PORÓWNANIE METOD ILOŚCIOWEGO OZNACZANIA DNA CMV W PRÓBKACH MATERIAŁU KLINICZNEGO POBRANEGO OD PACJENTÓW PO PRZESZCZEPIENIU NERKI Z OBJAWOWYM I BEZOBJAWOWYM ZAKAŻENIEM CMV
[COMPARISON OF QUANTITATIVE METHODS OF DNA CMV INVESTIGATION IN CLINICAL SAMPLES OBTAINED FROM SYMPTOMATIC AND ASYMPTOMATIC PATIENTS UNDERGO RENAL TRANSPLANTATION]

Streszczenie.

Przedstawiono wyniki oznaczeń przeprowadzonych trzema metodami pozwalającymi na ilościowe określenie poziomu DNA wirusa cytomegalii (hybrydyzacja, PCR-Amplicor, PCR w czasie rzeczywistym). Na podstawie wyników badania próbek pochodzących od pacjentów po przeszczepieniu nerki z objawowym i bezobjawowym zakażeniem CMV określono poziomy odcięcia związane z wysokim prawdopodobieństwem wystąpienia objawowej formy zakażenia.

Abstract.

Cytomegalovirus infection is one of the main problem in immunocompromised patiens. Quantitative assessment of CMV load (viral load), rate of increase of load and determination of DNA level above which the likelihood of disease is high (viral load threshold for disease) have significant prognostic and therapeutic importance at transplant recipients. The aim of this work was the comparison of 3 quantitative molecular techniques and assessment the threshold for disease for each of them. The study was undertaken with 37 samples of serum and the whole from 17 renal transplant recipients. Part
of samples (n=16) comes from symptomatic patients, and were taken in period of clinical symptoms demonstration. The samples ware investigated by hybridization method (HC) performed accordingly to Hybrid the Capture procedure, PCR Amplicor test (COBAS AMPLICOR CMV the Monitor test) and real time PCR (r-t PCR). In 21 out of 37 samples DNA CMV was detected by all 3 methods, 2 samples gave concordant negative results. The CMV DNA level measured by all 3 methods was significantly higher (p<0,05; t-Student test) in samples from symptomatic patients than from asymptomatic: 4,79 versus 3,58 for HC; 3,06 versus 1,36 for PCR-Amplicor and 4,23 versus 2,88 log DNA copies/ml for r-t PCR. The threshold for disease connected with high likelihood of disease (p<0,05; Fisher test) was established at 4 log for r-t PCR method, 4,61 for hybridization and 3 log DNA CMV copies/ml for PCR Amplicor.