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Whole blood real-time quantitative PCR for cytomegalovirus infection follow-up in transplant recipients.
J Clin Virol. 2006 May; 36(1):72-5.JC

Abstract

BACKGROUND

Cytomegalovirus (CMV) remains a major opportunistic agent among transplant recipients. While detection of CMV pp65-lower matrix protein (pp65Ag) is still widely used for monitoring CMV infection, real-time PCR assays have been recently developed for routine quantitation of CMV DNA. However, correlations are lacking between results of pp65Ag and quantitative PCR assays and there is no consensus yet as to the more appropriate blood compartment (whole blood (WB), leukocytes, plasma) to be tested with PCR assays.

OBJECTIVES

The aims of the study were to determine, in a population of transplant recipients: (i) the correlation between pp65Ag and CMV quantitative real-time PCR in our setting and (ii) the utility of plasma CMV DNA quantitation in comparison to WB quantitation.

METHODS

In 170 blood samples (from 61 solid organ or bone marrow transplant recipients) with pp65Ag results, CMV quantitation was performed in WB and plasma using an in-house real-time quantitative PCR.

RESULTS

Real-time PCR and pp65Ag results in WB were correlated: thresholds of 10 and 50(+) cells/200,000 cells were equivalent to 3.3 log(10)copies/mL (2,000 copies/mL) and 3.8 log(10)copies/mL (6,300 copies/mL), respectively. When WB viral load was >or=3.6 log(10)copies/mL, the risk to have a negative plasma CMV DNA result was <or=2.5%.

CONCLUSIONS

For the routine exploration of a single compartment, whole blood would represent a suitable compromise: easy processing for a sensitive assay. The 3.6 log(10)copies/mL threshold, which could help in identifying active CMV infection, deserves further evaluation.

Authors+Show Affiliations

Département de Virologie et d'Immunologie Biologique and Laboratoire EA 2968, Centre Hospitalo-Universitaire de Bordeaux, 33076 Bordeaux, France.No 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
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16481215

Citation

Garrigue, Isabelle, et al. "Whole Blood Real-time Quantitative PCR for Cytomegalovirus Infection Follow-up in Transplant Recipients." Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology, vol. 36, no. 1, 2006, pp. 72-5.
Garrigue I, Boucher S, Couzi L, et al. Whole blood real-time quantitative PCR for cytomegalovirus infection follow-up in transplant recipients. J Clin Virol. 2006;36(1):72-5.
Garrigue, I., Boucher, S., Couzi, L., Caumont, A., Dromer, C., Neau-Cransac, M., Tabrizi, R., Schrive, M. H., Fleury, H., & Lafon, M. E. (2006). Whole blood real-time quantitative PCR for cytomegalovirus infection follow-up in transplant recipients. Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology, 36(1), 72-5.
Garrigue I, et al. Whole Blood Real-time Quantitative PCR for Cytomegalovirus Infection Follow-up in Transplant Recipients. J Clin Virol. 2006;36(1):72-5. PubMed PMID: 16481215.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Whole blood real-time quantitative PCR for cytomegalovirus infection follow-up in transplant recipients. AU - Garrigue,Isabelle, AU - Boucher,Sébastien, AU - Couzi,Lionel, AU - Caumont,Anne, AU - Dromer,Claire, AU - Neau-Cransac,Martine, AU - Tabrizi,Reza, AU - Schrive,Marie-Hélène, AU - Fleury,Hervé, AU - Lafon,Marie-Edith, Y1 - 2006/02/14/ PY - 2005/12/23/received PY - 2006/01/05/revised PY - 2006/01/06/accepted PY - 2006/2/17/pubmed PY - 2006/6/6/medline PY - 2006/2/17/entrez SP - 72 EP - 5 JF - Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology JO - J Clin Virol VL - 36 IS - 1 N2 - BACKGROUND: Cytomegalovirus (CMV) remains a major opportunistic agent among transplant recipients. While detection of CMV pp65-lower matrix protein (pp65Ag) is still widely used for monitoring CMV infection, real-time PCR assays have been recently developed for routine quantitation of CMV DNA. However, correlations are lacking between results of pp65Ag and quantitative PCR assays and there is no consensus yet as to the more appropriate blood compartment (whole blood (WB), leukocytes, plasma) to be tested with PCR assays. OBJECTIVES: The aims of the study were to determine, in a population of transplant recipients: (i) the correlation between pp65Ag and CMV quantitative real-time PCR in our setting and (ii) the utility of plasma CMV DNA quantitation in comparison to WB quantitation. METHODS: In 170 blood samples (from 61 solid organ or bone marrow transplant recipients) with pp65Ag results, CMV quantitation was performed in WB and plasma using an in-house real-time quantitative PCR. RESULTS: Real-time PCR and pp65Ag results in WB were correlated: thresholds of 10 and 50(+) cells/200,000 cells were equivalent to 3.3 log(10)copies/mL (2,000 copies/mL) and 3.8 log(10)copies/mL (6,300 copies/mL), respectively. When WB viral load was >or=3.6 log(10)copies/mL, the risk to have a negative plasma CMV DNA result was <or=2.5%. CONCLUSIONS: For the routine exploration of a single compartment, whole blood would represent a suitable compromise: easy processing for a sensitive assay. The 3.6 log(10)copies/mL threshold, which could help in identifying active CMV infection, deserves further evaluation. SN - 1386-6532 UR - https://www.unboundmedicine.com/medline/citation/16481215/Whole_blood_real_time_quantitative_PCR_for_cytomegalovirus_infection_follow_up_in_transplant_recipients_ DB - PRIME DP - Unbound Medicine ER -