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2014 | 58 | 4 |
Tytuł artykułu

Application of quantitative PCR for detection of Mycoplasma suis in blood samples

Warianty tytułu
Języki publikacji
EN
Abstrakty
EN
The aim of the study was to develop and validate a real-time PCR method, using a TaqMan probe, for quantification of Mycoplasma suis in porcine blood. No PCR signals with closely related non-haemotrophic mycoplasmas were obtained. The detection limit of PCR for plasmid combined with blood DNA was determined to be 10³/reaction (5 µL of DNA) (1.2x10⁵ target copies in 1 mL of blood). The linearity of real-time PCR (near 1) indicates its use as a quantitative method. Real-time and quantitative PCR were sensitive and specific for the detection and quantification of M. suis in the blood of animals with acute and chronic form of eperythrozoonosis. Developed quantitative PCR cannot be used to detect carrier animals with a small amount of M. suis in their blood. The validity of real-time PCR used in the studies was confirmed by the low inter- and intra-assay coefficients of variation. This fact confirms the applicability of the assay in other laboratories.
Słowa kluczowe
Wydawca
-
Rocznik
Tom
58
Numer
4
Opis fizyczny
p.533-539,fig.,ref.
Twórcy
autor
  • Department of Swine Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland
autor
  • Department of Swine Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland
autor
  • Department of Swine Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland
autor
  • Department of Swine Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland
autor
  • Department of Swine Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland
autor
  • Department of Swine Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland
autor
  • Department of Swine Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland
autor
  • Department of Swine Diseases, National Veterinary Research Institute, 24-100 Pulawy, Poland
Bibliografia
  • 1. Biondo A.W.: A review of the occurrence of hemoplasmas (hemotrophic mycoplasmas) in Brazil. Rev Bras Parasitol Vet 2009, 18, 1-7.
  • 2. Guimaraes A.M., Biondo A.W., Lara A.C., Messick J.B.: Exploratory study of Mycoplasma suis (Eperythrozoon suis) on four commercial pig farms in southern Brazil. Vet Ree 2007, 160, 50-53.
  • 3. Guimaraes A.M., Santos A.P., SanMiguel P., Walter T., Timenetsky J., Messick J.B.: Complete genome sequence of Mycoplasma suis and insights into its biology and adaption to an erythrocyte niche. PLoS One 6, 19574.
  • 4. Guimaraes A.M., Vieira R.F., Poletto R., Vemulapalli R., Santos A.P., de Moraes W., Cubas Z.S., Santos L.C., Marchant- Forde J.N., Timenetsky J., Biondo A.W., Messick J.B.: A quantitative TaqMan PCR assay for the detection of Mycoplasma suis. J Appl Microbiol 2011, 111, 417-425.
  • 5. Gwaltney S.M., Hays M.P., Oberst R.D.: Detection of Eperythrozoon suis using the poly merase chain reaction. J Vet Diagn Invest 1993, 5, 40-46.
  • 6. Henderson J.P., O'Hagan J., Hawe S.M., Pratt M.C.: Anaemia and low viability in piglets infected with Eperythrozoon suis. Vet Ree 1997, 140, 144-146.
  • 7. Hoelzle K., Grimm J., Ritzmann M., Heinritzi K., Torgerson P., Hamburger A., Wittenbrink M.M., Hoelzle L.E.: Use of recombinant antigens to detect antibodies against Mycoplasma suis, with correlation of serological results to hematological findings. Clin Vaccine Immunol 2007, 14, 1616-1622.
  • 8. Hoelzle L.E.: Haemotrophic mycoplasmas: recent advances in Mycoplasma suis. Vet Microbiol 2008. 130, 215-226.
  • 9. Hoelzle L.E.: Significance of haemotrophic mycoplasmas in veterinary medicine with particular regard to the Mycoplasma suis infection in swine. Berl Munch Tierarztl Wochenschr 2007, 120, 34-41.
  • 10. Hoelzle L.E., Adelt D., Hoelzle K., Heinritzi K., Wittenbrink M.M.: Development of a diagnostic PCR assay based on novel DNA sequences for the detection of Mycoplasma suis (Eperythrozoon suis) in porcine blood. Vet Microbiol 2003, 93, 185-196.
  • 11. Hoelzle L.E., Felder K.M., Hoelzle K.: Porcine eperythrozoonosis: from Eperythrozoon suis to Mycoplasma suis. Tierarztl Prax Ausg G Grosstiere Nutztiere 2011, 39, 215-220.
  • 12. Hoelzle L.E., Helbling M.,. Hoelzle K., Ritzmann M., Heinritzi K., Wittenbrink M.M.: First lightcycler real-time PCR assay for the quantitative detection of Mycoplasma suis in clinical samples. J Microbiol Methods 2007, 70, 346-354.
  • 13. Kutyavin I.V., Afonina I.A., Mills A., Gorn V.V., Lukhtanov E.A., Belousov E.S., Singer M.J., Walburger D.K., Lokhov S.G., Gall A.A., Dempcy R., Reed M.W., Meyer R.B., Hedgpeth J.: 3'-minor groove binder-DNA probes increase sequence specificity at PCR extension temperatures. Nucleic Acids Res 2000, 28, 655-661.
  • 14. Messick J.B., Cooper S.K., Huntley M.: Development and evaluation of a polymerase chain reaction assay using the 16S rRNA gene for detection of Eperythrozoon suis infection. J Vet Diagn Invest 1999, 11, 229-236.
  • 15. Ritzmann M.: Prevalence of Mycoplasma suis in slaughter pigs, with correlation of PCR results to hematological findings. Vet Microbiol 2009, 133, 84-91.
  • 16. Watanabe Y., Fujihara M., Suzuki J., Sasaoka F., Nagai K., Harasawa R.: Prevalence of swine hemoplasmas revealed by real-time PCR using 16S rRNA gene primers. J Vet Med Sci 2012, 74, 1315-1318.
  • 17. Wu J., Yu J., Song C., Sun S., Wang Z.: Porcine eperythrozoonosis in China. Ann N Y Acad Sci 2006, 1081, 280-285.
  • 18. Yuan C.L., Liang A.B., Yao C.B., Yang Z.B., Zhu J.G., Cui L., Yu F., Zhu N.Y., Yang X.W., Hua X.G.: Prevalence of Mycoplasma suis (Eperythrozoon suis) infection in swine and swine-farm workers in Shanghai, China. Am J Vet Res 2009, 70, 890-894.
Typ dokumentu
Bibliografia
Identyfikatory
Identyfikator YADDA
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