New Molecular Assays to Predict Occurrence of Cytomegalovirus Disease in Renal Transplant Recipients

Pellegrin, Isabelle; Garrigue, Isabelle; Ekouévi, Didier K.; Couzi, Lionel; Merville, Pierre; Merel, Patrick; Chêne, Geneviève; Schrive, Marie-Hélène; Trimoulet, Pascale; Lafon, Marie‐Edith; Fleury, Hervé

doi:10.1086/315688

Cited by 41 publications

(36 citation statements)

References 36 publications

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“…The use of a more sensitive recombinant antigen ELISA IgM assay (similar to the one used in our study), was found to have some clinical utility in predicting CMV disease in one study of 40 liver transplant recipients, although again DNAemia generally preceded detection of antibody (15). Based on these data, in the past several years, molecular detection methods have generally replaced serology and culture based tests for the diagnosis of active CMV disease (16)(17)(18). There are, however, no previous studies assessing the utility of CMV serology testing in a large cohort of D+/R− patients receiving antiviral prophylaxis, for the purposes of identifying patients at risk of late-onset CMV disease.…”

Section: Discussionmentioning

confidence: 62%

Clinical Utility of Cytomegalovirus (CMV) Serology Testing in High-risk CMV D+/R- Transplant Recipients

Humar

Mazzulli

Moussa

et al. 2005

American Journal of Transplantation

147

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Late-onset cytomegalovirus (CMV) disease is a significant problem in D+/R− solid organ transplant (SOT)patients who receive antiviral prophylaxis. We assessed the clinical utility of CMV IgG and IgM serology testing for predicting late-onset CMV disease. We evaluated 352 D+/R− transplant recipients who participated in a trial comparing 100 days of ganciclovir versus valganciclovir prophylaxis. CMV serology was assessed on day 28, 56, 100, and 6 and 12 months post-transplant. IgG seroconversion occurred in 26.9% of patients by day 100, and in 63.4% and 75.3% by 6 and 12 months, respectively. IgM seroconversion occurred in 8.3%, 41.8% and 54.9% by day 100, month 6 and month 12, respectively. Seroconversion by day 100 (end of prophylaxis) was not predictive of subsequent CMV disease (CMV disease 13.3% if seropositive vs. 17.8% if seronegative; p = NS). However, at 6 months post-transplant, IgG serostatus was predictive of subsequent CMV disease between month 6 and 12 (CMV disease 1.3% if seropositive vs. 10.0% if seronegative; p = 0.002). In D+/R− patients, CMV serology testing is for the most part not clinically useful for predicting subsequent disease. However, seroconversion by 6 months may be useful for identifying patients at risk of late-onset CMV disease.

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Section: Discussionmentioning

confidence: 62%

Clinical Utility of Cytomegalovirus (CMV) Serology Testing in High-risk CMV D+/R- Transplant Recipients

Humar

Mazzulli

Moussa

et al. 2005

American Journal of Transplantation

147

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“…The NucliSens assay (Organon Teknika Diagnostics, Boxtel, The Netherlands), an isothermal nucleic acid amplification reaction assay, detects the presence of CMV latemRNA pp67. The presence of mRNA pp67 indicates active viral replication, and its detection is a marker for active CMV infection (18,19). Nevertheless, this assay has less sensitivity than DNA amplification and antigenemia assays for detection of CMV infection.…”

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confidence: 99%

“…The lower sensitivity of the assay may result in failure to detect or predict CMV disease in all patients (22). In one study, the assay did not detect the mRNA transcripts in 4 of 11 patients who developed CMV disease (19). A commercially available COBAS Amplicor CMV Monitor (CACM) assay has been developed by Roche Diagnostics (Branchburg, N.J.) for the determination of CMV DNA load (5,6,30).…”

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confidence: 99%

Comparison of LightCycler-Based PCR, COBAS Amplicor CMV Monitor, and pp65 Antigenemia Assays for Quantitative Measurement of Cytomegalovirus Viral Load in Peripheral Blood Specimens from Patients after Solid Organ Transplantation

et al. 2003

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In order to evaluate the LightCycler-based PCR (LC-PCR) as a diagnostic assay technique, a classical pp65antigenemia assay and the commercially available COBAS Amplicor CMV Monitor (CACM) assay were compared to the LC-PCR assay for the detection and quantitation of cytomegalovirus (CMV) load in 404 parallel specimens of peripheral blood from 66 patients after solid organ transplantation. A good correlation existed among these three assays (r Х 0.6, P < 0.0001). The LC-PCR assay was the most sensitive (54% of specimens positive) compared to the CACM (48.6%) and the pp65 antigenemia (26%) assays. The LC-PCR assay detected all samples found positive by using both the CMV pp65 antigenemia assay and the CACM assay. The LC-PCR also had the widest dynamic range (from 250 to 10 7 DNA copies/ml of plasma). No cross-reactions were found among CMV and Epstein-Barr virus, varicella-zoster virus, or herpes simplex virus in the LC-PCR by using amplification with specifically designed primer pairs. Precision, expressed as the coefficient of variation, was <3% with standard DNA from cell cultures and between 6.55 and 14.1% with clinical specimens in repeat LC-PCR runs. One run of the LC-PCR took half of the time required for the semiautomated CACM procedure. Because of its sensitivity, specificity, cost-effectiveness, and simplicity, the LC-PCR assay could replace the pp65 antigenemia and the CACM assays as the preferred technique for the surveillance, diagnosis, and monitoring of response of CMV diseases in high-risk populations.

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“…For the benefit of such strategies, quantitative detection of CMV DNA in the blood compartment by PCR is increasingly used to identify patients at risk for CMV disease. In addition, measurements of viral DNA load may be important for monitoring the efficacy of antiviral treatment and predicting the development of drug resistance (3,22,27).The genome of human CMV consists of a large (about 230,000-bp) double-stranded linear DNA molecule which is encapsidated within a double protein shell and a lipid envelope (24). Most of the CMV DNA in the blood compartment is present in abortively infected polymorphonuclear leukocytes (4,12,16,26); less DNA is found in peripheral blood mononuclear cells, part of which, upon differentiation into macrophages, support viral replication (12,26,28).…”

mentioning

confidence: 99%

mentioning

confidence: 99%

Human Cytomegalovirus DNA in Plasma and Serum Specimens of Renal Transplant Recipients Is Highly Fragmented

Boom¹,

Sol²,

Schuurman³

et al. 2002

J Clin Microbiol

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Quantitation of cytomegalovirus (CMV) DNA in plasma and serum by PCR is increasingly used to identify patients at risk for developing CMV disease and to monitor the efficacy of antiviral therapy. Although CMV DNA levels are generally interpreted as viral loads, the exact nature of the viral DNA in these specimens is unknown. We studied the state of CMV DNA in plasma and serum specimens obtained from three renal transplant recipients at peak viral DNA levels during primary CMV infection. For this purpose, DNA isolated from these specimens was fractionated by size, and CMV DNA levels in the resulting DNA fractions were measured by quantitative PCR targeted at large (578-bp) and small (134-bp) amplicons. These experiments showed that the molecular sizes of DNA fragments from which CMV DNA is amplified were small (<2,000 bp), indicating that CMV DNA in plasma and serum is highly fragmented. Furthermore, CMV DNA levels were consistently higher when targeted at the smaller amplicon, providing additional evidence for the fragmentation of viral DNA. In conclusion, the first results with three patients have shown that CMV DNA in plasma and serum is highly fragmented and does not necessarily reflect the amount of infectious virus. These observations have potential consequences for understanding CMV pathogenesis and interpreting CMV DNA levels in individual patient management.Infection with cytomegalovirus (CMV) (human herpesvirus 5) is an important cause of morbidity and mortality in immunocompromised individuals, such as transplant recipients and AIDS patients. In the management of CMV infection, preemptive treatment strategies, aimed at preventing CMV disease in high-risk patients, are receiving increasing attention. For the benefit of such strategies, quantitative detection of CMV DNA in the blood compartment by PCR is increasingly used to identify patients at risk for CMV disease. In addition, measurements of viral DNA load may be important for monitoring the efficacy of antiviral treatment and predicting the development of drug resistance (3,22,27).The genome of human CMV consists of a large (about 230,000-bp) double-stranded linear DNA molecule which is encapsidated within a double protein shell and a lipid envelope (24). Most of the CMV DNA in the blood compartment is present in abortively infected polymorphonuclear leukocytes (4,12,16,26); less DNA is found in peripheral blood mononuclear cells, part of which, upon differentiation into macrophages, support viral replication (12,26,28). In addition, circulating, productively infected endothelial cells may be a source of CMV DNA (16, 18). CMV DNA can also be detected in serum and plasma, which are convenient specimens for CMV DNA load measurement (10,15,19,29). In recent years, many studies on the qualitative and quantitative detection of CMV DNA in these specimens have been reported, which generally show that levels of CMV DNA found in plasma or serum are significantly lower than those found in white blood cells (4,5,13,15,33).At present, it is unclear whether CMV ...

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New Molecular Assays to Predict Occurrence of Cytomegalovirus Disease in Renal Transplant Recipients

Cited by 41 publications

References 36 publications

Clinical Utility of Cytomegalovirus (CMV) Serology Testing in High-risk CMV D+/R- Transplant Recipients

Clinical Utility of Cytomegalovirus (CMV) Serology Testing in High-risk CMV D+/R- Transplant Recipients

Comparison of LightCycler-Based PCR, COBAS Amplicor CMV Monitor, and pp65 Antigenemia Assays for Quantitative Measurement of Cytomegalovirus Viral Load in Peripheral Blood Specimens from Patients after Solid Organ Transplantation

Human Cytomegalovirus DNA in Plasma and Serum Specimens of Renal Transplant Recipients Is Highly Fragmented

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