Tags

Type your tag names separated by a space and hit enter

Quantitation of cytomegalovirus DNA by real-time polymerase chain reaction in peripheral blood specimens of patients with solid organ transplants: comparison with end-point PCR and pp65 antigen test.
J Med Virol. 2006 Jul; 78(7):915-22.JM

Abstract

The polymerase chain reaction (PCR) for cytomegalovirus (CMV) DNA quantitation provides sensitive and specific data for detecting CMV as well as monitoring the infection and determining the appropriate antiviral strategy. A recently introduced real-time PCR assay for CMV DNA quantitation was applied on 158 peripheral blood leukocytes (PBLs) from 32 liver-transplanted patients with CMV asymptomatic infection and correlated with a commercial quantitative end-point PCR (COBAS AMPLICOR CMV Monitor) and CMV pp65 antigenemia. A good correlation was found between real-time PCR and pp65 antigen test (r2 = 0.691) and the two PCR assays (r2 = 0.761). Real-time PCR data were higher in pre-emptive treated patients (>20 pp65 + positive cells, median CMV DNA value: 3.8 log(10) copies/500,000 PBLs) than in not-treated ones (2.9 logs). According to pp65 levels of 0, 1-10, 11-20, 21-50, 51-100, and >100 positive cells/200,000 PBLs, median CMV DNA by real-time PCR was 2.6, 3.0, 3.6, 4.0, 4.2, and 4.8 logs, respectively, (CMV DNA levels by COBAS AMPLICOR: 2.8, 2.9, 3.8, 3.7, 3.9, and 4.0 logs). For samples with >20 pp65 + cells, real-time PCR gave significantly higher values than in groups with <20 pp65 + cells, whereas the COBAS AMPLICOR results showed a slower progression rate. Dilutions of CMV AD169 strain were used to probe real-time PCR reproducibility (between and intra-assay variability <2%) and sensitivity (100% detection rate at 10 copies/reaction, 28.5% with end-point PCR). In conclusion, real-time PCR significantly improves the study of CMV DNA dynamics due to a more reliable quantitation of CMV DNA for moderate and high DNA level compared to end-point PCR with better sensitivity and specificity. Real-time PCR provides more precise information for evaluating infection progress and assessing antiviral response, simplifying and accelerating the process of producing a reliable quantitation of CMV DNA for clinical purposes.

Authors+Show Affiliations

Microbiology Laboratory, Molinette Hospital, Turin, Italy.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Evaluation Study
Journal Article

Language

eng

PubMed ID

16721848

Citation

Allice, Tiziano, et al. "Quantitation of Cytomegalovirus DNA By Real-time Polymerase Chain Reaction in Peripheral Blood Specimens of Patients With Solid Organ Transplants: Comparison With End-point PCR and Pp65 Antigen Test." Journal of Medical Virology, vol. 78, no. 7, 2006, pp. 915-22.
Allice T, Enrietto M, Pittaluga F, et al. Quantitation of cytomegalovirus DNA by real-time polymerase chain reaction in peripheral blood specimens of patients with solid organ transplants: comparison with end-point PCR and pp65 antigen test. J Med Virol. 2006;78(7):915-22.
Allice, T., Enrietto, M., Pittaluga, F., Varetto, S., Franchello, A., Marchiaro, G., & Ghisetti, V. (2006). Quantitation of cytomegalovirus DNA by real-time polymerase chain reaction in peripheral blood specimens of patients with solid organ transplants: comparison with end-point PCR and pp65 antigen test. Journal of Medical Virology, 78(7), 915-22.
Allice T, et al. Quantitation of Cytomegalovirus DNA By Real-time Polymerase Chain Reaction in Peripheral Blood Specimens of Patients With Solid Organ Transplants: Comparison With End-point PCR and Pp65 Antigen Test. J Med Virol. 2006;78(7):915-22. PubMed PMID: 16721848.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Quantitation of cytomegalovirus DNA by real-time polymerase chain reaction in peripheral blood specimens of patients with solid organ transplants: comparison with end-point PCR and pp65 antigen test. AU - Allice,Tiziano, AU - Enrietto,Marco, AU - Pittaluga,Fabrizia, AU - Varetto,Silvia, AU - Franchello,Alessandro, AU - Marchiaro,Giovanna, AU - Ghisetti,Valeria, PY - 2006/5/25/pubmed PY - 2006/8/12/medline PY - 2006/5/25/entrez SP - 915 EP - 22 JF - Journal of medical virology JO - J Med Virol VL - 78 IS - 7 N2 - The polymerase chain reaction (PCR) for cytomegalovirus (CMV) DNA quantitation provides sensitive and specific data for detecting CMV as well as monitoring the infection and determining the appropriate antiviral strategy. A recently introduced real-time PCR assay for CMV DNA quantitation was applied on 158 peripheral blood leukocytes (PBLs) from 32 liver-transplanted patients with CMV asymptomatic infection and correlated with a commercial quantitative end-point PCR (COBAS AMPLICOR CMV Monitor) and CMV pp65 antigenemia. A good correlation was found between real-time PCR and pp65 antigen test (r2 = 0.691) and the two PCR assays (r2 = 0.761). Real-time PCR data were higher in pre-emptive treated patients (>20 pp65 + positive cells, median CMV DNA value: 3.8 log(10) copies/500,000 PBLs) than in not-treated ones (2.9 logs). According to pp65 levels of 0, 1-10, 11-20, 21-50, 51-100, and >100 positive cells/200,000 PBLs, median CMV DNA by real-time PCR was 2.6, 3.0, 3.6, 4.0, 4.2, and 4.8 logs, respectively, (CMV DNA levels by COBAS AMPLICOR: 2.8, 2.9, 3.8, 3.7, 3.9, and 4.0 logs). For samples with >20 pp65 + cells, real-time PCR gave significantly higher values than in groups with <20 pp65 + cells, whereas the COBAS AMPLICOR results showed a slower progression rate. Dilutions of CMV AD169 strain were used to probe real-time PCR reproducibility (between and intra-assay variability <2%) and sensitivity (100% detection rate at 10 copies/reaction, 28.5% with end-point PCR). In conclusion, real-time PCR significantly improves the study of CMV DNA dynamics due to a more reliable quantitation of CMV DNA for moderate and high DNA level compared to end-point PCR with better sensitivity and specificity. Real-time PCR provides more precise information for evaluating infection progress and assessing antiviral response, simplifying and accelerating the process of producing a reliable quantitation of CMV DNA for clinical purposes. SN - 0146-6615 UR - https://www.unboundmedicine.com/medline/citation/16721848/Quantitation_of_cytomegalovirus_DNA_by_real_time_polymerase_chain_reaction_in_peripheral_blood_specimens_of_patients_with_solid_organ_transplants:_comparison_with_end_point_PCR_and_pp65_antigen_test_ L2 - https://doi.org/10.1002/jmv.20641 DB - PRIME DP - Unbound Medicine ER -