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2015 | 71 | 09 |

Tytuł artykułu

Możliwości rozpoznawania zakażeń Leptospira spp. u świń z użyciem reakcji łańcuchowej polimerazy (PCR)

Autorzy

Warianty tytułu

EN
Usefulness of polymerase chain reaction (PCR) in diagnosis of infections caused by Leptospira spp. in swine)

Języki publikacji

PL

Abstrakty

EN
Leptospirosis can be an important problem and a cause of significant economic losses in swine herds. Because of a high susceptibility of leptospires to many factors, laboratory diagnosis of the disease (especially pathogen isolation and identification) is difficult and often based on serological methods. The aim of this study was to show the possible use of PCR for detection and partial identification of Leptospira spp. in clinical samples from swine. Four aborted fetuses and 2 serum samples from sows reared on a farm infected by the serovar Pomona and 4 fetuses from a farm infected by leptospires from the serogroup Sejroe were submitted for examination. Additionally, urine and serum samples from 8 aborting sows reared on 2 farms infected by the serogroup Sejroe were investigated. Serum samples were examined by the microagglutination test (MAT). Samples of urine and tissue samples from fetuses were examined by PCR with pairs of primers detecting DNA sequences specific to a) genus Leptospira, b) species L. borgpetersenii, c) two selected groups of serovars of the species L. interrogans, d) serogroup Sejroe. Serological findings showed in all examined sows the presence of titers to the serogroup Pomona or Sejroe, depending on the serogroup causing infection on a given farm. DNA of the genus Leptospira was detected in tissues of all fetuses from sows infected by the serovar Pomona, in 3 fetuses from the farm infected by the Sejroe serogroup, and in all urine samples. The presence of the DNA sequence specific for the group of L. interrogans serovars including the serovar Pomona was found in tissues of all 4 fetuses from dams presenting titers to the serogroup Pomona. DNA of the serogroup Sejroe was detected in 6 out of 8 urine samples examined, and DNA of the species L. borgpetersenii (including the serovar Sejroe) was found in 5 urine samples. No DNA of the species L. borgpetersenii or of the serogroup Sejroe was found in tissues of fetuses from the farm infected by the serogroup Sejroe. This study demonstrated the usefulness of PCR in confirming the presence of Leptospira spp. in clinical samples from swine. Furthermore, PCR confirmed the presence of the serovar Pomona in tissues of aborted fetuses and the presence of L. borgpetersenii and/or the Sejroe serogroup in samples of urine. A conclusive evaluation of the usefulness of PCR in identifying DNA of L. borgpetersenii and the serogroup Sejroe in tissue samples requires further investigations.

Wydawca

-

Rocznik

Tom

71

Numer

09

Opis fizyczny

s.568-573,fot.,tab.,bibliogr.

Twórcy

autor
  • Zakład Higieny Żywności Pochodzenia Zwierzęcego, Państwowy Instytut Weterynaryjny - Państwowy Instytut Badawczy w Puławach, Al.Partyzantów 57, 24-100 Puławy

Bibliografia

  • 1. Adler B., Lo M., Seemann T., Murray G.: Pathogenesis of leptospirosis: The influence of genomics. Vet. Microbiol. 2011, 153, 73-81.
  • 2. Bolin C.: Leptospirosis, [w:] Manual of Diagnostic Tests and Vaccines for Terrestrial Animals (mammals, birds and bees). OIE World Organisation for Animal Health, Paris, France 2012, 209-221.
  • 3. Bofim M., Barbosa-Stancioli E., Koury M.: Detection of pathogenic inn urine from naturally infected cattle by nested PCR. Vet. J. 2008, 178, 251-256.
  • 4. Branger C., Blanchard B., Fillonneau C., Suard I., Aviat F., Chevallier B., Andre-Fontaine G.: Polymerase chain reaction assay specific for pathogenic Leptospira based on the gene hap1 encoding the haemolysis-associated protein-1. FEMS Microbiol. Lett. 2005, 243, 435-437.
  • 5. CaiC., Zhu Y., Zhong Y., Xin X., Jiang X., Lou X., He P., QuinJ., Zhao G., Wang S., Guo X.: Development of O-antigen gene cluster-specific PCRs for rapid typing six epidemic serogroups of Leptospira in China. BMC Microbiology 2010, 10, 67.
  • 6. Çetinkaya B., Ertaş B., Öngör H., Muz A.: Detection of Leptospira species by polymerase chain reaction (PCR) in urine of cattle. Turk. J. Vet. Anim. Sci. 2000, 24, 123-130.
  • 7. Ellis W.: Leptospirosis, [w:] Zimmerman J., Karriker L., Ramirez A., Schwartz K., Stevenson G. (wyd.): Diseases of Swine. Wiley-Blackwell 2012, 770-778.
  • 8. Ellis W., McParland, Bryson D., Cassels J.: Boars as carriers of leptospires of the Australis serogroup on farms with abortion problem. Vet. Rec. 1986, 118, 563.
  • 9. Ellis W., McParland., Bryson D., Thiermann A., Montgomery J.: Isolation of leptospires from the genital tract and kidneys of aborted sows. Vet. Rec. 1986, 118, 294-295.
  • 10. Faine S.: Leptospira and leptospirosis. CRC Press, Boca Raton 1994.
  • 11. Gerritsen M., Olyhoek T., Smits M., Bokhout B.: Sample preparation method for polymerase chain reaction-based semiquantitive detection of Leptospira borgpetersenii serovar hardjo subtype hardjobovis in bovine urine. J. Clin. Microbiol. 1991, 29, 2805-2808.
  • 12. Gravekamp C., Van de Kemp H., Franzen M., Carrington D., Schoone G., Van Eys G., Everard C., Hartskeerl R., Terpstra W.: Detection of seven species of pathogenic leptospires by PCR using two sets of primers. J. Gen. Microbiol. 1993, 139, 1691-1700.
  • 13. Heinemann M., Garcia J., Nunes C., Gregori F., Higa Z., Vasconcellos S., Richtzenhain L.: Detection and differentiation of Leptospirs spp. in bovine semen by polymerase chain reaction and restriction fragment length polymorphism. Vet. Microbiol. 2000, 73, 261-267.
  • 14. Hookey J.: Detection of Leptospiraceae by amplification of 16S ribosomal DNA. FEMS Microbiol. Lett. 1992, 90, 267-274.
  • 15. Merien F., Amouriaux P., Perolat P., Baranton G., Saint Girons I.: Polymerase chain reaction for detection of Leptospira spp. in clinical samples. J. Clin. Microbiol. 1992, 30, 2219-2224.
  • 16. Oliveira S., de Bortolanza F., Passos D., Simões Pires Neto J., Fallavena L., Weimer T.: Molecular diagnosis of Leptospira spp. in culled sows. Braz. J. Vet. Res. 2007, 44, 18-23.
  • 17. Paiva-Cardoso M., Arent Z., Gilmore C., Hartskeerl R., Ellis W.: Altodouro, a new Leptospira serovar of Pomona serogroup isolated from rodents in northern Portugal. Infect. Genet. Evol. 2013, 13, 211-217.
  • 18. Reitstetter R.: Development of species-specific PCR primer sets for the detection of Leptospira. FEMS Microbiol. Lett. 2006, 264, 31-39.
  • 19. Savio M., Rossi C., Fusi P., Tagliabue S., Pacciarini M.: Detection and identification of Leptospira interrogans serovarsby PCR coupled with restriction endonuclease analysis of amplified DNA. J. Clin. Microbiol. 1994, 32, 935-941.
  • 20. Tramuta C., Lacerenza D., Zoppi S., Goria M., Dondo A., Ferroglio E., Nebbia P., Rosati S.: Developement of a set of multiplex standard polymerase chain reaction assays for the identification of infectious agents from aborted bovine clinical samples. J. Vet. Diagn. Invest. 2011, 23, 657-664.
  • 21. Vitale M., Vitale F., Di Marco V., Curró V., Vesco G., Caracappa S.: Polymerase chain reaction method for lrpospirosis, analysis on samples from an autochthon swine population in Sicily, Italy. Rev. Cubana Med. Trop. 2005, 57, 25-27.
  • 22. Wasiński B.: Occurrence of Leptospira sp. antibodies in swine in Poland. Bull. Vet. Inst. Pulawy 2007, 51, 225-228.
  • 23. Wasiński B.: Występowanie zakażeń bakteriami z rodzaju Leptospira u świń w latach 2002-2003. Med. Weter. 2005, 61, 45-49.
  • 24. Wasiński B., Pejsak Z.: Occurrence of leptospiral infections in swine population in Poland evaluated by ELISA and microscopic agglutination test. Pol. J. Vet. Sci. 2010, 13, 695-699.
  • 25. Woodward M., Redstone J.: Differentiation of Leptospira serovars by polymerase chain reaction and restriction fragment length polimorphism. Vet. Rec. 1993, 132, 325-326.

Typ dokumentu

Bibliografia

Identyfikatory

Identyfikator YADDA

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