 20 contains an SNP (single nucleotide polymorphism) site 22, which is the 92nd nucleotide of sequence 20 and is T. Probe 21 contains an SNP (single nucleotide polymorphism) site 23, which is the 71st nucleotide of sequence 21 and is nucleotide A; Probe 22 contains an SNP (single nucleotide polymorphism) site 24, which is the 100th nucleotide of sequence 22 and is T. Probe 23 contains an SNP (single nucleotide polymorphism) site 25, which is the 100th nucleotide of sequence 23 and is T. Probe 24 contains an SNP (single nucleotide polymorphism) site 26, which is the 58th nucleotide of sequence 24 and is T. Probe 25 contains SNP (single nucleotide polymorphism) sites 27 and 28, wherein SNP site 27 is the 100th nucleotide of sequence 25 and is A, and SNP site 28 is the 136th nucleotide of sequence 25 and is T. Probe 26 contains an SNP (single nucleotide polymorphism) site 29, which is the 78th nucleotide of sequence 26 and is T. Probe 27 contains an SNP (single nucleotide polymorphism) site 30, which is the 100th nucleotide of sequence 27 and is T. Probe 28 contains an SNP (single nucleotide polymorphism) site 31, which is the 69th nucleotide of sequence 28 and is T. 2) Identification module: Based on the sequence alignment results, determine or assist in determining whether the sample to be tested is Brucella bovis or whether it contains Brucella bovis; if the sequence alignment results show that the genomic DNA sequence of the sample to be tested contains any one of the specific probes, the sample to be tested is Brucella bovis or contains Brucella bovis; if the sequence alignment results show that the genomic DNA sequence of the sample to be tested does not contain the specific probe, the sample to be tested is not Brucella bovis or does not contain Brucella bovis.

4. The apparatus according to claim 3, characterized in that: The nucleotide sequence of probe 1 is sequence 1, and the nucleotide of SNP site 1 is T. The nucleotide sequence of probe 2 is sequence 2 and the nucleotide of SNP site 2 is T; The nucleotide sequence of probe 3 is sequence 3 and the nucleotide of SNP site 3 is T; The nucleotide sequence of probe 4 is sequence 4 and the nucleotide of SNP site 4 is A; The nucleotide sequence of probe 5 is sequence 5 and the nucleotide of SNP site 5 is T; The nucleotide sequence of probe 6 is sequence 6 and the nucleotide of SNP site 6 is C; The nucleotide sequence of probe 7 is sequence 7 and the nucleotide of SNP site 7 is C; The nucleotide sequence of probe 8 is sequence 8 and the nucleotide of SNP site 8 is C; The nucleotide sequence of probe 9 is sequence 9 and the nucleotide of SNP site 9 is A; The nucleotide sequence of probe 10 is sequence 10 and the nucleotide of SNP site 10 is A; The nucleotide sequence of probe 11 is sequence 11 and the nucleotide of SNP site 11 is C; The nucleotide sequence of probe 12 is sequence 12 and the nucleotide of SNP site 12 is T; The nucleotide sequence of probe 13 is sequence 13, the nucleotide of SNP site 13 is G, and the nucleotide of SNP site 14 is C. The nucleotide sequence of probe 14 is sequence 14 and the nucleotide of SNP site 15 is G; The nucleotide sequence of probe 15 is sequence 15 and the nucleotide of SNP site 16 is A; The nucleotide sequence of probe 16 is sequence 16, and the nucleotide of SNP site 17 is C; and the nucleotide of SNP site 18 is G; The nucleotide sequence of probe 17 is sequence 17 and the nucleotide of SNP site 19 is T; The nucleotide sequence of probe 18 is sequence 18 and the nucleotide of SNP site 20 is A; The nucleotide sequence of probe 19 is sequence 19 and the nucleotide of SNP site 21 is A; The nucleotide sequence of probe 20 is sequence 20 and the nucleotide of SNP site 22 is T; The nucleotide sequence of probe 21 is sequence 21 and the nucleotide of SNP site 23 is A; The nucleotide sequence of probe 22 is sequence 22 and the nucleotide of SNP site 24 is T; The nucleotide sequence of probe 23 is sequence 23 and the nucleotide of SNP site 25 is T; The nucleotide sequence of probe 24 is sequence 24 and the nucleotide of SNP site 26 is T; The nucleotide sequence of probe 25 is sequence 25, the nucleotide of SNP site 27 is A, and the nucleotide of SNP site 28 is T. The nucleotide sequence of probe 26 is sequence 26 and the nucleotide of SNP site 29 is T; The nucleotide sequence of probe 27 is sequence 27 and the nucleotide of SNP site 30 is T; The nucleotide sequence of probe 28 is sequence 28 and the nucleotide of SNP site 31 is T.

5. A computer-readable storage medium storing a computer program, characterized in that: The computer program causes the computer to perform the steps of the method as described in claim 1 or 2.

6. The use of the apparatus according to claim 3 or 4 in the preparation of products for identifying Brucella bovis.

7. The use of the computer-readable storage medium of claim 5 in the preparation of products for identifying Brucella bovis.

8. A specific probe for identifying Brucella bovis, characterized in that: The specific probes comprise 28 single-stranded DNAs named probe 1 to probe 28. Probe 1 contains SNP (single nucleotide polymorphism) site 1, which is the 100th nucleotide of sequence 1 and is T; Probe 2 contains SNP (single nucleotide polymorphism) site 2, which is the 100th nucleotide of sequence 2 and is T; Probe 3 contains SNP (single nucleotide polymorphism) site 3, which is the 100th nucleotide of sequence 3 and is T; Probe 4 contains SNP (single nucleotide polymorphism) site 4, which is the 100th nucleotide of sequence 4 and is A; Probe 5 contains SNP (single nucleotide polymorphism) site 5, which is the 100th nucleotide of sequence 5 and is T; Probe 6 contains SNP (single nucleotide polymorphism) site 6, which is the 92nd nucleotide of sequence 6 and is C; Probe 7 contains SNP (single nucleotide polymorphism) site 7, which is the 63rd nucleotide of sequence 7 and is C. Probe 8 contains SNP (single nucleotide polymorphism) site 8, which is the 100th nucleotide of sequence 8 and is C. Probe 9 contains SNP (single nucleotide polymorphism) site 9, which is the 68th nucleotide of sequence 9 and is A; Probe 10 contains an SNP (single nucleotide polymorphism) site 10, which is the 79th nucleotide of sequence 10 and is A. Probe 11 contains an SNP (single nucleotide polymorphism) site 11, which is the 74th nucleotide of sequence 11 and is C. Probe 12 contains an SNP (single nucleotide polymorphism) site 12, which is the 87th nucleotide of sequence 12 and is T. Probe 13 contains SNP (single nucleotide polymorphism) sites 13 and 14, wherein SNP site 13 is the 63rd nucleotide of sequence 13 and is G, and SNP site 14 is the 64th nucleotide of sequence 13 and is C. Probe 14 contains an SNP (single nucleotide polymorphism) site 15, which is the 100th nucleotide of sequence 14 and is G; Probe 15 contains an SNP (single nucleotide polymorphism) site 16, which is the 69th nucleotide of sequence 15 and is nucleotide A; Probe 16 contains SNP (single nucleotide polymorphism) sites 17 and 18, wherein SNP site 17 is the 20th nucleotide of sequence 16 and is C; and SNP site 18 is the 151st nucleotide of sequence 16 and is G. Probe 17 contains an SNP (single nucleotide polymorphism) site 19, which is the 81st nucleotide of sequence 17 and is T. Probe 18 contains an SNP (single nucleotide polymorphism) site 20, which is the 90th nucleotide of sequence 18 and is nucleotide A; Probe 19 contains an SNP (single nucleotide polymorphism) site 21, which is the 99th nucleotide of sequence 19 and is nucleotide A; Probe 20 contains an SNP (single nucleotide polymorphism) site 22, which is the 92nd nucleotide of sequence 20 and is T. Probe 21 contains an SNP (single nucleotide polymorphism) site 23, which is the 71st nucleotide of sequence 21 and is nucleotide A; Probe 22 contains an SNP (single nucleotide polymorphism) site 24, which is the 100th nucleotide of sequence 22 and is T. Probe 23 contains an SNP (single nucleotide polymorphism) site 25, which is the 100th nucleotide of sequence 23 and is T. Probe 24 contains an SNP (single nucleotide polymorphism) site 26, which is the 58th nucleotide of sequence 24 and is T. Probe 25 contains SNP (single nucleotide polymorphism) sites 27 and 28, wherein SNP site 27 is the 100th nucleotide of sequence 25 and is A, and SNP site 28 is the 136th nucleotide of sequence 25 and is T. Probe 26 contains an SNP (single nucleotide polymorphism) site 29, which is the 78th nucleotide of sequence 26 and is T. Probe 27 contains an SNP (single nucleotide polymorphism) site 30, which is the 100th nucleotide of sequence 27 and is T. Probe 28 contains an SNP (single nucleotide polymorphism) site 31, which is the 69th nucleotide of sequence 28 and is T.

9. The specific probe according to claim 8, characterized in that: The nucleotide sequence of probe 1 is sequence 1, and the nucleotide of SNP site 1 is T. The nucleotide sequence of probe 2 is sequence 2 and the nucleotide of SNP site 2 is T; The nucleotide sequence of probe 3 is sequence 3 and the nucleotide of SNP site 3 is T; The nucleotide sequence of probe 4 is sequence 4 and the nucleotide of SNP site 4 is A; The nucleotide sequence of probe 5 is sequence 5 and the nucleotide of SNP site 5 is T; The nucleotide sequence of probe 6 is sequence 6 and the nucleotide of SNP site 6 is C; The nucleotide sequence of probe 7 is sequence 7 and the nucleotide of SNP site 7 is C; The nucleotide sequence of probe 8 is sequence 8 and the nucleotide of SNP site 8 is C; The nucleotide sequence of probe 9 is sequence 9 and the nucleotide of SNP site 9 is A; The nucleotide sequence of probe 10 is sequence 10 and the nucleotide of SNP site 10 is A; The nucleotide sequence of probe 11 is sequence 11 and the nucleotide of SNP site 11 is C; The nucleotide sequence of probe 12 is sequence 12 and the nucleotide of SNP site 12 is T; The nucleotide sequence of probe 13 is sequence 13, the nucleotide of SNP site 13 is G, and the nucleotide of SNP site 14 is C. The nucleotide sequence of probe 14 is sequence 14 and the nucleotide of SNP site 15 is G; The nucleotide sequence of probe 15 is sequence 15 and the nucleotide of SNP site 16 is A; The nucleotide sequence of probe 16 is sequence 16, and the nucleotide of SNP site 17 is C; and the nucleotide of SNP site 18 is G; The nucleotide sequence of probe 17 is sequence 17 and the nucleotide of SNP site 19 is T; The nucleotide sequence of probe 18 is sequence 18 and the nucleotide of SNP site 20 is A; The nucleotide sequence of probe 19 is sequence 19 and the nucleotide of SNP site 21 is A; The nucleotide sequence of probe 20 is sequence 20 and the nucleotide of SNP site 22 is T; The nucleotide sequence of probe 21 is sequence 21 and the nucleotide of SNP site 23 is A; The nucleotide sequence of probe 22 is sequence 22 and the nucleotide of SNP site 24 is T; The nucleotide sequence of probe 23 is sequence 23 and the nucleotide of SNP site 25 is T; The nucleotide sequence of probe 24 is sequence 24 and the nucleotide of SNP site 26 is T; The nucleotide sequence of probe 25 is sequence 25, the nucleotide of SNP site 27 is A, and the nucleotide of SNP site 28 is T. The nucleotide sequence of probe 26 is sequence 26 and the nucleotide of SNP site 29 is T; The nucleotide sequence of probe 27 is sequence 27 and the nucleotide of SNP site 30 is T; The nucleotide sequence of probe 28 is sequence 28 and the nucleotide of SNP site 31 is T.

10. The use of the specific probe of claim 8 or 9 in any of the following: (1) Prepare products for identification or auxiliary identification of Brucella bovis; (2) Prepare products for screening or breeding bovine Brucella.

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