Shiraz E-Medical Journal
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Development and Evaluation of a Real-Time TaqMan-PCR for the Detection of Human Cytomegalovirus DNA in Bone Marrow Transplant Recipients
H Ghaffari1,*, O Obeidi2, M Dehghan1, B Chahardouli1, K Alimoghaddam1, A Gharebaghian2, AR Shamshiri1, A Ghavamzadeh1Abstract
Introduction: Cytomegalovirus (CMV) has been recognized as the most important viral pathogen in persons undergoing
bone marrow transplantation (BMT). In this study, we present the development of a TaqMan-based real-time PCR
assay to quantify human cytomegalovirus (CMV) DNA in peripheral blood leukocytes (PBLs) of bone marrow transplantation
patients.
Materials and Methods: A plasmid containing the target sequence from the pp65 region (UL83) of CMV was constructed
as a positive control template. Serial dilutions of 107 to 101 plasmids per assay were prepared. Peripheral blood
samples were collected from patients after transplantation. CMV DNA was quantified by RQ-PCR in parallel with the
pp65 antigenemia assay in PBL samples.
Results: The real-time PCR assay could detect CMV DNA in patient's samples with a wide linear range, from 10 to over
107 copies of CMV. Real-time PCR assay results correlated with those of the CMV pp65 antigenemia assay (P < 0.0001).
Discussion: The TaqMan assay may be a useful tool for rapid quantification of CMV infection and for monitoring of
CMV reactivation in bone marrow transplantation recipients. The results of both quantitative assays were significantly
correlated; however, the RQ-PCR assay was more sensitive than the pp65 antigenemia assay.
References
- 1.The references are available in the PDF file.
Copyright
© 2006, Author(s). This open-access article is available under the Creative Commons Attribution 4.0 (CC BY 4.0) International License (https://creativecommons.org/licenses/by/4.0/), which allows for unrestricted use, distribution, and reproduction in any medium, provided that the original work is properly cited.
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