Measurement of Epstein-Barr virus DNA load using a novel quantification standard containing two EBV DNA targets and SYBR Green I dye

被引：28

作者：

Lay, Meav-Lang J. ^{[1
]}

Lucas, Robyn M. ^{[2
]}

Ratnamohan, Mala ^{[1
]}

Taylor, Janette ^{[1
]}

Ponsonby, Anne-Louise ^{[3
]}

Dwyer, Dominic E. ^{[1
]}

机构：

[1] Westmead Hosp, Dept Virol, Ctr Infect Dis & Microbiol Lab Serv, Inst Clin Pathol & Med Res, Westmead, NSW 2145, Australia

[2] Australian Natl Univ, Natl Ctr Epidemiol & Populat Hlth, Canberra, ACT 0200, Australia

[3] Royal Childrens Hosp, Murdoch Childrens Res Inst, Parkville, Vic 3052, Australia

来源：

VIROLOGY JOURNAL | 2010年 / 7卷

基金：

英国医学研究理事会;

关键词：

REAL-TIME PCR; POLYMERASE-CHAIN-REACTION; POSTTRANSPLANT LYMPHOPROLIFERATIVE DISORDER; STEM-CELL TRANSPLANTATION; QUANTITATIVE COMPETITIVE PCR; UNFRACTIONATED WHOLE-BLOOD; VIRAL LOAD; PERIPHERAL-BLOOD; NASOPHARYNGEAL CARCINOMA; HUMAN-HERPESVIRUS-6; DNA;

D O I：

10.1186/1743-422X-7-252

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

Background: Reactivation of Epstein-Barr virus (EBV) infection may cause serious, life-threatening complications in immunocompromised individuals. EBV DNA is often detected in EBV-associated disease states, with viral load believed to be a reflection of virus activity. Two separate real-time quantitative polymerase chain reaction (QPCR) assays using SYBR Green I dye and a single quantification standard containing two EBV genes, Epstein-Barr nuclear antigen-1 (EBNA-1) and BamHI fragment H rightward open reading frame-1 (BHRF-1), were developed to detect and measure absolute EBV DNA load in patients with various EBV-associated diseases. EBV DNA loads and viral capsid antigen (VCA) IgG antibody titres were also quantified on a population sample. Results: EBV DNA was measurable in ethylenediaminetetraacetic acid (EDTA) whole blood, peripheral blood mononuclear cells (PBMCs), plasma and cerebrospinal fluid (CSF) samples. EBV DNA loads were detectable from 8.0 x 10(2) to 1.3 x 10(8) copies/ml in post-transplant lymphoproliferative disease (n = 5), 1.5 x 10(3) to 2.0 x 10(5) copies/ml in infectious mononucleosis (n = 7), 7.5 x 10(4) to 1.1 x 10(5) copies/ml in EBV-associated haemophagocytic syndrome (n = 1), 2.0 x 10(2) to 5.6 x 10(3) copies/ml in HIV-infected patients (n = 12), and 2.0 x 10(2) to 9.1 x 10(4) copies/ml in the population sample (n = 218). EBNA-1 and BHRF-1 DNA were detected in 11.0% and 21.6% of the population sample respectively. There was a modest correlation between VCA IgG antibody titre and BHRF-1 DNA load (rho = 0.13, p = 0.05) but not EBNA-1 DNA load (rho = 0.11, p = 0.11). Conclusion: Two sensitive and specific real-time PCR assays using SYBR Green I dye and a single quantification standard containing two EBV DNA targets, were developed for the detection and measurement of EBV DNA load in a variety of clinical samples. These assays have application in the investigation of EBV-related illnesses in immunocompromised individuals.

引用

页数：11

共 76 条

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