Multicenter evaluation of PCR methods fordetecting CMV DNA in blood donors

Roback, John D.; Hillyer, Christopher D.; Drew, W. Lawrence; Laycock, Megan E.; Luka, János; Mocarski, Edward S.; Slobedman, Barry; Smith, James W.; Söderberg‐Nauclér, Cecilia; Todd, Deborah; Woxenius, Susanne; Busch, Michael P.

doi:10.1046/j.1537-2995.2001.41101249.x

Cited by 61 publications

(84 citation statements)

References 33 publications

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“…Indeed, we previously showed that 36% of HCMV IgG-negative blood donors with HCMV DNA-positive blood cells were IgG positive in an ELISA that uses a clinical isolate as antigen. 19 Similarly, in the present study, serum from 5 (42%) of 12 patients that were HCMV IgG negative according to three serology tests utilizing AD169-derived antigens, were IgG positive in an ELISA assay using antigens from a clinical isolate. Nevertheless, 9 (58%) of 12 patients were still HCMV IgG negative, although their tumors were positive for HCMV by immunohistochemistry with two different HCMV-specific antibodies, HCMV DNA were detected in their blood cells, and a majority of them had T cells that responded to HCMV peptides.…”

Section: E982391-4 Volume 4 Issue 2 Oncoimmunologysupporting

confidence: 72%

“…Such analyses have for obvious reasons not been done on tissues samples from healthy individuals, but a discrepancy between HCMV serology data and HCMV DNA detection in blood has been reported as discussed above. 17,18,19 Thus, several lines of evidence suggest that HCMV carriers are not always defined by serology, and our findings imply that this is frequently the case in GBM patients. We observed that 29% of GBM patients positive for HCMV DNA in blood cells and HCMV proteins in the tumor, were negative for HCMV IgG by serology, but 85% of them had T-cells that were reactive against HCMV IE and/or pp65 peptides.…”

Section: E982391-4 Volume 4 Issue 2 Oncoimmunologysupporting

confidence: 55%

“…We and others previously showed that commercial ELISA kits yield negative results for HCMV IgG in some patients who are positive for HCMV DNA. 17,18,19 We have also demonstrated that infectious virus could be reactivated from an HCMV DNA-positive but HCMV IgG-negative healthy blood donor 18 , and we recently also observed that a HCMV-seronegative mother whose breast milk was HCMV DNA positive transferred HCMV to her infant, who developed a HCMV infection (unpublished observation). Bianchi et al recently presented evidence that HCMV-DNA and proteins are present in brain tumors in the absence of detectable HCMV-specific antibodies against HCMV in the patients serum.…”

Section: Discussionmentioning

confidence: 77%

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Discordant humoral and cellular immune responses toCytomegalovirus(CMV) in glioblastoma patients whose tumors are positive for CMV

et al. 2015

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Submit your article to this journal Background. Glioblastoma (GBM) is the most common malignant brain tumor in adults and is nearly always fatal. Emerging evidence suggests that human Cytomegalovirus (HCMV) is present in 90-100% of GBMs and that add-on antiviral treatment for HCMV show promise to improve survival.Methods. In a randomized, placebo-controlled trial of valganciclovir in 42 GBM patients, blood samples were collected for analyses of HCMV DNA, RNA, reactivity against HCMV peptides, IgG, and IgM at baseline and at 3, 12, and 24 weeks of treatment.Results. All 42 tumors were positive for HCMV protein. All patients examined had at least one blood sample positive for HCMV DNA, 63% were HCMV RNA positive, and 21% were IgM positive. However, 29% of GBM patients were IgG negative for HCMV. Five of these samples were positive in an enzyme-linked immunosorbent assay (ELISA) that used antigens derived from a clinical isolate. Blood T cells from 11 of 13 (85%) HCMV IgG-negative GBM patients reacted against HCMV peptides. Valganciclovir did not affect IgG titers, DNA, or RNA levels of the HCMV immediate early (HCMV IE) gene in blood.Conclusion. In GBM patients, HCMV activity is higher than in healthy controls and serology is a poor test to define previous or active HCMV infection in these patients.

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Section: E982391-4 Volume 4 Issue 2 Oncoimmunologysupporting

confidence: 72%

Section: E982391-4 Volume 4 Issue 2 Oncoimmunologysupporting

confidence: 55%

Section: Discussionmentioning

confidence: 77%

See 1 more Smart Citation

Discordant humoral and cellular immune responses toCytomegalovirus(CMV) in glioblastoma patients whose tumors are positive for CMV

et al. 2015

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“…17 However, it should be noted that CMV genome have been detected in clinical samples of CMV-seronegative, healthy blood donors. 52 Larsson et al even detected the CMV genome in 55% of peripheral leukocytes of CMV-seronegative blood donors. 53 The presence of the CMV genome in samples of CMV-seronegative individuals, however, could not be confirmed by other large studies.…”

Section: Discussionmentioning

confidence: 99%

Recombinant antibodies encoded by IGHV1-69 react with pUL32, a phosphoprotein of cytomegalovirus and B-cell superantigen

Steininger

Widhopf²,

Ghia

et al. 2012

Blood

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IntroductionChronic lymphocytic leukemia (CLL) is a low-grade malignancy of mature B cells that express low levels of functional surface membrane immunoglobulin (Ig), the B-cell antigen receptor (BCR). 1 The Ig repertoire of healthy B cells is vast and in a constant state of flux. Diversity is generated in particular by recombination of germ line segments and, after ligation of antigen by a BCR with adequate affinity, somatic hypermutation. This process of Ig maturation is initiated on first encounter with a specific antigen, and subsequent or prolonged antigen exposure leads to generation of antibodies of even higher affinity to the specific antigen. 2 In contrast, the Ig repertoire expressed by leukemia B cells from unrelated CLL patients is highly restricted and skewed, and it is not representative of the Ig repertoire expressed by naive B cells. [3][4][5] Approximately 50% of CLL B cells express Ig heavy chains encoded by unmutated Ig heavy chain variable (IGHV) region genes, and of these more than 20% express closely homologous, if not identical, stereotyped heavy chain third complementarity determining regions (HCDR3). 5-7 IGHV1-69 genes belong to the most frequently used IGHV genes and are mostly unmutated. 6 Greater than 70% of IGHV1-69 cases use a 51p1-like allele and these 51p1-encoded Ig heavy chains are clearly distinct from those found in the normal adult blood B-cell repertoire. 8,9 The highly restricted Ig repertoire expressed by CLL B cells is the basis of the widely accepted notion that a limited set of antigens may react with the Ig of the leukemic cells and that this interaction is relevant for the homeostasis of leukemic cells.For CLL B cells, the functional consequences of Ig-mediated ligation of antigens are poorly defined, primarily because of the lack of knowledge of the relevant antigens. Therefore, a search for relevant antigens has been conducted in recent years. Candidate antigens should be distributed widely and provide chronic or repetitive stimuli for B cells as found in selected infections or autoimmune diseases. 10,11 CLL recombinant antibodies (rAbs) that were derived from Ig expressed by leukemic cells were generated by several research groups to screen for reactivity with antigen panels that represent antibody targets in autoimmune diseases for reactivity. This limited set of CLL rAbs were found to react with multiple different antigens, including IgG, cardiolipin, singlestranded DNA, actin, thyroglobulin, 12 nonmuscle myosin heavy chain IIA, 13 and molecular motifs on oxidized low-density lipoprotein and apoptotic cells. 14 Binding reactivity of several Abs with multiple antigens suggests the presence of a common antigenic determinant on the proteins screened. Alternatively, reactivity of multiple CLL rAbs with very different stereotyped Ab motifs could indicate the involvement of a superantigen that has antigen-receptormediated interactions with a large proportion of the leukemic cell pool. However, the identity and specificity of these antigenantibody interactions that pres...

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“…Therefore, the molecular detection of CMV DNA is essential for the diagnosis of acute CMV and the assessment of disease progression through monitoring of active CMV infections (virus clearance versus persistence). Early studies using different PCR assays, mainly inhouse tests, revealed considerable differences in their sensitivities and specificities, confirming the need for implementation of validated CMV assays (15). Therefore, the performance of different commercially available amplification systems was evaluated.…”

Section: Discussionmentioning

confidence: 99%

Systematic Evaluation of Different Nucleic Acid Amplification Assays for Cytomegalovirus Detection: Feasibility of Blood Donor Screening

2015

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Acute primary cytomegalovirus (CMV) infections, which commonly occur asymptomatically among blood donors, represent a significant risk for serious morbidity in immunocompromised patients (a major group of transfusion recipients). We implemented a routine CMV pool screening procedure for plasma for the identification of CMV DNA-positive donors, and we evaluated the sensitivities and performance of different CMV DNA amplification systems. Minipools (MPs) of samples from 18,405 individual donors (54,451 donations) were screened for CMV DNA using the RealStar CMV PCR assay (Altona Diagnostic Technologies), with a minimum detection limit of 11.14 IU/ml. DNA was extracted with a high-volume protocol (4.8 ml, Chemagic Viral 5K kit; PerkinElmer) for blood donor pool screening (MP-nucleic acid testing [NAT]) and with the Nuclisens easyMAG system (0. H uman cytomegalovirus (CMV) is a ubiquitous viral pathogen that causes mostly asymptomatic disease in immunocompetent individuals. In immunocompromised patients, however, CMV infection represents a significant risk for serious morbidity, e.g., due to interstitial pneumonia or hepatitis (1-3). Immunocompromised patients, such as patients undergoing hematopoietic stem cell transplantation (HSCT), solid-organ transplant recipients, infants with low birth weights, fetuses, pregnant woman, HIV patients, and patients being treated for hematological malignancies, belong to the major groups of transfusion recipients, and CMV-seronegative individuals were considered high-risk patients for transfusion-transmitted (TT)-CMV infections (4-6). The introduction of leukodepletion of blood products and provision of CMV-seronegative blood products reduced the incidence of TT-CMV infections in at-risk populations by 92%. However, TT-CMV breakthrough infections occur in 1 to 3% of high-risk patients who receive transfusions (4-6), possibly due to windowphase donations during acute primary CMV infections.The seroprevalence rates of CMV antibodies among blood donors show geographic differences, ranging from 45.8% in Germany to 96.5% in Brazil (3, 7). Primary CMV infections in blood donors occur in all age groups, with prevalence rates between 0.2 and 1.2% (3,6,8,9). The disease presentation is mostly asymptomatic, frequently with a prolonged course (10, 11). Mononucleosis-like symptoms are rare, whereas nonspecific viral disease symptoms occur often but not at significantly increased rates, compared to matched control groups (3,5).Determination of the prevalence of primary CMV infections among blood donors represents an important parameter for the effective prevention of 12). The aim of the present study was implementation of routine CMV DNA screening according to the setup used for our standard viral nucleic acid MATERIALS AND METHODSBlood donors. A total of 54,451 allogeneic blood donations from 18,405 individual German blood donors were routinely screened for the presence of CMV DNA by the Uni.Blutspendedienst OWL between March and June 2013. Master pools of 96 donations were set up by combin...

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Multicenter evaluation of PCR methods fordetecting CMV DNA in blood donors

Cited by 61 publications

References 33 publications

Discordant humoral and cellular immune responses toCytomegalovirus(CMV) in glioblastoma patients whose tumors are positive for CMV

Discordant humoral and cellular immune responses toCytomegalovirus(CMV) in glioblastoma patients whose tumors are positive for CMV

Recombinant antibodies encoded by IGHV1-69 react with pUL32, a phosphoprotein of cytomegalovirus and B-cell superantigen

Systematic Evaluation of Different Nucleic Acid Amplification Assays for Cytomegalovirus Detection: Feasibility of Blood Donor Screening

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