Başkent Üniversitesi Makaleler

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Subject: search.filters.subject.Human cytomegalovirus

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    Quantification of Human Cytomegalovirus DNA by a New Capture Hybrid Polymerase Chain Reaction Enzyme-Linked Immunosorbent Assay in Plasma and Peripheral Blood Mononuclear Cells of Bone Marrow Transplant Recipients
    (Başkent Üniversitesi, 2008-12) Ziyaeyan, Mazyar; Kadivar, Mohammad; Pourabbas, Bahman; Mahboudi, Fereidoun; Ramzi, Mani; Alborzi, Abdolvahab; Sabahi, Farzaneh
    Objectives: Quantitative monitoring of human cytomegalovirus infections is helpful in determining appropriate antiviral management in patients who receive bone marrow transplants. We sought to design and evaluate a new cytomegalovirus capture hybrid polymerase chain reaction enzyme-linked immunosorbent assay (PCR-ELISA) in plasma and peripheral blood mononuclear cells to monitor cytomegalovirus infection in bone marrow transplant recipients. Patients and Methods: Twenty-six patients who received allogeneic bone marrow transplants, including 17 male patients and 9 female patients (9 adults, 17 children), were enrolled in this study. A total of 313 consecutive whole blood specimens, before and from 7 to 120 days after transplant, was evaluated in the study. A newly designed biotinylated probe-mediated quantitative competitive PCR-ELISA test was used to determine cyto­megalovirus load in specimens of peripheral blood mononuclear cells and plasma. Results: All 26 patients were cytomegalovirus seropositive before transplant. Capture hybrid PCR-ELISA of peripheral blood mononuclear cells detected cytomegalovirus DNA in 287 of 313 specimens (91.7%) even in cases with no active cytomegalovirus infection. in plasma, cyto­megalovirus DNA was detected in 114 of 313 specimens (36.4%). Increasing titers of cyto­megalovirus DNA were detected in 14 of 26 patients (53.8%). Conclusions: The quantitative capture hybrid PCR-ELISA was able to diagnose and monitor cytomegalovirus infection in patients who received bone marrow transplants. Detection of cyto­megalovirus DNA in plasma was more predictive of the onset of cytomegalovirus-related clinical symptoms, compared to detection in peripheral blood mononuclear cells.
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    Comparison of Antigenemia Assay and Semiquantitative Polymerase Chain Reaction Test For Monitoring Active Cytomegalovirus Infection in Allogeneic Hematopoietic Cell Transplant Recipients
    (Başkent Üniversitesi, 2008-06) Pajand, O.; Abdossamadi, Z.; Bahador, A.; Hojabri, Z.; Fatolahzadeh, B.; Mousavi, S. A.; Ziyaeyan, M.
    Objectives: We sought to compare the antigenemia assay and in-house semiquantitative polymerase chain reaction to monitor human cytomegalovirus infection after transplant in hematopoietic cell transplant recipients. Materials and Methods: A pp65 antigen test for poly­morphonuclear leukocytes and a semi­quantitative polymerase chain reaction for whole blood were performed for 201 samples obtained from 26 hematopoietic cell transplant recipients over a 3-month surveillance period. Results: Fourteen episodes of antigenemia positivity were detected in 7 patients in whom human cytomegalovirus DNA loads and pp65-positive cells ranged between < 102 to 2.96 × 104 copies/mL and 0-35/ 5 × 104 polymorphonuclear leukocytes, respectively. A significant correlation was detected between human cytomegalovirus DNA load and the antigenemia test. A receiver operating characteristic analysis determined 5000 copies/mL of human cytomegalovirus as the threshold value for initiation of ganciclovir therapy. Conclusions: Based on a comparison of the pp65 antigenemia assay, quantification of human cytomegalovirus DNA in whole blood can be used to guide clinical management of hematopoietic cell transplant recipients. This approach may have important advantages including superior sensitivity and efficient monitoring of preemptive therapy, allowing inclusion of kinetic criteria in clinical guidelines. Furthermore, a high human cyto­megalovirus load among patients with grade II-IV acute graft-versus-host disease may indicate a high risk of human cytomegalovirus disease among hematopoietic cell transplant patients. Human cytomegalovirus reactivation must be monitored using more-sensitive assays such as real-time polymerase chain reaction.
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    Monitoring Human Cytomegalovirus (HCMV) in HCMV-Seropositive Orthotopic Liver-transplant Recipients by Means of Quantitative Real-time Polymerase Chain Reaction
    (Başkent Üniversitesi, 2006-12) Mengelle, Catherine; Abravanel-Legrand, Florence; Kamar, Nassim; Alain, Sophie; Basse, Grégoire; Pillet, Adèle; Lavayssière, Laurence; Suc, Bertrand; Izopet, Jacques; Rostaing, Lionel
    Objective: Human Cytomegalovirus can be reactivated after orthotopic liver transplantation in patients who are seropositive for cytomegalovirus. Whether those cytomegalovirus-seropositive patients require immediate posttransplant (anti)cytomegalovirus prophylactic therapy or preemptive treatment as opposed to deferred treatment remains controversial. The aims of our study were to evaluate the relevance of cytomegalovirus monitoring with quantitative real-time polymerase chain reaction in whole blood and to analyze the factors that determine the treatment of the first episode of cytomegalovirus infection with intravenous ganciclovir in seropositive liver-transplant patients. Patients and Methods: Forty-two cytomegalovirus-seropositive liver-transplant patients were assessed for cytomegalovirus DNAemia every 2 weeks until posttransplant day 90 and every 3 to 4 weeks until day 180. Biochemical and hematologic parameters were also prospectively monitored. Results: Cytomegalovirus DNAemia was detected at least once in 27 patients (64%). Treatment was initiated in 12 patients (group 1) but not in 15 others (group 2). Median HCMV viral loads of the first positive and the highest DNAemia were statistically higher in group 1 than in group 2 (P = 0.01). Univariate analysis of DNAemia showed that alkaline phosphatase levels were significantly higher in group 1 than in group 2 (P = .0011) and that hemoglobin levels were significantly lower in group 1 than in group 2 (P = .0443). The results of multivariate analysis showed that the only factor that predicted the treatment of the first episode of HCMV DNAemia was a level of alkaline phosphatase greater than 150 IU/L [odds ratio, 20; range, 1.97-203.32; P = .01]. Conclusions: A combination of criteria, including viral-load kinetics, clinical factors, alkaline phosphatase levels (in particular), and the patient’s immune condition, is required to efficiently monitor patients who are seropositive for cytomegalovirus after orthotopic liver transplantation.
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    Diagnosis and Monitoring of Human Cytomegalovirus Infection in Bone Marrow Transplant Recipients by Quantitative Competitive PCR
    (Başkent Üniversitesi, 2006-06) Ziyaeyan, Mazyar; Sabahi, Farzaneh; Alborzi, Abdolvahab; Mahboudi, Fereidoun; Kazemnejad, Anooshirvan; Ramzi, Mani; Moravej, Ali; Jaberi, Marjan Mojtahed
    Objectives: Human cytomegalovirus (HCMV) is a common cause of infection worldwide. Severe cytomegalovirus disease is usually observed in immunodeficient individuals such as bone marrow transplant (BMT) or AIDS patients. In these patients, proof of viral presence is not enough for making clinical decisions; one must report the quantity of virus or viral load in appropriate clinical specimens to demonstrate the relationship between disease severity and HCMV infection. The goal of this study was to use quantitative competitive polymerase chain reaction (PCR) to determine HCMV viral load in 26 BMT recipients. Materials and Methods: Peripheral blood was collected weekly for 100 days from 26 BMT recipients. Qualitative and quantitative competitive PCRs on 105 mononuclear cells were performed for each patient. The same tests were performed once for each of 26 donors. In addition, the anti-HCMV humoral response was detected by performing IgM and IgG ELISAs in donors and recipients prior to transplantation. Results: Of 26 BMT donors and recipients, 25 and 26 were IgG positive, and 2 and 6 had HCMV-specific IgM antibodies, respectively. From 313 total clinical specimens tested, 255 had positive qualitative PCR results. Results of quantitative PCR on the same specimens demonstrated that in 14 patients, viral copy number per 105 cells had increased, pointing toward HCMV reactivation. In others, changes in viral copy number were mostly around 100/105 cells, with an upper limit of 300/105 cells. Conclusions: Owing to the high prevalence of cytomegalovirus in our country, the chance of viral reactivation and HCMV infection/disease upon transplantation must be seriously considered. Therefore, use of quantitative PCR in PCR-positive patients is highly recommended to demonstrate active infection that may lead to HCMV disease during the posttransplant period. This also could help physicians begin pre-emptive therapy that would be for a shorter treatment period and provide for better outcomes in infected BMT patients.
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    Qualitative Detection of Human Cytomegalovirus DNA in the Plasma of Bone Marrow Transplant Recipients: Value as a Predictor of Disease Progression
    (Başkent Üniversitesi, 2004-06) Behzad-Behbahani, A; Rasoli, M; Ramzi, M; Nourani, H; Alborzi, A; Ehsanipour, F
    Objective: The aim of this prospective study was to determine whether human cytomegalovirus (HCMV)-DNA detected by polymerase chain reaction (PCR) analysis in the plasma of bone marrow transplant (BMT) patients is a predictor of HCMV disease progression. Methods: Plasma samples were collected from 15 patients who received allogenic BMTs. Each individual was sampled 1 week before and then weekly for 17 weeks after transplantation. The 270 plasma specimens were processed with a PCR method for detecting HCMV-DNA. Patients were also physically examined for signs or symptoms of HCMV-related disease. Results: Eight (53.5%) of the 15 patients tested positive for HCMV-DNA. Two (25%) of these 8 individuals also had positive PCR findings before transplantation. Six (75%) of the 8 HCMV-DNA-positive patients had positive plasma-PCR results a week before clinical symptoms developed. The other 2 (25%) remained asymptomatic throughout their hospital stay. All 6 symptomatic cases were treated with ganciclovir, and 4 converted to negative plasma-PCR status at a median of 21 days. There was a significant correlation between PCR-detection of HCMVDNA in plasma and presence of HCMV-related symptoms (P < 0.01). Conclusion: Qualitative plasma-PCR analysis before and after bone marrow transplantation is a valuable way to screen for HCMV infection in BMT patients. Plasma-PCR monitoring of HCMV activity in this patient group might make it possible to administer an antiviral drug and thus reduce mortality. However, quantitative PCR is still considered the best way to accurately identify active HCMV infection and monitor treatment.