[Evaluation of the usefulness of various PCR method variations and nucleic acid hybridization for CMV infection in immunosuppressed patients].
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(0) particles of viral DNA in nested PCR to 10(2) in single PCR. The producer claimed the sensitivity of the hybridisation test to be 3 x 10(5) 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 immunossuppressed patients and in following the efficacy of antiviral treatment.
Siennicka
J
J
Zakład Wirusologii PZH w Warszawie.
Trzcińska
A
A
Litwińska
B
B
Durlik
M
M
Seferyńska
I
I
Pałynyczko
G
G
Kańtoch
M
M
D003586:Cytomegalovirus Infections / Q000175:diagnosis* / Q000276:immunology; D004279:DNA, Viral / Q000032:analysis; D006801:Humans; D016867:Immunocompromised Host / Q000276:immunology*; D007165:Immunosuppression Therapy / Q000009:adverse effects*; D016030:Kidney Transplantation / Q000276:immunology; D007938:Leukemia / Q000276:immunology; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*
D004279:DNA, Viral