CN115772562A - Detection kit for SNP (single nucleotide polymorphism) locus of cat genetic disease - Google Patents
Detection kit for SNP (single nucleotide polymorphism) locus of cat genetic disease Download PDFInfo
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Abstract
The invention relates to a cat genetic disease SNP locus detection kit, which belongs to the technical field of biology, and develops a fluorescence kit for simultaneously detecting 10 SNP loci by combining a fluorescence ARMS-PCR method with a capillary electrophoresis detection technology, and is used for qualitatively detecting 10 SNP loci related to five highly-developed genetic diseases of cats in vitro in genomic DNA of a cat oral swab; the template has wide application range, and blood, oral swab, umbilical cord and other cat tissue samples can be detected; the system has high specificity, good stability and high accuracy; after repeated verification, the signal strength is stable; the sensitivity is high, and accurate typing can be obtained by the amount of the DNA template as low as 0.5 ng/. Mu.L.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a cat genetic disease SNP locus detection kit.
Background
The system is used for qualitatively detecting 10 SNP sites related to five highly-developed genetic diseases of cats in the genomic DNA of the cat oral swab in vitro, and assists clinical diagnosis and accurate breeding of cats: judging whether the cat has the five highly-developed genetic diseases of the cat or not, and simultaneously detecting whether the cat carries pathogenic genes of the five highly-developed genetic diseases of the cat or not, so that accurate breeding can be guided, and the existence probability of the pathogenic genes in the cat is gradually reduced;
in order to make pet cats have a more attractive appearance, many breeds of cats are the products of inbreeding, which in turn leads to a high probability of suffering from certain genetic diseases, such as polycystic kidney disease, congenital vision disorders, congenital heart disease, etc. When the breeder carries the mutant gene, the prevalence of the offspring will be greatly increased, ultimately affecting the health and longevity of the cat. In order to improve the survival rate and the life quality of cats and avoid the congenital genetic disease of offspring caused by close breeding, it is very necessary to perform gene diagnosis before the breeding of parents.
SNP refers to Single Nucleotide Polymorphisms, and refers to variation of a Single Nucleotide on a genome, including transition, transversion, deletion and insertion. Studies have shown that some SNP mutations can lead to genetic diseases in cats. There are many methods for detecting SNPs, including direct DNA sequencing, amplification mutation system (ARMS), etc., in which direct DNA sequencing is the gold standard for mutation detection, but is very limited in detection due to its long time and high cost.
Disclosure of Invention
In order to solve the detection of gene polymorphism, the invention aims to provide a kit for detecting gene mutation based on the combination of fluorescence ARMS-PCR technology and capillary electrophoresis, the kit adopts a method of combining fluorescence PCR amplification (QF-PCR) with capillary electrophoresis detection, a common primer and a fluorescence primer are arranged for each detection site in an amplification system, the primers are combined with a corresponding genomic DNA template and amplified to generate an amplification product with a specific length and a fluorescence label, the amplification product is detected by capillary electrophoresis, and the genotype of a sample is determined according to the sizes of different fluorescence label fragments and the ratio of the peak heights of the products of gene sites.
The gene mutation mainly comprises the following steps: point mutations, deletion mutations and insertion mutations. The deletion mutation in the present invention may include a mutation of deleting one or more bases, and the insertion mutation may also include a mutation of inserting one or more bases.
The invention integrates ARMS-PCR technology and capillary electrophoresis detection technology. The method is characterized in that in the same system, the wild template ARMS primer, the mutant template ARMS primer and the upstream or downstream primer corresponding to the ARMS primer are used, a reaction tube is used for detecting 10 SNP of the same sample to be detected, and the genotype of the sample is determined according to the fragment size and the product peak height ratio of the gene locus.
According to the ARMS-PCR technical principle, the mismatched bases at the 3' end region of the ARMS primer are increased, so that the wild type ARMS primer does not amplify a mutant sample or has low amplification efficiency; and the ARMS primers aiming at the mutant template do not amplify or have low amplification efficiency on the wild sample, and the specificity of the primers is enhanced, so that the result is easier to judge.
The kit provided by the invention is directed to point mutation, and comprises: the ARMS primer aiming at the wild type template, wherein the 3 'end of the primer is terminated at the position of a site to be detected and matched with the wild type sequence, and mismatched bases are added at the positions of bases 2 to 4 at the 3' end of the wild type ARMS primer;
ARMS primers for a mutant template, wherein the 3' end of the primers is terminated at a site to be detected and matched with a mutant sequence, mismatched bases are added at the positions of 2-4 bases at the 3' end of the mutant ARMS primers, and 4 bases are added at the 5' end of the mutant ARMS primers;
sharing an upstream or downstream fluorescent primer, and amplifying a sequence containing a mutation site of a gene to be detected together with a wild type ARMS primer and a mutant ARMS primer;
the kit of the invention is directed to insertion or deletion mutations, the kit comprising: designing a pair of common amplification primers, adding fluorescence modification at the 5' tail end of the upstream or downstream of the primers, detecting by capillary electrophoresis, and distinguishing the type of a sample to be detected according to the size of a fragment.
In the present invention, the kit further comprises: 2.5 × Master Mix (main components: dNTPs, polymerase, mg2+, etc.), DNA quality control 1 (wild type template), DNA quality control 2 (mutant template), nuclease-free pure water and capillary electrophoresis detection internal standard QD550.
In the kit of the present invention, the 5 'end of the fluorescent primer is fluorescently labeled, for example, the primer is labeled with FAM or HEX at the 5' end.
The invention also aims to develop a method for simultaneously detecting 10 SNP mutation sites in one tube based on the combination of a fluorescence ARMS-PCR method and a capillary electrophoresis detection technology. The method comprises the following steps: in a reaction system (including sample DNA,2.5 × Master Mix, and no nuclease pure water), the primer mixed solution is used for detecting the same sample to be detected, and 10 SNP genotypes of the sample are determined according to different sizes of fluorescence labeling fragments and the ratio of the product peak heights of gene loci.
The technical scheme of the invention comprises the following design ideas:
(1) ARMS primer design: the primers are designed by software such as Primer Premier5, NCBI Blast and the like, the Tm value of each Primer is ensured to be in the range of (60 +/-2) DEG C as much as possible when the primers are designed, the amplification efficiency is similar, and the difference of the sizes of amplification products of each pair of primers is ensured to be more than 10 bp. After the design is finished, software such as AutoDimer and the like is used for analyzing the interaction between the primer dimer and different primers, and if the interaction exists, a non-specific product or dimer can be generated and redesigned until a primer sequence meeting the requirement is obtained.
(2) DNA extracted from a cat mouth swab and a reference substance are selected as templates, the 10 pairs of primers are respectively used for carrying out single amplification, and a PCR system and amplification conditions are adjusted through a capillary electrophoresis result to obtain common amplification conditions of the 10 pairs of primers. Through multiple rounds of adjustment, the balance, efficiency and miscellaneous peak condition of the system are integrated, and the amplification effect is optimal when the system is at 59 ℃, so 59 ℃ is selected as the annealing temperature for amplification of the kit.
(3) Constructing a composite system: putting 10 pairs of primers into the same tube according to the same proportion for amplification, adjusting respective concentration according to the capillary electrophoresis result of the composite amplification, ensuring that the amplification efficiency (reaction is on the peak height of the electrophoresis result) of each primer pair is basically consistent and the detection result of the same SNP locus has no interference of a foreign peak, and redesigning the primers if the primers can not meet the conditions. Finally, 10 pairs of ARMS primer sequences are determined.
(4) Performance study: and selecting a mutant sample screened by the kit amplification detection, performing amplification sequencing on the sample by using a gold standard sanger sequencing method, comparing the sample with a detection result of the kit, verifying the detection result of the kit, and confirming the accuracy of the kit.
(5) Positive judgment value: selecting a part of representative typing samples, and determining the genotyping of each locus of the samples by using a 'gold standard' method (sequencing); the kit is used for detecting according to a standard experimental process, and the kit result is compared with a gold standard result; the detection result of the kit is statistically analyzed by using Excel to determine the size of the fragment and the range of the peak height, and SPSS 18.0 is used for drawing a Receiver Operating Characteristic (ROC) curve to determine the range of the ratio of the peak height and the range of the effective peak of the recombined sample, namely the result interpretation standard.
According to the ROC curve result, the cut-off value of the peak height ratio of the wild type/insertion type and the heterozygous type is determined to be 5.12, the cut-off value of the peak height ratio of the homozygous mutant type/deletion type and the heterozygous type is determined to be 0.11, and the typing threshold value parameters are determined as shown in the following table.
Typing threshold parameter
| Ratio | Sample typing |
| <0.11 | Homozygous mutant/deletion type |
| 0.11～5.12 | Heterozygote type |
| >5.12 | Wild type/insertion type |
(6) Minimum detection limit: in this study, genomic DNA extracted from cat buccal swabs was diluted with a concentration gradient, and the assay was repeated 3 times at each concentration level, with the lowest detectable concentration of 100% being the lowest detection limit. And respectively carrying out amplification tests on samples with three concentration gradients of 0.2 ng/. Mu.L, 0.5 ng/. Mu.L and 1 ng/. Mu.L, detecting an amplification product by using ABI 3500xl Dx, and groping the detection sensitivity of the cat SNP detection kit. In order to guarantee the detection rate of more than 95% of the subsequent detection rate, the minimum detection rate of the DNA sample is finally limited to 0.5 ng/mu L.
(7) And (3) repeatability verification: the kit is used for carrying out repeated amplification test on a sample with the concentration of 1.5 ng/. Mu.L, and ABI 3500xl Dx is used for detecting an amplification product, so that the repeatability of the kit is tested. Through experimental data analysis, the peak types of all SNP sites in 3 times of repetition can be clearly detected, and the typing is consistent, so that the kit can be judged to have good repeatability.
(8) The cat SNP detection kit is subjected to amplification test by using ABI9700, ABI Veriti 96 and ABI Veriti DX 96PCR amplification instruments, products are detected by using ABI 3500XL DX and ABI3730 gene detectors respectively, and the applicable model of the cat SNP detection kit is explored. Through data analysis, the amplification efficiency of ABI9700 is slightly higher than that of ABI Verti 96 and ABI Verti DX 96, and the detection efficiency of ABI 3500XL DX is slightly higher than that of ABI 3730; the system can be normally amplified under the conditions of ABI9700, ABI Verti 96 and ABI Verti DX 96, the performance is not influenced by an amplification instrument, amplification products can be clearly detected on 3500XL DX and ABI3730, and the detection results of different instruments are consistent.
(9) Stability study: and (3) storing the kit at the temperature of minus 20 +/-5 ℃ in the dark until the reagent is completely frozen, and then performing the next thawing and freezing process. The kit is repeatedly frozen and thawed, samples are detected through amplification, and the detection result condition of each sample under different times of repeated freezing and thawing of the kit is analyzed, so that the limitation of the times of repeated freezing and thawing of the kit is evaluated. After the kit is repeatedly frozen and thawed for 0, 2, 4, 5 and 6 times, the amplification detection is respectively repeated for 3 times on 14 samples by using the freezing and thawing kit each time, the repeated detection results are consistent, the freezing and thawing results for 5 times are consistent, the detection results are not abnormal after 6 times of freezing and thawing, and the samples can be normally typed, so that the kit can still effectively amplify the detection samples after 6 times of freezing and thawing.
(III) advantageous effects
The invention provides a kit for detecting SNP loci of cat genetic diseases. The method has the following beneficial effects:
the kit adopts a method of combining Fluorescent PCR amplification (QF-PCR) with capillary electrophoresis detection, a common primer and a Fluorescent primer are arranged for each detection site in an amplification system, and the primers are combined with corresponding genome DNA templates for amplification to generate an amplification product with a specific length and a Fluorescent label. Detecting the amplified product by capillary electrophoresis, determining the genotype of the sample by the sizes of different fluorescence labeling fragments and the ratio of the product peak heights of the gene sites, wherein the genotype contains 5 common genetic disease sites of cats, and basically covers mainstream detection sites in the market; the template has wide application range, and blood, oral swab, umbilical cord and other cat tissue samples can be detected; the system has good specificity, stability and accuracy, and the signal intensity is stable after repeated verification; the sensitivity is high, and the accurate typing can be obtained by the DNA template amount as low as 0.5 ng/. Mu.L.
Drawings
FIG. 1 kit quality control 1 amplification results
FIG. 2 kit quality control 2 amplification results
FIG. 3 kit lowest detection limit FAM channel amplification results
FIG. 4 lowest detection limit HEX channel amplification results for the kit
FIG. 5 reproducibility of FAM channel amplification results
FIG. 6 reproducibility validation HEX channel amplification results
FIG. 7ABI 9700 amplification plasmid DNA HET detection results
FIG. 8ABI Veriti 96 amplification plasmid DNA HET detection results
FIG. 9ABI Veriti DX 96 amplified plasmid DNA HET test results.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1 amplification of genomic DNA quality control product 1 extracted from oral swab of cat by Using a Cat genetic disease SNP site detection kit
(1) Collect cat oral cavity swab
(2) DNA extraction: DNA was prepared using a Tiangen "high efficiency buccal swab genomic DNA extraction kit".
(3) Reaction system: shaking and mixing each reaction reagent (2.5 times Master Mix, primer mixed solution and nuclease-free pure water), preparing PCR reaction mixed solution according to the volume ratio (except for template), subpackaging 9 microlitres in PCR reaction tubes, finally adding 1 microlitres of template into each reaction tube, centrifuging and carrying out the next step. The reaction system composition is shown in the following table.
Standard amplification reaction system
| Component name | Volume (mu L) |
| 2.5×Master Mix | 8 |
| Primer mixture | 2 |
| Nuclease-free pure water | 9 |
| |
1 |
| Total volume | 20μL |
(4) PCR reaction procedure: the PCR reaction tube was placed on the amplification instrument and the following program was designed and run: step 1: denaturation at 95 ℃ for 5min, step 2: denaturation at 97 ℃ for 10 seconds, step 3: annealing at 59 ℃ for 30 seconds, and step 4: extension at 72 ℃ for 30 seconds, repetition of 2 to 4 steps 30 times, and final extension at 72 ℃ for 10 minutes. After the PCR program is operated, the PCR product can be immediately subjected to subsequent detection; or storing at 2-8 ℃ and detecting within 24 hours; or storing at-20 deg.C, and detecting within three days. Repeated freeze thawing should be avoided as much as possible, or the number of freeze thawing times should not exceed 3
(5) And (3) capillary electrophoresis detection: mixing the QD550 internal standard and formamide according to the ratio of 2.5.
(6) And (3) data analysis: and importing original data, selecting Add sample to project from a File menu of a main page, finding a sample File, selecting a folder, clicking Add to list, clicking Add, displaying the sample File in a project window, and selecting analysis parameters. Analysis method, panel and size standard are defined. And selecting analysis > analysis selected sample in a main page menu, presenting a save project dialog box, saving the dialog box after naming, starting the software to process data, and displaying analysis completed in the lower left corner after analysis. The data obtained were analyzed and a map generated using GeneMapper software, see figure 1.
Example 2: a cat genetic disease SNP locus detection kit is applied to amplify mutant type plasmids, namely quality control products 2 in the kit, wherein the mutant type plasmids are a DNA sequence containing mutant loci and are synthesized by Shanghai bioengineering GmbH.
(1) PCR amplification was performed according to the amplification conditions in example 1
(2) The assay was performed on a genetic Analyzer as in example 1, and the data obtained were analyzed and mapped using GeneMapper software, see FIG. 2
While there have been shown and described what are at present considered the fundamental principles and essential features of the invention and its advantages, it will be apparent to those skilled in the art that the invention is not limited to the details of the foregoing exemplary embodiments, but is capable of other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (6)
1. A cat genetic disease SNP locus detection kit is characterized in that: the SNP locus is directed at point mutation, and the kit comprises:
setting an ARMS primer of a wild type template, wherein the 3 'end of the primer is terminated at the site to be detected and matched with a wild type sequence, and a mismatched base is set at the 2 nd to the 4 th positions of the 3' end of the primer;
setting ARMS primer of mutant template, wherein the 3' end of the primer is terminated at the position to be detected and matched with the mutant sequence, wherein 2-4 th position of the 3' end of the primer is set with a mismatched base, and 4 bases are added at the 5' end to distinguish fragment length;
and sharing an upstream fluorescent primer, and amplifying a sequence containing the mutation site of the gene to be detected by the fluorescent primer, the ARMS primer aiming at the wild type template and the ARMS primer aiming at the mutant type template to generate an amplification product with specific length and fluorescent label. And detecting the amplified product by capillary electrophoresis, and determining the genotype of the sample according to the size of the fragment and the ratio of the product peak height of the gene locus.
2. The kit for detecting the SNP locus of a feline genetic disease according to claim 1, characterized in that: the SNP site is directed to insertion or deletion mutation, and the kit comprises: designing a pair of amplification primers, and adding a fluorescent label to the forward or reverse 5' ends of the primers, wherein the primers together amplify a sequence containing the mutation site of the gene to be detected to generate an amplification product with a specific length and a fluorescent label. And detecting the amplified product by capillary electrophoresis, and determining the genotype of the sample according to the size of the fragment and the ratio of the product peak height of the gene locus.
3. The kit for detecting the SNP locus of a feline genetic disease according to claim 1, characterized in that: the primer sequences of 10 SNP sites are respectively as follows:
the wild primer PKD1-WR for amplifying polycystic kidney disease PKD1 (c.9882C > A) is shown as SEQ ID NO 1;
the mutant primer PKD1-MR for amplifying polycystic kidney disease PKD1 (c.9882C > A) is shown as SEQ ID NO 2;
a common primer PKD1-F for amplifying polycystic kidney disease PKD1 (c.9882C > A) is shown as SEQ ID NO3, and 5' FAM is modified;
the wild-type primer KIF3B-WR for amplifying the progressive retinal atrophy KIF3B (c.1000G > A) is shown as SEQ ID NO 4;
the mutant primer KIF3B-MR for amplifying the progressive retinal atrophy KIF3B (c.1000G > A) is shown as SEQ ID NO 5;
a common primer KIF3B-F for amplifying progressive retinal atrophy KIF3B (c.1000G > A) is shown as SEQ ID NO6, and 5' HEX is modified;
the wild-type primer CEP290-WF for amplifying the progressive retinal atrophy CEP290 (IVS 50+9T > G) is shown as SEQ ID NO 7;
the mutant primer CEP290-MF for amplifying the progressive retinal atrophy CEP290 (IVS 50+9T > G) is shown as SEQ ID NO 8;
the common primer CEP290-R of the amplified progressive retinal atrophy CEP290 (IVS 50+9T > G) is shown as SEQ ID NO9, and 5' FAM is modified;
the wild type primer MYBPC3-91-WF for amplifying hypertrophic cardiomyopathy MYBPC3 (c.91G > C) is shown as SEQ ID NO 10;
the mutant primer MYBPC3-91-MF for amplifying hypertrophic cardiomyopathy MYBPC3 (c.91G > C) is shown as SEQ ID NO 11;
a common primer MYBPC3-91-R for amplifying hypertrophic cardiomyopathy MYBPC3 (c.91G > C) is shown as SEQ ID NO12, and 5' FAM is modified;
the wild primer MYBPC3-2458-WF for amplifying hypertrophic cardiomyopathy MYBPC3 (c.248C > T) is shown as SEQ ID NO 13;
the mutant primer MYBPC3-2458-MF for amplifying hypertrophic cardiomyopathy MYBPC3 (c.248C > T) is shown as SEQ ID NO 14;
the common primer MYBPC3-2458-R for amplifying hypertrophic cardiomyopathy MYBPC3 (c.248C > T) is shown as SEQ ID NO15, and 5' HEX is modified;
the wild primer MYH7-WR for amplifying hypertrophic cardiomyopathy MYH7 (c.5647G > A) is shown as SEQ ID NO 16;
the mutant primer MYH7-MR for amplifying hypertrophic cardiomyopathy MYH7 (c.5647G > A) is shown as SEQ ID NO 17;
a common primer MYH7-F for amplifying hypertrophic cardiomyopathy MYH7 (c.5647G > A) is shown as SEQ ID NO18, and 5' FAM is modified;
the wild type primer TNNT2-WF of the amplified hypertrophic cardiomyopathy TNNT2 (c.95intron (-108) G > A) is shown as SEQ ID NO 19;
the mutant primer TNNT2-MF of the amplified hypertrophic cardiomyopathy TNNT2 (c.95intron (-108) G > A) is shown as SEQ ID NO 20;
a common primer TNNT2-R of the amplified hypertrophic cardiomyopathy TNNT2 (c.95intron (-108) G > A) is shown as SEQ ID NO21, and 5' HEX is modified;
the wild primer ALMS1-WR for amplifying hypertrophic cardiomyopathy ALMS1 (c.7384G > C) is shown as SEQ ID NO 22;
the mutant primer ALMS1-MR for amplifying hypertrophic cardiomyopathy ALMS1 (c.7384G > C) is shown as SEQ ID NO 23;
a common primer ALMS1-F for amplifying hypertrophic cardiomyopathy ALMS1 (c.7384G > C) is shown as SEQ ID NO24, and 5' HEX is modified;
amplification of pyruvate kinase deficiency PKLR
(c.682 _694delGTGCAGAAAATCG (old: c.693+304G >) primer PKLR-F is shown as SEQ ID NO25, 5' FAM is modified;
amplification of pyruvate kinase deficiency PKLR
The primer PKLR-R of (c.682 _694delGTGCAGAAAATCG (old: c.693+304G >) is shown as SEQ ID NO 26;
primers MAN2B1-F amplifying mannosidosis MAN2B1 (c.1749 _1752 delCCAG) are shown as SEQ ID NO27, 5' HEX is modified;
primers MAN2B1-R for amplifying mannosidosis MAN2B1 (c.1749-1752 delCCAG) are shown in SEQ ID NO 28.
4. The kit for detecting the SNP locus of a feline genetic disease according to claim 1, characterized in that: the kit also comprises 2.5 × Master Mix, and the main components are as follows: dNTPs, a DNA quality control product wild type template, a DNA quality control product mutant type template, nuclease-free pure water and an internal standard QD550 for capillary electrophoresis detection.
5. The kit for detecting the SNP locus of a feline genetic disease according to claim 1, characterized in that: the amplification system comprises: 8 mu L of 2.5 × Master Mix, 2 mu L of primer mixture, 9 mu L of nuclease-free pure water, 1 mu L of DNA template to be detected or 1 mu L of quality control substances 1 and 2; the reaction conditions of the amplification system are as follows: pre-denaturation at 95 ℃ for 5 min; denaturation at 97 ℃ for 10 seconds, annealing at 59 ℃ for 30 seconds, and extension at 72 ℃ for 30 seconds for 30 cycles; extension at 72 ℃ for 10 min.
6. The kit for detecting the SNP locus of a feline genetic disease according to claim 1, characterized in that: the capillary electrophoresis detection reaction system comprises: 8.8 μ L Hi-Di, 0.2 μ L QD550 internal standard, 1 μ L LPCR product stock solution, cover with a rubber pad, use a table or plate centrifuge, instantaneous centrifugation; reaction conditions are as follows: denaturation at 95 deg.C for 5min, immediately ice-cooling at-20 deg.C for more than 5min, detecting within 24 hr, instantly centrifuging with desk type or plate type centrifuge, performing capillary electrophoresis with gene analyzer, and selecting "Fragment" electrophoresis method.
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| Application Number | Priority Date | Filing Date | Title |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118745461A (en) * | 2024-08-05 | 2024-10-08 | 英科新创（苏州）生物科技有限公司 | Application and products of mutation sites of cat progressive retinal atrophy-related genes |
| CN118745460A (en) * | 2024-08-05 | 2024-10-08 | 英科新创（苏州）生物科技有限公司 | Application, reagents and diagnostic kits for cat PKD1 gene mutation sites |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118745461A (en) * | 2024-08-05 | 2024-10-08 | 英科新创（苏州）生物科技有限公司 | Application and products of mutation sites of cat progressive retinal atrophy-related genes |
| CN118745460A (en) * | 2024-08-05 | 2024-10-08 | 英科新创（苏州）生物科技有限公司 | Application, reagents and diagnostic kits for cat PKD1 gene mutation sites |
| CN118745460B (en) * | 2024-08-05 | 2025-07-11 | 英科新创（苏州）生物科技有限公司 | Application of cat PKD1 gene mutation site, reagent and diagnostic kit |
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