PCV2和PCV3双重纳米PCR方法的建立及初步应用

doi:10.16431/j.cnki.1671-7236.2017.12.007

Abstract

Abstract:

This study was aimed to establish a duplex nanoparticle-assisted PCR (nano-dPCR) method for the simultaneous detection of porcine circovirus type 2 (PCV2) and PCV3, and to apply this method in the detection of PCV2 and PCV3. Primers specific to PCV2 and PCV3 were designed by reference to the respective gene sequences in GenBank. The reaction conditions of nano-dPCR were also optimized. An evaluation of the specificity and sensitivity of the established nano-dPCR protocol proved the method to be both specific and sensitive, with the lower detection limit for the amount of nucleic acid in PCV2 and PCV3 to be 93.2 and 91.6 copies/μL, respectively. This sensitivity was 100 times higher than that of conventional PCR. The method was applied to inspect 265 clinical samples sent for testing, and the results showed that PCV2 and PCV3 infection in pig was rather common, with 16.6% positive for PCV3, 14.7% positive for PCV2, and 6.8% for mixed infection. The detection results on clinical samples supported the newly established nano-dPCR method as a rapid and sensitive differential diagnosis for the early infection of PCV2 and PCV3.

Key words: duplex nanoparticle-assisted PCR (nano-dPCR); porcine circovirus 2 (PCV2); porcine circovirus 3 (PCV3); pathogen detection

CLC Number:

S852.65

LIANG Lin, PANG Chun-hua, LUO Ya-kun, ZHOU Ling, WANG Jing, LIU Chang, LIU Qi, CUI Shang-jin. Establishment and Rudimentary Application of the nano-dPCR for Detection of PCV2 and PCV3[J]. , 2017, 44(12): 3440-3445.

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Establishment and Rudimentary Application of the nano-dPCR for Detection of PCV2 and PCV3

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