US 12,227,811 B2
Primer group for simultaneous detection of 15 porcine pathogens through high-throughput targeted amplicon sequencing and use thereof
Shaolin Wang, Beijing (CN); Yang Zuo, Beijing (CN); Zhangqi Shen, Beijing (CN); Lei Zhou, Beijing (CN); Jun Han, Beijing (CN); and Jianzhong Shen, Beijing (CN)
Assigned to China Agricultural University, Beijing (CN)
Filed by China Agricultural University, Beijing (CN)
Filed on Nov. 15, 2023, as Appl. No. 18/509,322.
Claims priority of application No. 202211450766.0 (CN), filed on Nov. 18, 2022.
Prior Publication US 2024/0167106 A1, May 23, 2024
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6874 (2018.01); C12Q 1/6888 (2018.01); C12Q 1/70 (2006.01); C12Q 1/6844 (2018.01)
CPC C12Q 1/701 (2013.01) [C12Q 1/6874 (2013.01); C12Q 1/6888 (2013.01); C12Q 1/6844 (2013.01); C12Q 2600/16 (2013.01)] 5 Claims
OG exemplary drawing
 
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.