Development and Evaluation of a Real-Time TaqMan-PCR for the Detection of Human Cytomegalovirus DNA in Bone Marrow Transplant Recipients

H Ghaffari; O Obeidi; M Dehghan; B Chahardouli; K Alimoghaddam; A Gharebaghian; AR Shamshiri; A Ghavamzadeh

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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

Author(s):

H Ghaffari^1,*,

O Obeidi²,

M Dehghan¹,

B Chahardouli¹,

K Alimoghaddam¹,

A Gharebaghian²,

AR Shamshiri¹,

A Ghavamzadeh¹

1Assistant Professor, Hematology, Oncology and Bone Marrow Transplantation Research Center; Tehran University of Medical Sciences, Tehran, Iran

2Research Assistant, Iran Blood Transfusion Organization Research Center, Tehran, Iran

Shiraz E-Medical Journal:Vol. 7, issue 3; e93680

Published online:Jul 01, 2006

Article type:Research Article

Received:May 14, 2019

Accepted:Jul 01, 2006

How to Cite:H GhaffariO ObeidiM DehghanB ChahardouliK AlimoghaddamA GharebaghianAR ShamshiriA Ghavamzadehet al.Development and Evaluation of a Real-Time TaqMan-PCR for the Detection of Human Cytomegalovirus DNA in Bone Marrow Transplant Recipients.Shiraz E-Med J.7(3):e93680.

Abstract

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.

Keywords

Cytomegalovirus

TaqMan PCR

pp65 Antigenemia

Bone Marrow Transplantation

References

1.
The references are available in the PDF file.

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