ArticleJournal of clinical microbiology1999
A highly sensitive assay for detection and quantitation of human cytomegalovirus DNA in serum and plasma by PCR and electrochemiluminescence.
Article in Journal of clinical microbiology, 1999. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 30 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
30 citing papers in PubMed.
- Epstein-Barr virus-targeted therapy in nasopharyngeal carcinoma.Journal of cancer research and clinical oncology · 2015Trial
- Shining a light on viral detection: a comparative study of electrochemical and electrochemiluminescence techniques for human cytomegalovirus.Analytical and bioanalytical chemistry · 2026Article
- Towards standardization in pig microbiome research based on a comprehensive twenty-year review.Animal microbiome · 2026Review
- A rapid purification method for trace amounts of cell-free DNA in urine.Analytical sciences : the international journal of the Japan Society for Analytical Chemistry · 2025Article
- IL-10-Secreting CD8Pathogens (Basel, Switzerland) · 2022Article
- Development of a novel anti-hepatitis B virus agent via Sp1.Scientific reports · 2020Article
- Assessing Anti-HCMV Cell Mediated Immune Responses in Transplant Recipients and Healthy Controls Using a Novel Functional Assay.Frontiers in cellular and infection microbiology · 2020Article
- Article
- High-resolution melting PCR assay, applicable for diagnostics and screening studies, allowing detection and differentiation of several Babesia spp. infecting humans and animals.Parasitology research · 2017Article
- The Contribution of Cytomegalovirus Infection to Immune Senescence Is Set by the Infectious Dose.Frontiers in immunology · 2017Article
- Advances in nanomaterials and their applications in point of care (POC) devices for the diagnosis of infectious diseases.Biotechnology advances · 2016Review
- Severe anemia in Malawian children.Malawi medical journal : the journal of Medical Association of Malawi · 2016Article
- Clonal evolution of CD8+ T cell responses against latent viruses: relationship among phenotype, localization, and function.Journal of virology · 2015Article
- Synthesis, labeling and bioanalytical applications of a tris(2,2'-bipyridyl)ruthenium(II)-based electrochemiluminescence probe.Nature protocols · 2014Article
- GS-8374, a prototype phosphonate-containing inhibitor of HIV-1 protease, effectively inhibits protease mutants with amino acid insertions.Journal of virology · 2014Article
- Performance evaluation of ExiStation HBV diagnostic system for hepatitis B virus DNA quantitation.Journal of virological methods · 2013Article
- Deep sequencing of antiviral T-cell responses to HCMV and EBV in humans reveals a stable repertoire that is maintained for many years.PLoS pathogens · 2012Article
- Rapid T cell repopulation after rabbit anti-thymocyte globulin (rATG) treatment is driven mainly by cytomegalovirus.Clinical and experimental immunology · 2012Article
- Cytomegalovirus-induced effector T cells cause endothelial cell damage.Clinical and vaccine immunology : CVI · 2012Article
- Persistent detection of varicella-zoster virus DNA in a previously healthy child after severe chickenpox.Journal of clinical microbiology · 2005Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We describe a diagnostic PCR assay (D-PCR) and a quantitative PCR assay (Q-PCR) for the detection of human cytomegalovirus (CMV) in plasma and serum. In the D-PCR, DNA was purified from plasma or serum together with internal control (IC) DNA, which monitored both DNA extraction efficiency and PCR efficiency. DNA was subjected to PCR with a single primer pair, and the amount of PCR products was determined by electrochemiluminescence (ECL) in the QPCR System 5000 (Perkin-Elmer) after hybridization with Tris (2,2'-bipyridine) ruthenium (II) chelate-labeled probes. The lower limit of sensitivity of the D-PCR was reached at about 25 CMV particles/ml. Even with extremely low DNA inputs (four molecules of IC DNA/200 microl of plasma), very high yields (near 100%) were reached. DNA extracted from specimens that were CMV positive by the D-PCR was subsequently used in the Q-PCR, which was similar to the D-PCR. The viral load was calculated directly from the ratio of CMV and IC signals obtained by ECL. The Q-PCR assay is quantitative in the range of 100 to 150,000 copies of CMV/ml, independent of the anticoagulant. Interassay variation, intra-assay variation, and interspecimen variation were about 25%, suggesting that the Q-PCR will reliably detect fourfold differences in viral load. Comparison of paired serum and plasma specimens from CMV-infected individuals showed that serum CMV loads were frequently more than 10-fold lower than plasma CMV loads.
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