Real-Time PCR Assay Compared to Nested PCR and Antigenemia Assays for Detecting Cytomegalovirus Reactivation in Adult T-Cell Leukemia-Lymphoma Patients

Ikewaki, Junji; Ohtsuka, Eiichi; Kawano, Rie; Ogata, Masao; Kojima, Hiroshi; Nasu, Masaru

doi:10.1128/jcm.41.9.4382-4387.2003

Cited by 25 publications

(13 citation statements)

References 26 publications

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“…These observations suggest that quantitative CMV PCR might be a useful tool to monitor the efficacy of anti-CMV therapy in bone marrow recipients and the discontinuation of CMV preemptive therapy based on quantitative CMV PCR might prevent a recurrent CMV disease. Ikewaki et al [48] observed that real-time PCR was more suitable for monitoring CMV reactivation in adult T-cell leukemia-lymphoma patients than the antigenemia assay. Ksouri et al [33] suggests that the CMV-DNA assay is better assay for monitoring patients receiving preemptive therapy, especially in CMV-GI disease, and that after CMV-infected cell destruction, the genome of defective virus could still be remaining and detected by PCR.…”

Section: Discussionmentioning

confidence: 98%

Monitoring of Cytomegalovirus Reactivation in Bone Marrow Transplant Recipients by Real-time PCR

Ghaffari

Obeidi

Dehghan

et al. 2008

Pathol. Oncol. Res.

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Cytomegalovirus (CMV) has been recognized as the most important viral pathogen in persons undergoing bone marrow transplantation (BMT). The aim was to develop a quantitative PCR assay to quantify CMV DNA in peripheral blood leukocytes (PBLs) of bone marrow transplantation (BMT) patients. An in-house real-time PCR assay based on TaqMan technology was developed to monitor the quantity of CMV DNA in PBLs of the BMT recipients. Sequential blood samples (415 specimens) were collected from 43 patients as weekly intervals until day 100 after transplantation. The CMV DNA was quantified in parallel with the pp65 antigenemia assay in PBL samples. Viral reactivation occurred in 51% and 41.8% of the recipients as detected by RQ-PCR and antigenemia assays respectively. There was a significant correlation between both assays (P < 0.0001); however, the RQ-PCR was more sensitive than the antigenemia. CMV DNA was detected by the RQ-PCR by a median of 14 days earlier than the antigenemia. Preemptive therapy was implemented in the antigenemia positive cases. The administration of ganciclovir led to a rapid decrease in the viral load. After preemptive therapy, the antigenemia achieved a negative result earlier than the RQ-PCR assay (a median of 17.5 days). An increase of viral load in both quantitative assays and of cyclosporine serum level were identified as the most significant risk factors for CMV reactivation. The quantitative CMV PCR might be a useful tool for monitoring the CMV reactivation and guiding the efficacy of the CMV preemptive therapy in BMT recipients.

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Section: Discussionmentioning

confidence: 98%

Monitoring of Cytomegalovirus Reactivation in Bone Marrow Transplant Recipients by Real-time PCR

Ghaffari

Obeidi

Dehghan

et al. 2008

Pathol. Oncol. Res.

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“…14 Peripheral blood leukocytes were isolated by dextran sedimentation, and leukocytes were cytocentrifuged on glass slides. The slides were air-dried, fixed with acetone, and then stained with peroxidase-conjugated monoclonal antibody HRP-C7, which specifically binds the pp65 antigen of CMV.…”

Section: Antigenemia Assaymentioning

confidence: 99%

Real-time PCR assays based on distinct genomic regions for cytomegalovirus reactivation following hematopoietic stem cell transplantation

Ikewaki

Ohtsuka

Satou

et al. 2004

Bone Marrow Transplant

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Summary:Real-time PCR has many advantages compared with antigenemia and qualitative PCR assays for detecting cytomegalovirus (CMV) infection in patients following SCT. However, the procedure used in each report was not standardized. This study compares the CMV load detected by real-time PCR assays amplifying distinct genomic regions. Real-time PCR assays based on US17, UL65, immediate early protein (IE) and glycoprotein B(gB) were selected and comparisons were made between each genomic region, and with antigenemia and nested PCR (IE region) in 18 SCT patients. The CMV load detected by real-time PCR using all combinations of primers targeting distinct genomic regions and by antigenemia assays correlated well. However, US17 and UL65-PCR could detect CMV earlier than gB-PCR, antigenemia and nested PCR assays. In longitudinal analysis, gB-PCR demonstrated a trend for showing a lower viral load in some patients than US17-, UL65-and IE-PCR. Moreover, the results suggest that a cutoff level of 500 copies/ml might be used to decide when to initiate treatment. We propose that monitoring should be carried out using real-time PCR assays targeting the US17 region and that a CMV load of 500 copies/ml could be used as a cutoff value for initiating treatment in patients following SCT, receiving immunoglobulin prophylaxis.

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“…Reports of the change in CMV antigenemia during GCV treatment in ATL are scarce [4]. Especially, there are no reports which mentioned the relationship between CMV infection, CMV antigenemia, treatment for CMV infection, and the clinical course of ATL.…”

mentioning

confidence: 98%

Maintenance and preemptive therapy with ganciclovir for cytomegalovirus colitis with extremely high antigenemia in adult T-cell leukemia

et al. 2009

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Patients with adult T-cell leukemia (ATL) are susceptible to opportunistic infections. The major cause of death in ATL patients is cytomegalovirus (CMV) pneumonia [1]. However, the onset of CMV infection in a very early phase of ATL is uncommon [2]. In addition, there are no detailed descriptions concerning the effect of oral ganciclovir (GCV) in ATL. Herein, we report a case with ATL who demonstrated CMV colitis on the second day of prednisolone administration.A 48-year-old female was admitted because of fever, erythema, and fatigue. Her white blood cell (WBC) count was 18,400/mm 3 (atypical lymphocytes including flower cells: 48.5%). The left and right cervical, axillary, and right inguinal lymph nodes were swollen. Erythema of the face, limbs, and neck was observed. Histopathologic findings of biopsy specimens from erythema sites demonstrated the possibility of ATL cells having invaded the skin. The lactate dehydrogenase (LDH) score was 505 U/L (more than twice the normal upper limit) and human T lymphotrophic virus type 1 provirus was detected on Southern blot analysis. Therefore, we diagnosed this case as acutetype ATL. The compensated calcium level was normal (9.6 mg/dL), and there were no other sites of involvement such as the liver, spleen, and cerebrospinal fluid. She was started on prednisolone for ATL. On the day following the start of administration, diarrhea and abdominal pain developed. We started chemotherapy primarily according to the LSG 15 protocol [3], because it was possible at that time that her diarrhea was caused by ATL cell invasion of the colon. However, an abdominal computed tomography scan showed ascites and mucosal thickening of the colon, which indicated colitis. Histopathologic findings of biopsy specimens from edematous lesions of the rectum detected by proctoscopy demonstrated intranuclear CMV inclusion bodies in endothelial cells but not the invasion of ATL cells. In addition, there were 3411 and 3258 CMV antigenpositive leukocytes each per 15 9 10 4 WBCs, and other pathogens which can induce diarrhea were not detected. No other manifestations induced by CMV infection such as CMV pneumonia and retinitis appeared. Therefore, the diagnosis of CMV colitis was confirmed. She received intravenous GCV. Diarrhea improved and the score of CMV antigenemia gradually declined (Fig. 1). After the diarrhea had fully resolved and informed consent was given, we continued maintenance therapy with oral GCV until CMV antigenemia became negative. Thereafter, we chose preemptive therapy involving oral GCV along with the monitoring of CMV antigenemia. This patient received allogeneic bone marrow transplantation (BMT) four months after the diagnosis of CMV colitis. We continued the same preemptive therapy until BMT. CMV infection never recurred before BMT. In addition, chemotherapy was effective. After chemotherapy, atypical lymphocytes in the peripheral blood disappeared, and the score of the soluble interleukin-2 receptor was reduced (Fig. 1). Swollen lymph nodes regressed. In addition, the level of ...

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Real-Time PCR Assay Compared to Nested PCR and Antigenemia Assays for Detecting Cytomegalovirus Reactivation in Adult T-Cell Leukemia-Lymphoma Patients

Cited by 25 publications

References 26 publications

Monitoring of Cytomegalovirus Reactivation in Bone Marrow Transplant Recipients by Real-time PCR

Monitoring of Cytomegalovirus Reactivation in Bone Marrow Transplant Recipients by Real-time PCR

Real-time PCR assays based on distinct genomic regions for cytomegalovirus reactivation following hematopoietic stem cell transplantation

Maintenance and preemptive therapy with ganciclovir for cytomegalovirus colitis with extremely high antigenemia in adult T-cell leukemia

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