"pp65 antigenemia and real time polymerase chain reaction (PCR) based-study to determine the prevalence of human cytomegalovirus (HCMV) in kidney donors and recipients with follow-up studies."

Abstract

BackgroundThe present study was undertaken to determine the rate of occurrence of Human cytomegalovirus (HCMV) among kidney transplant recipients and donors by application of direct detection methods and to understand HCMV infection/disease development among transplanted patients as a prospective study.ResultsPeripheral blood samples collected from 76 kidney donors and 76 recipients from September 2007 to August 2009 were subjected to pp65 antigenemia and Quantitative real-time PCR (qRT-PCR) assays. Data were analyzed under Group A, B and C. Group A was further divided into sub-groups I, II, III, IV, and V for better understanding. Three, one and two donors in sub-group I, III, IV of Group A tested positive for real time PCR respectively. One recipient from group III tested positive for HCMV by qRT- PCR prior transplantation and remained positive one month post-transplantation. Three other recipients, tested negative prior to transplantation became positive a month after transplantation. Group B consisted of 18 donor-recipient pairs and one of the donor tested positive for HCMV by qRT-PCR. Eight recipients tested positive for HCMV one month after transplantation. The pp65 positivity and HCMV DNA load was high among group C recipients who mostly had symptoms of active disease. Significantly high values of pp65 antigenemia were observed among recipients of sub-group II (non-parametric chi-square test p = 0.007). Positive correlation between pp65 antigenemia and qRT-PCR value was observed. Thirty three of the recipients with disease treated with Valgancyclovir showed improved clinical outcome.ConclusionOur study showed that a significant proportion of kidney recipients develop HCMV infection following renal transplantation in spite of the absence of HCMV among donors. pp65 antigenemia assay and qRT- PCR methods can be applied to detect HCMV among kidney donors and recipients to monitor development of disease and these assays were predicative of HCMV infection among them. Clinical resistant to valganciclovir was not observed.

Clinical validation of an in-house quantitative real time PCR assay for cytomegalovirus infection using the 1st WHO International Standard in kidney transplant patients

“…Another study conducted in Brazil in a low-risk population of kidney transplant recipients found an incidence rate of 69.6% using antigenemia and qPCR 14 methodologies, yet a cohort study performed in heart transplant recipients found a rate of 93.3% incidence 15 . The incidence rates found in Brazil are similar to studies in Japan (70.8%) 16 and India (73.7%) 17 but differ from countries such as Korea, where the literature shows rates of 30-40% 18 - 21 , Finland of 27% 22 and in the USA, in a pediatric kidney transplant population, a rate of 27% was found. 23 …”

Determination of human cytomegalovirus pp65 antigenemia among renal transplant patients

“…[39–41] In our earlier report, we reported the rate of positivity of HCMV to be very high among the transplant patients. [27] Therefore a pp65 antigenemia count of 10 reported based on the Western literature[42–46] may not reflect the true situation of replication of HCMV in PBLs in Indian patients. It should also be borne in mind that pp65 antigenemia assay is subjective as it is prone to the visual errors of individuals reporting on the smears.…”

“…[27] The intra- and inter-assay reproducibility were evaluated using triplicates of plasmid dilutions (10 1 , 10 3 , and 10 5 ) corresponding to an input of 2.5 × 10 3 , 2.5 × 10 5 , and 2.5 × 10 7 copies/ml per reaction in the same and four independent runs, respectively. RT-PCR for the quantification of CMV was performed on the viral isolates that propagated on corneal fibroblast culture.…”

PP65 antigenemia in the diagnosis of cytomegalovirus infection in AIDS patients

“…We considered 3 x 10 5 leukocytes to be adequate for this study (8,17). Another aspect which varied according to the consulted work is the positivity criterion.…”