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Multicentric evaluation of a new commercial cytomegalovirus real-time PCR quantitation assay.
J Virol Methods. 2007 Dec; 146(1-2):147-54.JV

Abstract

Automated real-time PCR systems have become the most common method in the quantitation of viral load during cytomegalovirus (CMV) infection in immuno-compromised patients. In order to evaluate a new commercially available CMV real-time PCR assay (CMV R-gene, Argene, France), a pp65 antigenemia assay and four different "in-house" real-time PCR assays were compared to the CMV R-gene for the detection and the quantitation of CMV load in 506 specimens of whole blood from transplant patients in four French hospital laboratories. The CMV R-gene was more sensitive than the pp65 antigenemia: there were 18% antigenemia-negative versus CMV R-gene-positive samples. A significant correlation was found between DNA quantitation by CMV R-gene and the number of positive cells detected by the pp65 antigenemia test (Spearman's rank test, r=0.63, p<0.0001). A CMV DNA load equivalent to 50 pp65-positive cells/200000 polymorphonuclear leukocytes was 5.26log(10)copies/mL of whole blood. When the CMV R-gene kit was compared to the four other "in-house" real-time PCR assays, there were few discordant results (6.7% total for the four laboratories), all detected with a weak positive CMV DNA viral load. Spearman's coefficients showed a good (r=0.82 for laboratory 1, r=0.66 for laboratory 3) to excellent (r=0.99 for laboratory 2, r=0.94 for laboratory 4) correlation between CMV R-gene and the four real-time "in-house" PCR assays. However, the results of CMV DNA viral load generated by CMV R-gene test were constantly higher than those generated by three out of four "in-house" PCR assays. This mean variation in CMV DNA viral load measured by CMV R-gene and "in-house" PCRs was of 0.77log(10), 0.04log(10), 0.77log(10) and 0.97log(10), for laboratories 1, 2, 3 and 4, respectively. We concluded that there was variability between results of different real-time PCR assays for CMV DNA quantitation. This observation emphasized the need of a standardised commercial assay to allow an "inter-laboratory" comparison of results. Our study showed that CMV R-gene is an accurate, efficient, reliable and versatile tool for rapid diagnosis and monitoring of CMV disease in transplantation recipients.

Authors+Show Affiliations

Laboratory of Virology, University Hospital, Avenue Georges Clemenceau, 14033 Caen Cedex, France. gouarin-s@chu-caen.frNo affiliation info availableNo affiliation info availableNo affiliation info availableNo 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
Multicenter Study

Language

eng

PubMed ID

17673304

Citation

Gouarin, S, et al. "Multicentric Evaluation of a New Commercial Cytomegalovirus Real-time PCR Quantitation Assay." Journal of Virological Methods, vol. 146, no. 1-2, 2007, pp. 147-54.
Gouarin S, Vabret A, Scieux C, et al. Multicentric evaluation of a new commercial cytomegalovirus real-time PCR quantitation assay. J Virol Methods. 2007;146(1-2):147-54.
Gouarin, S., Vabret, A., Scieux, C., Agbalika, F., Cherot, J., Mengelle, C., Deback, C., Petitjean, J., Dina, J., & Freymuth, F. (2007). Multicentric evaluation of a new commercial cytomegalovirus real-time PCR quantitation assay. Journal of Virological Methods, 146(1-2), 147-54.
Gouarin S, et al. Multicentric Evaluation of a New Commercial Cytomegalovirus Real-time PCR Quantitation Assay. J Virol Methods. 2007;146(1-2):147-54. PubMed PMID: 17673304.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Multicentric evaluation of a new commercial cytomegalovirus real-time PCR quantitation assay. AU - Gouarin,S, AU - Vabret,A, AU - Scieux,C, AU - Agbalika,F, AU - Cherot,J, AU - Mengelle,C, AU - Deback,C, AU - Petitjean,J, AU - Dina,J, AU - Freymuth,F, Y1 - 2007/07/27/ PY - 2007/03/26/received PY - 2007/06/14/revised PY - 2007/06/20/accepted PY - 2007/8/4/pubmed PY - 2008/1/26/medline PY - 2007/8/4/entrez SP - 147 EP - 54 JF - Journal of virological methods JO - J Virol Methods VL - 146 IS - 1-2 N2 - Automated real-time PCR systems have become the most common method in the quantitation of viral load during cytomegalovirus (CMV) infection in immuno-compromised patients. In order to evaluate a new commercially available CMV real-time PCR assay (CMV R-gene, Argene, France), a pp65 antigenemia assay and four different "in-house" real-time PCR assays were compared to the CMV R-gene for the detection and the quantitation of CMV load in 506 specimens of whole blood from transplant patients in four French hospital laboratories. The CMV R-gene was more sensitive than the pp65 antigenemia: there were 18% antigenemia-negative versus CMV R-gene-positive samples. A significant correlation was found between DNA quantitation by CMV R-gene and the number of positive cells detected by the pp65 antigenemia test (Spearman's rank test, r=0.63, p<0.0001). A CMV DNA load equivalent to 50 pp65-positive cells/200000 polymorphonuclear leukocytes was 5.26log(10)copies/mL of whole blood. When the CMV R-gene kit was compared to the four other "in-house" real-time PCR assays, there were few discordant results (6.7% total for the four laboratories), all detected with a weak positive CMV DNA viral load. Spearman's coefficients showed a good (r=0.82 for laboratory 1, r=0.66 for laboratory 3) to excellent (r=0.99 for laboratory 2, r=0.94 for laboratory 4) correlation between CMV R-gene and the four real-time "in-house" PCR assays. However, the results of CMV DNA viral load generated by CMV R-gene test were constantly higher than those generated by three out of four "in-house" PCR assays. This mean variation in CMV DNA viral load measured by CMV R-gene and "in-house" PCRs was of 0.77log(10), 0.04log(10), 0.77log(10) and 0.97log(10), for laboratories 1, 2, 3 and 4, respectively. We concluded that there was variability between results of different real-time PCR assays for CMV DNA quantitation. This observation emphasized the need of a standardised commercial assay to allow an "inter-laboratory" comparison of results. Our study showed that CMV R-gene is an accurate, efficient, reliable and versatile tool for rapid diagnosis and monitoring of CMV disease in transplantation recipients. SN - 0166-0934 UR - https://www.unboundmedicine.com/medline/citation/17673304/Multicentric_evaluation_of_a_new_commercial_cytomegalovirus_real_time_PCR_quantitation_assay_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0166-0934(07)00242-X DB - PRIME DP - Unbound Medicine ER -