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Journal of Clinical Microbiology, September 2000, p. 3474-3477, Vol. 38, No. 9
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Quantitative, Competitive PCR Analysis of Porcine Circovirus DNA in Serum from Pigs with Postweaning Multisystemic Wasting Syndrome

Qiang Liu,* Li Wang,dagger Phil Willson, and Lorne A. Babiuk

Veterinary Infectious Disease Organization, University of Saskatchewan, Saskatoon, Saskatchewan, Canada S7N 5E3

Received 24 February 2000/Returned for modification 3 May 2000/Accepted 22 June 2000

A competitive PCR (cPCR) assay was developed for monitoring porcine circovirus (PCV) DNA in serum samples from piglets. The cPCR was based on competitive coamplification of a 502- or 506-bp region of the PCV type 1 (PCV1) or PCV2 ORF2, respectively, with a known concentration of competitor DNA, which produced a 761- or 765-bp fragment, respectively. The cPCR was validated by quantification of a known amount of PCV wild-type plasmids. We also used this technique to determine PCV genome copy numbers in infected cells. Furthermore, we measured PCV DNA loads in clinical samples. More than 50% of clinically healthy piglets could harbor both types of PCV. While PCV1 was detected in only 3 of 16 pigs with postweaning multisystemic wasting syndrome (PMWS), all the sick piglets contained PCV2. A comparison of the PCV2 DNA loads of healthy and sick animals revealed a significant difference, indicating that the development of PMWS may require a certain amount of PCV2.

* Corresponding author. Mailing address: VIDO, 120 Veterinary Rd., Saskatoon, Saskatchewan, Canada S7N 5E3. Phone: 1-306-966 7483. Fax: 1-306-966 7478. E-mail: liuq{at}sask.usask.ca.

dagger Present address: American Animal Health, Inc., Grand Prairie, TX 75052.

Journal of Clinical Microbiology, September 2000, p. 3474-3477, Vol. 38, No. 9
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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