Methods and compositions related to cooperative primers and reverse transcription
US-2024182992-A1 · Jun 6, 2024 · US
Primer group for simultaneous detection of 15 porcine pathogens through high-throughput targeted amplicon sequencing and use thereof
US12227811B2 · US · B2
| Field | Value |
|---|---|
| Publication number | US-12227811-B2 |
| Application number | US-202318509322-A |
| Country | US |
| Kind code | B2 |
| Filing date | Nov 15, 2023 |
| Priority date | Nov 18, 2022 |
| Publication date | Feb 18, 2025 |
| Grant date | Feb 18, 2025 |
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Abstract
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A primer group for simultaneous detection of 15 porcine pathogens through high-throughput targeted amplicon sequencing and use thereof are provided. The primer group for simultaneous detection of 15 porcine pathogens through high-throughput targeted amplicon sequencing includes: primers having nucleotide sequences set forth in SEQ ID NO: 1 to SEQ ID NO: 302. The nucleotide sequences of pathogenic target genes are set forth in SEQ ID NO: 303 to SEQ ID NO: 344. The primer group of the present invention is used for amplicon sequencing detection analysis method for 42 target genes of 15 porcine respiratory pathogens, has the technical advantages of rapidness, high efficiency, high targeting, strong specificity, and the like, and is suitable for rapid and accurate simultaneous diagnosis of multiple pathogens of diseased pigs in pig farms.
First claim
Opening claim text (preview).
What is claimed is: 1. A primer group for a simultaneous detection of 15 porcine pathogens through high-throughput targeted amplicon sequencing, wherein the primer group is configured to amplify pathogenic target genes, the pathogenic target genes are as follows: Pathogen name Target gene Sequence No. African swine fever ASFV-p72 SEQ ID NO: 303 virus ASFV-p54 SEQ ID NO: 304 ASFV-EP402R SEQ ID NO: 305 ASFV-MGF505-1R SEQ ID NO: 306 ASFV-MGF505-2R SEQ ID NO: 307 ASFV-MGF360-13L SEQ ID NO: 308 ASFV-MGF360-14L SEQ ID NO: 309 ASFV-K205R SEQ ID NO: 310 Porcine reproductive PRRSV-NA-orf6/M SEQ ID NO: 311 and respiratory PRRSV-NA-orf7/N SEQ ID NO: 312 syndrome virus PRRSV-EU-orf6/M SEQ ID NO: 313 PRRSV-EU-orf7/N SEQ ID NO: 314 Pseudorabies virus PRV-gE SEQ ID NO: 315 PRV-gB SEQ ID NO: 316 Porcine circovirus PCV2-cap SEQ ID NO: 317 PCV2-rep SEQ ID NO: 318 PCV3-cap SEQ ID NO: 319 PCV3-rep SEQ ID NO: 320 Classical swine fever CSFV-5′UTR SEQ ID NO: 321 virus CSFV-E2 SEQ ID NO: 322 Foot and mouth FMDV-3D-1C2B(polyprotein) SEQ ID NO: 323 diseases virus FMDV-5′UTR SEQ ID NO: 324 Swine influenza A swIAV-M SEQ ID NO: 325 virus swIAV-H1 SEQ ID NO: 326 swIAV-H3 SEQ ID NO: 327 Haemophilus parasuis HPS-vtaA SEQ ID NO: 328 HPS-infB SEQ ID NO: 329 Pasteurella multocida Pm-plpE SEQ ID NO: 330 Pm-kmt1 SEQ ID NO: 331 Actinobacillus APP-omlA SEQ ID NO: 332 pleuropneumoniae APP-apxIVA SEQ ID NO: 333 Streptococcus suis S.suis -gdh SEQ ID NO: 334 S.suis -recN SEQ ID NO: 335 S.suis -gapdh SEQ ID NO: 336 Eperythrozoon suis E.suis -ppa SEQ ID NO: 337 E.suis -g1 SEQ ID NO: 338 Toxoplasma gondii T.gondii -B1 SEQ ID NO: 339 T.gondii -RE SEQ ID NO: 340 Mycoplasma hyorhinis Mhr-p37 SEQ ID NO: 341 Mhr-16SrRNA SEQ ID NO: 342 Mycoplasma Mhp-p36 SEQ ID NO: 343 hyopneumoniae Mhp-p102 SEQ ID NO: 344; nucleotide sequences of the pathogenic target genes are set forth in SEQ ID NO: 303 to SEQ ID NO: 344; the primer group comprises a first primer group and a second primer group, wherein the first primer group comprises all odd numbered sequence identifier numbers within SEQ ID NO: 1 to SEQ ID NO: 302, an adapter sequence set forth in SEQ ID NO: 345 is added at a 5′-end of each of forward primers, the second primer group comprises all even numbered sequence identifier numbers within SEQ ID NO: 1 to SEQ ID NO: 302, and an adapter sequence set forth in SEQ ID NO: 346 is added at a 5′-end of each of reverse primers. 2. A multiplex PCR reagent for a simultaneous detection of 15 porcine pathogens through high-throughput targeted amplicon sequencing, comprising a first PCR reagent and a second PCR reagent, wherein the first PCR reagent comprises the first primer group of the primer group according to claim 1 , and the second PCR reagent comprises the second primer group of the primer group according to claim 1 , wherein each primer in the first primer group has a concentration of 10 nmol/μL in the first PCR reagent, and each primer in the second primer group has a con
Assignees
Inventors
Classifications
Primer sets for multiplex assays · CPC title
for detection or identification of organisms · CPC title
involving nucleic acid arrays, e.g. sequencing by hybridisation · CPC title
Nucleic acid amplification reactions · CPC title
for bacteria · CPC title
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- What does patent US12227811B2 cover?
- A primer group for simultaneous detection of 15 porcine pathogens through high-throughput targeted amplicon sequencing and use thereof are provided. The primer group for simultaneous detection of 15 porcine pathogens through high-throughput targeted amplicon sequencing includes: primers having nucleotide sequences set forth in SEQ ID NO: 1 to SEQ ID NO: 302. The nucleotide sequences of pathogen…
- Who is the assignee on this patent?
- Univ China Agricultural
- What technology area does this patent fall under?
- Primary CPC classification C12Q1/701. Mapped technology areas include Chemistry & Metallurgy.
- When was this patent published?
- Publication date Tue Feb 18 2025 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
- What related patents are in patentsdb?
- We list 8 related publications on this page (citations in our corpus or others sharing the same primary CPC).