Phages harboring specific peptides that recognize the N protein of the porcine reproductive and respiratory syndrome virus distinguish the virus from other viruses.
The aim of the current study was to develop a novel diagnostic test for detecting porcine reproductive and respiratory syndrome virus (PRRSV) using phage display technology. The N gene of PRRSV isolate HH08 was cloned following reverse transcription-PCR. Sequence comparison indicated that the N gene shared 96.4% homology to that of North American PRRSV (isolate VR2332) and 35.5% with that of European PRRSV (isolate LV), indicating that the PRRSV isolate was related to the North American PRRSV genotype. The bacterially expressed N protein was used as a target in a biopanning process using a phage display random peptide library. Seven phages expressing different peptides had a specific binding activity with the N protein. The putative binding motifs were identified by DNA sequencing. More importantly, the selected phages harboring specific peptides that recognize the N protein of PRRSV were able to efficiently distinguish PRRSV from other viruses in enzyme-linked immunosorbent assays.
College of Veterinary Medicine, Northeast Agricultural University, 59 Mucai St., Xiangfang District, 150030 Harbin, China.
Xiaofeng
X
Ren
Mingcui
M
Wang
Jiechao
J
Yin
Guangxing
G
Li
D000818:Animals; D001435:Bacteriophages / Q000235:genetics*; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D008969:Molecular Sequence Data; D019151:Peptide Library; D019318:Porcine Reproductive and Respiratory Syndrome / Q000175:diagnosis*; D019316:Porcine respiratory and reproductive syndrome virus / Q000302:isolation & purification*; D011485:Protein Binding; D017422:Sequence Analysis, DNA; D014764:Viral Proteins / Q000378:metabolism*; D014773:Virology / Q000379:methods*
D004279:DNA, Viral; D019151:Peptide Library; D014764:Viral Proteins