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2007 | 51 | 1 |

Tytuł artykułu

Comparison of standard polymerase chain reaction [PCR] with real-time PCR for meq gene of Marek's disease virus

Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
The aim of this study was to compare a standard polymerase chain reaction (PCR) with real-time PCR for the detection of meq gene for the Marek's disease virus. Forty-one samples were examined in the experiment. In the standard PCR, the product characteristic for field strains (1 062 bp) was obtained in 19 samples (positive samples), in 22 samples no bands on the gel were observed (negative samples). The real-time PCR detected the viral DNA in 36 samples, so in additionally 13 more samples comparing to the standard reaction. The standard PCR revealed the viral DNA in samples at a total concentration of 10-100 ng/µL. The real-time PCR detected the viral DNA in samples at a total DNA concentration of 0.01-100 ng/µL. The compatibility of reactions was 68.2%. The sensitivity of the standard PCR with reference to the real-time PCR was found to be 86%, and the specificity of both methods 100%. Compared with conventional PCR, real-time PCR is definitely more sensitive, reproducible, quick, and has a wide dynamic range. Additionally, the application of a closed system highly reduces a risk of cross-over contamination.

Wydawca

-

Rocznik

Tom

51

Numer

1

Opis fizyczny

p.3-7,fig.,ref.

Twórcy

autor
  • National Veterinary Research Institute, 24-100 Pulawy, Poland
autor
autor
autor

Bibliografia

  • 1. Abdul-Careem M.F., Hunter B.D., Nagy E., Read L.R., Sanel B., Spencer I.L., Sharif S.: Development of a real-time PCR assay using SYBR green chemistry for monitoring Marek's disease virus genome load in feather tips. J Virol Methods 2006, 133, 34-40.
  • 2. Baigent S.J., Petherbridge L., Howes K., Smith L., Currie R., Nair V.: Absolute quantitation of Marek's disease virus genome copy number in chicken feather and lymphocyte samples using real-time PCR. J Virol Methods 2005, 123, 53-64.
  • 3. Baigent S.J., Smith L.P., Nair K., Currie R.J.: Vaccinal control of Marek's disease: current challenges, and future strategies to maximize protection. Vet Immunol Pathol 2006,112, 1-2.
  • 4. Bumstead N., Sillibourne J., Rennie M., Ross N., Davison F.: Quantification of Marek's disease virus in chicken lymphocytes using the polymerase chain reaction with fluorescence detection. J Virol Methods 1997, 65, 75-81.
  • 5. Burgess S.C., Davison T.F.: A quantitative duplex PCR technique for measuring amounts of cell-associated Marek's disease virus: differences in two populations of lymphoma cells. J Virol Methods 2002, 82, 27-37.
  • 6. Dagher H., Donninger H., Hutchinson P., Ghildyal R., Bardin P.: Rhinovirus detection: comparison of real-time and conventional PCR. J Virol Methods 2004, 117, 113- 121.
  • 7. Davidson I., Smith E. J., Perl S., Malkinson M.: Application of the polymerase chain reaction for the diagnosis of natural infection of chicken and turkeys with avian oncogenic viruses. Israel J Vet Med 1995, 50, 97- 103.
  • 8. Edwards K., Logan J., Saunders N.: Real-time PCR: an essential guide. Food Microbiol 2005, 22, 267-268.
  • 9. Endoh D., Ikegawa S., Kon Y., Hayashi M., Sato F.: Expression of the endogenous Marek's disease virus ICP4 homolog (MDV ICP4) gene is enhanced in latently infected cells by transient transfection with the recombinant MDV ICP4 gene. Jpn J Vet Res 1995, 43, 109-124.
  • 10. Hennig H., Osterrieder N., Steinhardt M., Teichert H., Kirchner H., Wandinger K.: Detection of Marek's disease virus DNA in chicken but not in human plasma. J Clin Microbiol 2003, 41, 2428- 2432.
  • 11. Islam A., Harrison B., Cheetham B.F., Mahony T.J., Young P.L., Walkden-Brown S.W.: Differential amplification and quantitation of Marek's disease viruses using real-time polymerase chain reaction. J Virol Methods 2004, 119, 103-113.
  • 12. Jang H. K., Ono M., Kato Y., Tohya Y., Niikura M., Mikami T.: Identification of a potential Marek's disease virus serotype 2 glycoprotein D gene with homology to herpes simplex virus glycoprotein D. Arch Virol 1996, 141, 2207-2216.
  • 13. Kaiser P., Underwood G., Davison F.: Differential cytokine responses following Marek's disease virus infection of chickens differing in resistance to Marek's disease. J Virol 2003, 77, 762-768.
  • 14. Kozdruń W., Samorek-Salamonowicz S., Czekaj H.: Attempt to apply the polymerase chain reaction for the diagnosis of Marek's disease. Bull Vet Inst Puławy 2001, 45, 5-10.
  • 15. Levy A.M., Burgess S.C., Davidson I., Underwood G., Leitner G., Heller E.D.: Interferon-containing supernatants increase Marek's disease herpesvirus genomes and gene transcription levels, but not virion replication in vitro. Viral Immunol 2003, 16, 501-509.
  • 16. Mullis K.B., Faloona F.: Specific synthesis of DNA in vitro via a polymerase-catalysed chain reaction. Methods Enzymol 1987, 155, 335-350.
  • 17. Morsczeck C., Langendörfer D., Schierholz J.M.: A quantitative real-time PCR assay for the detection of tetR of Tn 10 in Escherichia coli using SYBR Green and the Opticon. J Biochem Biophys Methods 2004, 59, 217- 227.
  • 18. Niesters H.G.: Quantitation of viral load using real-time amplification techniques. Methods 2001, 25, 419-429.
  • 19. Pevenstein S.R., Williams R.K., McChesney D., Mont E.K., Smialek J.E., Straus S.E.: Quantitation of latent varicella-zoster virus and herpes simplex virus genomes in human trigeminal ganglia. J Virol 1999, 73, 10514 - 10518.
  • 20. Pillay A., Radebe F., Fehler G., Htun Y., Ballard R.: Comparison of a TaqMan - based real-time PCR with conventional tests for the detection of Trichomonas vaginalis. Sex Transm Infect 2006, 7, 68-72.
  • 21. Reddy S.M., Witter R.L., Gimeno I.: Development of a quantitative-competitive polymerase chain reaction assay for serotype 1 of Marek's disease virus. Avian Dis 2000, 44, 770-775.
  • 22. Scott G. H., Franklin R.: Real-time PCR: overview and applications. Surgery 2006, 139, 1-5.

Typ dokumentu

Bibliografia

Identyfikatory

Identyfikator YADDA

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