A prospective study of a quantitative PCR ELISA assay for the diagnosis of CMV pneumonia in lung and heart-transplant recipients

Barber, Lisa G.; Egan, Jim; Lomax, Joyce; Haider, Yussef; Yonan, Nizar; Woodcock, Ashley; Turner, Andrew

doi:10.1016/s1053-2498(00)00136-4

Cited by 13 publications

(9 citation statements)

References 27 publications

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“…The measurement of viral load by quantitative PCR correlates with the evolution of CMV infection. Several studies have demonstrated a positive relationship between high CMV DNA levels in blood and the clinical progression of CMV infection either in solid organ or in bone marrow transplantation [Roberts et al, 1998;Vanpoucke et al, 1999;Barber et al, 2000;Camargo et al, 2001;Guiver et al, 2001;Paya, 2001;Razonable et al, 2002a,b]. The study of CMV dynamics through the definition of time points and DNA thresholds during the follow-up is a promising way to identify patients at higher risk of CMV infection evolution and optimize pre-emptive treatment [Roberts et al, 1998;Sia and Patel, 2000c;Ghisetti et al, 2004;Mattes et al, 2005].…”

Section: Introductionmentioning

confidence: 99%

Quantitation of cytomegalovirus DNA by real-time polymerase chain reaction in peripheral blood specimens of patients with solid organ transplants: Comparison with end-point PCR and pp65 antigen test

et al. 2006

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The polymerase chain reaction (PCR) for cytomegalovirus (CMV) DNA quantitation provides sensitive and specific data for detecting CMV as well as monitoring the infection and determining the appropriate antiviral strategy. A recently introduced real-time PCR assay for CMV DNA quantitation was applied on 158 peripheral blood leukocytes (PBLs) from 32 liver-transplanted patients with CMV asymptomatic infection and correlated with a commercial quantitative end-point PCR (COBAS AMPLICOR CMV Monitor) and CMV pp65 antigenemia. A good correlation was found between real-time PCR and pp65 antigen test (r2 = 0.691) and the two PCR assays (r2 = 0.761). Real-time PCR data were higher in pre-emptive treated patients (>20 pp65 + positive cells, median CMV DNA value: 3.8 log(10) copies/500,000 PBLs) than in not-treated ones (2.9 logs). According to pp65 levels of 0, 1-10, 11-20, 21-50, 51-100, and >100 positive cells/200,000 PBLs, median CMV DNA by real-time PCR was 2.6, 3.0, 3.6, 4.0, 4.2, and 4.8 logs, respectively, (CMV DNA levels by COBAS AMPLICOR: 2.8, 2.9, 3.8, 3.7, 3.9, and 4.0 logs). For samples with >20 pp65 + cells, real-time PCR gave significantly higher values than in groups with <20 pp65 + cells, whereas the COBAS AMPLICOR results showed a slower progression rate. Dilutions of CMV AD169 strain were used to probe real-time PCR reproducibility (between and intra-assay variability <2%) and sensitivity (100% detection rate at 10 copies/reaction, 28.5% with end-point PCR). In conclusion, real-time PCR significantly improves the study of CMV DNA dynamics due to a more reliable quantitation of CMV DNA for moderate and high DNA level compared to end-point PCR with better sensitivity and specificity. Real-time PCR provides more precise information for evaluating infection progress and assessing antiviral response, simplifying and accelerating the process of producing a reliable quantitation of CMV DNA for clinical purposes.

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Section: Introductionmentioning

confidence: 99%

Quantitation of cytomegalovirus DNA by real-time polymerase chain reaction in peripheral blood specimens of patients with solid organ transplants: Comparison with end-point PCR and pp65 antigen test

et al. 2006

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“…Several studies have shown a positive relationship between high CMV DNA levels in blood and the clinical progression of CMV infection either in solid organ or in bone marrow transplantation [Roberts et al, 1998;Vanpoucke et al, 1999;Barber et al, 2000;Paya et al, 2000;Sia et al, 2000a;Camargo et al, 2001;Guiver et al, 2001;Michaelides et al, 2001;Rollag et al, 2002;Razonable et al, 2002b]. The use of quantitative PCR data through the definition of time points and CMV DNA thresholds during the recipient follow-up provides an early detection of patients at high risk of CMV disease.…”

Section: Introductionmentioning

confidence: 99%

Quantitation of cytomegalovirus DNA by the polymerase chain reaction as a predictor of disease in solid organ transplantation

Ghisetti

Barbui

Franchello

et al. 2004

Journal of Medical Virology

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Cytomegalovirus (CMV) infection is an important cause of morbidity in solid organ recipients. Early markers to identify the progress of the infection and patients at high risk are required in order to apply a strategy of pre-emptive therapy. The efficacy of pre-emptive therapy relies on accurate laboratory tests to monitor CMV infection. The evaluation of CMV DNA kinetics by the polymerase chain reaction (PCR) is widely used for the management of CMV infection but markers predicting the progression of the infection and standardization of the technique are essential for the clinical interpretation of PCR results. A commercially available PCR system, the COBAS AMPLICOR Monitor (Roche Diagnostics, Brachburg, NJ), was used for the quantitation of CMV DNA in weekly blood samples (n = 504) from 47 solid organ recipients in the first 6 months after transplantation. PCR results were evaluated according to the development of clinical disease in order to find a DNA threshold and time points predicting the progression of CMV infection. Week 4 from transplantation was the earliest time point to note a significant difference between those patients who eventually developed CMV disease (n = 30) and those who remained asymptomatically infected (n = 17). At week 4, viral loads were significantly higher in patients who developed CMV disease than in asymptomatic infections (median value: 4 log(10)/10(6) leukocytes vs. 2.8, P < 0.0001). At week 4, a DNA level >/=4 log(10)/10(6) leukocytes was associated with a 45.37 odds ratio for CMV disease. Any increase >/=1 log from the first DNA detection to week 4 correlated with the clinical progression of CMV infection (odds ratio 1.74). In those patients who were treated with anti-CMV therapy, a >97% reduction of the baseline viral load was associated with a complete therapeutic success. In conclusion, CMV infection is a highly dynamic process and the quantitation of CMV DNA by PCR is a powerful marker to control successfully the infection, but a strict follow-up of the recipient and standardized PCR tests are mandatory for the best management of the infection.

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“…Polymerase chain reaction (PCR) techniques and hybrid capture assays can detect and quantify very small amounts of CMV DNA in clinical specimens. The clinical role of these approaches is rapidly evolving (36,37). Viral DNA can be quantified in different blood compartments (leukocytes, plasma, or whole blood), but several studies have shown that whole blood is the specimen of choice for CMV DNAemia quantification, since it allows determination of both cell-free and cell-associated virus (38)(39)(40)(41).…”

Section: Viral Dna Quantificationmentioning

confidence: 99%

CMV management in lung transplant recipients

Solidoro¹,

Costa²,

Libertucci³

et al. 2014

sob

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recipients management relative to the mortality and morbidity and, on the other hand, to the availability of effective antiviral agents as well as new diagnostic techniques and the prophylaxis, diagnosis, and treatment of CMV infections. In lung transplant recipients management is evolving with experience and we are living the sunrise of a new personalized or tailored management era matching, almost in real time, infection, diagnosis, immunity status and therapy. KEY WORDS: CMV infection, lung transplant, acute rejection, chronic rejection, hyperimmune globulins. IntroductionCytomegalovirus (CMV), a member of the beta herpes virus group, causes a spectrum of disorders, which includes serious illness in organ transplant recipients (1-4). Its presence in biological specimens and its role in co-infections and rejection is far to be fully defined. It is the second most common infection among lung transplant recipients, after bacterial pneumonia (5, 6), and is associated with significant morbidity and mortality after transplantation. Although the availability of effective antiviral drugs has decreased CMV-related mortality, the attention in last years has been addressed to its immunobiological role: in fact CMV infections or pneumonia have been associated to chronic rejection (bronchiolitis obliterans syndrome, BOS) in some, but not all, studies (7,8). The endothelium represents the anatomical and functional interface between engrafted donor tissue and the host immune system and interacts dynamically in immunomodulation. This implies an important role for the endothelial cell (EC) in the initiation, maintenance and/or termination of immune interactions affecting the fate of allograft (9). CMV does not directly induce HLA class II on infected human EC, but the rejection associated class II induction within the allograft might be a consequence of IFN-gamma release by activated T CD4+ cells in response to CMV infection (10).

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A prospective study of a quantitative PCR ELISA assay for the diagnosis of CMV pneumonia in lung and heart-transplant recipients

Cited by 13 publications

References 27 publications

Quantitation of cytomegalovirus DNA by real-time polymerase chain reaction in peripheral blood specimens of patients with solid organ transplants: Comparison with end-point PCR and pp65 antigen test

Quantitation of cytomegalovirus DNA by real-time polymerase chain reaction in peripheral blood specimens of patients with solid organ transplants: Comparison with end-point PCR and pp65 antigen test

Quantitation of cytomegalovirus DNA by the polymerase chain reaction as a predictor of disease in solid organ transplantation

CMV management in lung transplant recipients

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