Ocena przydatnosci roznych wariantow metody PCR oraz metody hybrydyzacji kwasow nukleinowych w wykrywaniu zakazenia CMV u pacjentow z immunosupresja

• Autorzy: Siennicka J. , Trzcinska A. , Litwinska B. , Durlik M. , Seferynska I. , Palynyczko G. , Kantoch M.
• Języki publikacji: PL

Abstrakty

PL

Porównano diagnostyczną przydatność metody hybrydyzacji i różnych wariantów PCR do wykrywania DNA wirusa cytomegalii w próbkach krwi pacjentów z immunosupresją.

EN

In diagnosis of CMV infection various laboratory methods are used. The methods based on detection of viral nucleic acids have been introduced routinely in many laboratories. The aim of this study was to compare nucleic acid hybridisation method and various variants of PCR methods with respect to their ability to detect CMV DNA. The studied material comprised 60 blood samples from 19 patients including 13 renal transplant recipients and 6 with acute leukaemia. The samples were subjected to hybridisation (Murex Hybrid Capture System CMV DNA) and PCR carried out in 3 variants: with one pair of primers (single PCR), nested PCR and Digene SHARP System with detection of PCR product using a genetic probe in ELISA system. The sensitivity of the variants ranged from 10° particles of viral DNA in nested PCR to 102 in single PCR. The producer claimed the sensitivity of the hybridisation test to be 3 x 105 and it seems to be sufficient for detection of CMV infection. The obtained results show that sensitivity of hybridisation was comparable to that of single PCR and the possibility of obtaining quantitative results makes it superior, on efficacy of antiviral therapy. especially in monitoring CMV infection in immunossupressed patients and in following the efficacy of antiviral treatment.

Słowa kluczowe

PL

choroby wirusowe; wirusy; cytomegalia; lancuchowa reakcja polimerazy; hybrydyzacja; diagnostyka laboratoryjna; immunosupresja; DNA

• Wydawca: -
• Czasopismo: Medycyna Doświadczalna i Mikrobiologia
• Rocznik: 2000
• Tom: 52
• Numer: 3
• Strony: 283-293
• Opis fizyczny: s.283-293,fot.,rys.,tab.,bibliogr.

Twórcy

autor

Siennicka J.

• Panstwowy Zaklad Higieny, 00-791 Warszawa, ul.Chocimska 24

autor

Trzcinska A.

autor

Litwinska B.

autor

Durlik M.

autor

Seferynska I.

autor

Palynyczko G.

autor

Kantoch M.

Bibliografia

• 1. Arai S, Mangano M, Starr SE i inni. Optimalization of conditions for detecting human cytomegalovirus DNA with nested PCR. W: PCR: protocols for diagnosis of human and animal virus diseases. Red. Y Becker, G Darai, Springer-Verlag, Berlin, Heidelberg, 1995, 205-13
• 2. Bauer HM, Ting Y, Greer CE i inni. Genital human papillomavirus infections in female university students as determined by a PCR-based method. JAMA 1991; 265: 472-77.
• 3. Britt WJ. Infections associated with human cytomegalovirus. W: Herpesvirus infections. Red. R Glaser, JF Jones, Marcel Dekker, Inc, New York, Basek Hong Kong, 1994, 59-116.
• 4. Dal Monte P, Lazzarotto T, Ripalti A i inni. Human cytomegalovirus infection: a complex diagnostic problem in which molecular biology has induced a rapid evolution. Intervirology 1996; 39: 193-203.
• 5. Drouet E, Michelson S, Denoyel G i inni. Polymerase chain reaction detection of human cytomegalovirus in over 2000 blood specimens correlated with wirus isolation and related to urinary wirus excretion. J Virol Method 1993; 45: 259-76.
• 6. Grundy JE, Ehrnst A, Einsele H i inni. A three-center european external quality control study of PCR for detection of cytomegalovirus DNA in blood. J Clin Microbiol 1996; 34: 1166-70.
• 7. Gut W, Kańtoch M. Diagnostyczne badania wirusologiczne: aktualne możliwości i problemy. Post Nauk Med 1996; 9: 174-79.
• 8. Hage DS, Immunoassays. Anal Chem 1999; 71: 2948-3048.
• 9. Hibberd PL, Tolkof-Ruhin NE, Cosimi AB i inni. Symptomatic cytomegalovirus disease in the cytomegalovirus antibody seropositive renal transplant recipients treated with OKT3. Transplantation 1992; 53: 68-72.
• 10. Jarząbek Z, Gut W, Szczepaniak Z i inni. Ocena przydatności immunoenzymatycznych zestawów diagnostycznych i testów lateksowych do wykrywania rotawirusów. Med Dośw Mikrobiol 1990; 42: 54-60.
• 11. Miller MJ, Lazar J Moe AA i inni. Evaluation of Hybrid Capture System for the detection of CMV wiremia. Presented at the Clinical Virology Symposium and Annual Meeting Pan American Society for Clinical Virology, Alabama, April 1996.
• 12. Siennicka J, Litwińska B, Seferyńska I i inni. Diagnostyka zakażeń CMV u pacjentów z rozpoznaniem ostrej białaczki. Med Dośw Mikrobiol 1998; 50: 285-92.
• 13. Siennicka J, Durlik M, Litwińska B i inni. Identification of cytomegalovirus (CMV) infection by different laboratory methods in renal transplant recipients undergoing triple-drug immunosupressive treatment. Acta Microbiol Pol 1999; 48: 61-71.