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Differential and Real-time Detection of Two Genotypes Porcine Circovirus Type 2 ...  

Update Date: [2013-11-14]
No.:45

Reporton.
AHRI report No.45
Topic
Differential and Real-time Detection of Two Genotypes Porcine Circovirus Type 2 (PCV2) by Loop-Mediated Isothermal Amplification Assay (LAMP)
Department
1Animal Health Research Institute, Council of Agriculture, Executive Yuan
2Graduate Institute of Veterinary Medicine, National Taiwan University
Author
C Wang1.2*, CR Jeng2, TS Huang1, VF Pang2, WM Chang1, SS Lai2, SH Hsiao2
Summary
PCV2 causes on infection in pigs is one of most clinically important swine viral diseases throughout the world.  Studies indicate that loop-mediated isothermal amplification (LAMP) is a specific, efficiency, and easy method for the detection the nucleic acids of various microorganisms in field samples through the use of 2 or 3 sets of LAMP primers.  In this study, we designed 2 sets of primers for use in a real-time PCV2 LAMP system to detect PCV2 in field samples.  To determine the diagnostic specificity of the PCV2 LAMP assay, a total of 19 PCV2 isolates, 6 porcine circovirus type 1 (PCV1) isolates, 3 pseudorabies virus (PRV) isolates, 7 porcine reproductive and respiratory virus (PRRSV) isolates, and 3 classical swine fever virus (CSFV) isolates were tested.  The results showed that only the PCV2 isolates were detected by PCV2 LAMP, without cross-reaction with other tested viruses.  The sensitivity of LAMP for the PCV2 was tested to be 10 copy numbers / reaction for positive recombinant plasmid.  These results showed that the PCV2 can be detected and distinguished accurately by using PCV2 LAMP assay with real-time monitoring.  The PCV2 LAMP is a specific, sensitive, and quick diagnostic method for the detection of PCV2.
Keyword
Porcine circovirus type 2, Loop-mediated isothermal amplification, Specificity, sensitivity