An Updated TaqMan Real-time PCR for Canine and Feline Parvoviruses

Overview

Journal J Virol Methods

Specialty Microbiology

Date 2013 May 18

PMID 23680092

Citations 19

Authors

Andre Felipe Streck

Dana Ruster

Uwe Truyen

Timo Homeier

Affiliations

Soon will be listed here.

Abstract

Canine parvovirus type 2 (CPV-2) emerged in late 1970s from the feline panleukopenia virus (FPLV) and developed, since then, into novel genetic and antigenic variants (CPV-2a, -2b and -2c). Canine and feline parvoviruses cause an acute enteric disease in their hosts, with high level of viral shedding. In this study, a quantitative TaqMan PCR for detection and quantitation of canine and feline parvoviruses in serum and fecal samples was developed. The primers were designed based upon the entire GenBank content for CPV and FPLV. A standard curve was generated, and validation tests were performed using 10-fold serial dilutions of CPV-2 virus in CPV/FPLV-negative feces and CPV/FPLV-negative serum samples. As a result, the 100% detection limit of the PCR was 18 copies of the viral genome per μl of serum and fecal sample. All canine parvovirus types as well as FPLV were detected. In conclusion, the real-time PCR represents an upgraded and useful tool to identify and quantify canine and feline parvoviruses in different sample matrices.

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References

Parrish C, OConnell P, Evermann J, Carmichael L . Natural variation of canine parvovirus. Science. 1985; 230(4729):1046-8. DOI: 10.1126/science.4059921. View

Buonavoglia C, Martella V, Pratelli A, Tempesta M, Cavalli A, Buonavoglia D . Evidence for evolution of canine parvovirus type 2 in Italy. J Gen Virol. 2001; 82(Pt 12):3021-3025. DOI: 10.1099/0022-1317-82-12-3021. View

Kibbe W . OligoCalc: an online oligonucleotide properties calculator. Nucleic Acids Res. 2007; 35(Web Server issue):W43-6. PMC: 1933198. DOI: 10.1093/nar/gkm234. View

Parrish C, Have P, Foreyt W, Evermann J, Senda M, Carmichael L . The global spread and replacement of canine parvovirus strains. J Gen Virol. 1988; 69 ( Pt 5):1111-6. DOI: 10.1099/0022-1317-69-5-1111. View

Carmichael L, Joubert J, Pollock R . Hemagglutination by canine parvovirus: serologic studies and diagnostic applications. Am J Vet Res. 1980; 41(5):784-91. View

Truyen U . Evolution of canine parvovirus--a need for new vaccines?. Vet Microbiol. 2006; 117(1):9-13. DOI: 10.1016/j.vetmic.2006.04.003. View

Tamura K, Dudley J, Nei M, Kumar S . MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol. 2007; 24(8):1596-9. DOI: 10.1093/molbev/msm092. View

Shackelton L, Parrish C, Truyen U, Holmes E . High rate of viral evolution associated with the emergence of carnivore parvovirus. Proc Natl Acad Sci U S A. 2005; 102(2):379-84. PMC: 544290. DOI: 10.1073/pnas.0406765102. View

Whelan J, Russell N, Whelan M . A method for the absolute quantification of cDNA using real-time PCR. J Immunol Methods. 2003; 278(1-2):261-9. DOI: 10.1016/s0022-1759(03)00223-0. View

10.

Hirasawa T, Yono K, Mikazuki K . Detection and genomic analysis of canine parvovirus by the polymerase chain reaction. Zentralbl Veterinarmed B. 1996; 43(9):545-54. DOI: 10.1111/j.1439-0450.1996.tb00352.x. View

11.

Decaro N, Elia G, Martella V, Desario C, Campolo M, Di Trani L . A real-time PCR assay for rapid detection and quantitation of canine parvovirus type 2 in the feces of dogs. Vet Microbiol. 2004; 105(1):19-28. DOI: 10.1016/j.vetmic.2004.09.018. View

12.

Strassheim M, GRUENBERG A, Veijalainen P, Sgro J, Parrish C . Two dominant neutralizing antigenic determinants of canine parvovirus are found on the threefold spike of the virus capsid. Virology. 1994; 198(1):175-84. DOI: 10.1006/viro.1994.1020. View

13.

Kumar M, Nandi S . Development of a SYBR Green based real-time PCR assay for detection and quantitation of canine parvovirus in faecal samples. J Virol Methods. 2010; 169(1):198-201. PMC: 7119530. DOI: 10.1016/j.jviromet.2010.06.007. View

14.

Schunck B, KRAFT W, Truyen U . A simple touch-down polymerase chain reaction for the detection of canine parvovirus and feline panleukopenia virus in feces. J Virol Methods. 1995; 55(3):427-33. DOI: 10.1016/0166-0934(95)00069-3. View

15.

Uwatoko K, Sunairi M, Nakajima M, Yamaura K . Rapid method utilizing the polymerase chain reaction for detection of canine parvovirus in feces of diarrheic dogs. Vet Microbiol. 1995; 43(4):315-23. DOI: 10.1016/0378-1135(94)00102-3. View

16.

Mackay I, Arden K, Nitsche A . Real-time PCR in virology. Nucleic Acids Res. 2002; 30(6):1292-305. PMC: 101343. DOI: 10.1093/nar/30.6.1292. View

17.

Mochizuki M, San Gabriel M, Nakatani H, Yoshida M, Harasawa R . Comparison of polymerase chain reaction with virus isolation and haemagglutination assays for the detection of canine parvoviruses in faecal specimens. Res Vet Sci. 1993; 55(1):60-3. DOI: 10.1016/0034-5288(93)90035-e. View