Diagnostic value of measuring Epstein-Barr virus (EBV) DNA load and carcinoma-specific viral mRNA in relation to anti-EBV immunoglobulin A (IgA) and IgG antibody levels in blood of nasopharyngeal carcinoma patients from Indonesia

Servi J C Stevens; Sandra A W M Verkuijlen; Bambang Hariwiyanto; A Harijadi; Jajah Fachiroh; Dewi K Paramita; I Bing Tan; Sophia M Haryana; Jaap M Middeldorp

doi:https://doi.org/10.1128/JCM.43.7.3066-3073.2005

Diagnostic value of measuring Epstein-Barr virus (EBV) DNA load and carcinoma-specific viral mRNA in relation to anti-EBV immunoglobulin A (IgA) and IgG antibody levels in blood of nasopharyngeal carcinoma patients from Indonesia

Servi J C Stevens, Sandra A W M Verkuijlen, Bambang Hariwiyanto, A Harijadi, Jajah Fachiroh, Dewi K Paramita, I Bing Tan, Sophia M Haryana, Jaap M Middeldorp

Research output: Contribution to journal › Article › Academic › peer-review

75 Citations (Scopus)

Abstract

Nasopharyngeal carcinoma (NPC) is a prevalent malignancy in Southeast Asia and is strongly associated with Epstein-Barr virus (EBV). We investigated the primary diagnostic value of circulating EBV DNA and anti-EBV immunoglobulin G (IgG) and IgA levels in Indonesian NPC patients (n = 149). By a 213-bp Epstein-Barr virus nuclear antigen 1 (EBNA1)-based real-time LightCycler PCR, 72.5% of patients were positive for EBV DNA in whole blood, with 29.5% having levels above a previously determined clinical cutoff value (COV) of 2,000 EBV DNA copies/ml, the upper level in healthy carriers. In a 99-bp LightCycler PCR, 85.9% of patients were positive and 60.4% had levels above the COV. This assay quantified a significantly higher EBV load than the 213-bp PCR assay (P < 0.0001), suggesting that circulating EBV DNA is fragmented. Using data from 11 different studies, we showed a significant inverse correlation between PCR amplicon size and the percentage of patients positive for circulating EBV DNA (Spearman's rho = -0.91; P < 0.0001). EBV DNA loads were unrelated to anti-EBV IgG or IgA levels, as measured by VCA-p18 and EBNA1-specific synthetic peptide-based enzyme-linked immunosorbent assays. The presence of circulating tumor cells was assessed by amplification of BamHI-A rightward frame 1 (BARF1) mRNA, a viral oncogene abundantly expressed in EBV-carrying carcinomas but virtually absent from EBV-associated lymphomas. Despite high EBV DNA loads and the presence of EBNA1 and human U1A small nuclear ribonucleoprotein mRNA, BARF1 mRNA was never detected in blood. We conclude that amplicon size significantly influences EBV DNA load measurement in NPC patients. The circulating EBV DNA load is independent of serological parameters and does not reflect intact tumor cells. The primary diagnostic value of the EBV DNA load for the detection of NPC is limited.

Original language	English
Pages (from-to)	3066-3073
Number of pages	8
Journal	Journal of clinical microbiology
Volume	43
Issue number	7
DOIs	https://doi.org/10.1128/JCM.43.7.3066-3073.2005
Publication status	Published - Jul 2005

Keywords

Antibodies, Viral
Carcinoma
DNA, Viral
Evaluation Studies
Herpesvirus 4, Human
Humans
Immunoglobulin A
Immunoglobulin G
Indonesia
Journal Article
Nasopharyngeal Neoplasms
RNA, Messenger
Research Support, Non-U.S. Gov't
Viral Load

Access to Document

https://doi.org/10.1128/JCM.43.7.3066-3073.2005

Cite this

Stevens, S. J. C., Verkuijlen, S. A. W. M., Hariwiyanto, B., Harijadi, A., Fachiroh, J., Paramita, D. K., Tan, I. B., Haryana, S. M., & Middeldorp, J. M. (2005). Diagnostic value of measuring Epstein-Barr virus (EBV) DNA load and carcinoma-specific viral mRNA in relation to anti-EBV immunoglobulin A (IgA) and IgG antibody levels in blood of nasopharyngeal carcinoma patients from Indonesia. Journal of clinical microbiology, 43(7), 3066-3073. https://doi.org/10.1128/JCM.43.7.3066-3073.2005

Stevens, Servi J C ; Verkuijlen, Sandra A W M ; Hariwiyanto, Bambang et al. / Diagnostic value of measuring Epstein-Barr virus (EBV) DNA load and carcinoma-specific viral mRNA in relation to anti-EBV immunoglobulin A (IgA) and IgG antibody levels in blood of nasopharyngeal carcinoma patients from Indonesia. In: Journal of clinical microbiology. 2005 ; Vol. 43, No. 7. pp. 3066-3073.

@article{b824a092e6df43c9a30db3f85e6fbdc3,

title = "Diagnostic value of measuring Epstein-Barr virus (EBV) DNA load and carcinoma-specific viral mRNA in relation to anti-EBV immunoglobulin A (IgA) and IgG antibody levels in blood of nasopharyngeal carcinoma patients from Indonesia",

abstract = "Nasopharyngeal carcinoma (NPC) is a prevalent malignancy in Southeast Asia and is strongly associated with Epstein-Barr virus (EBV). We investigated the primary diagnostic value of circulating EBV DNA and anti-EBV immunoglobulin G (IgG) and IgA levels in Indonesian NPC patients (n = 149). By a 213-bp Epstein-Barr virus nuclear antigen 1 (EBNA1)-based real-time LightCycler PCR, 72.5% of patients were positive for EBV DNA in whole blood, with 29.5% having levels above a previously determined clinical cutoff value (COV) of 2,000 EBV DNA copies/ml, the upper level in healthy carriers. In a 99-bp LightCycler PCR, 85.9% of patients were positive and 60.4% had levels above the COV. This assay quantified a significantly higher EBV load than the 213-bp PCR assay (P < 0.0001), suggesting that circulating EBV DNA is fragmented. Using data from 11 different studies, we showed a significant inverse correlation between PCR amplicon size and the percentage of patients positive for circulating EBV DNA (Spearman's rho = -0.91; P < 0.0001). EBV DNA loads were unrelated to anti-EBV IgG or IgA levels, as measured by VCA-p18 and EBNA1-specific synthetic peptide-based enzyme-linked immunosorbent assays. The presence of circulating tumor cells was assessed by amplification of BamHI-A rightward frame 1 (BARF1) mRNA, a viral oncogene abundantly expressed in EBV-carrying carcinomas but virtually absent from EBV-associated lymphomas. Despite high EBV DNA loads and the presence of EBNA1 and human U1A small nuclear ribonucleoprotein mRNA, BARF1 mRNA was never detected in blood. We conclude that amplicon size significantly influences EBV DNA load measurement in NPC patients. The circulating EBV DNA load is independent of serological parameters and does not reflect intact tumor cells. The primary diagnostic value of the EBV DNA load for the detection of NPC is limited.",

keywords = "Antibodies, Viral, Carcinoma, DNA, Viral, Evaluation Studies, Herpesvirus 4, Human, Humans, Immunoglobulin A, Immunoglobulin G, Indonesia, Journal Article, Nasopharyngeal Neoplasms, RNA, Messenger, Research Support, Non-U.S. Gov't, Viral Load",

author = "Stevens, {Servi J C} and Verkuijlen, {Sandra A W M} and Bambang Hariwiyanto and A Harijadi and Jajah Fachiroh and Paramita, {Dewi K} and Tan, {I Bing} and Haryana, {Sophia M} and Middeldorp, {Jaap M}",

year = "2005",

month = jul,

doi = "https://doi.org/10.1128/JCM.43.7.3066-3073.2005",

language = "English",

volume = "43",

pages = "3066--3073",

journal = "Journal of clinical microbiology",

issn = "0095-1137",

publisher = "American Society for Microbiology",

number = "7",

}

Stevens, SJC, Verkuijlen, SAWM, Hariwiyanto, B, Harijadi, A, Fachiroh, J, Paramita, DK, Tan, IB, Haryana, SM & Middeldorp, JM 2005, 'Diagnostic value of measuring Epstein-Barr virus (EBV) DNA load and carcinoma-specific viral mRNA in relation to anti-EBV immunoglobulin A (IgA) and IgG antibody levels in blood of nasopharyngeal carcinoma patients from Indonesia', Journal of clinical microbiology, vol. 43, no. 7, pp. 3066-3073. https://doi.org/10.1128/JCM.43.7.3066-3073.2005

Diagnostic value of measuring Epstein-Barr virus (EBV) DNA load and carcinoma-specific viral mRNA in relation to anti-EBV immunoglobulin A (IgA) and IgG antibody levels in blood of nasopharyngeal carcinoma patients from Indonesia. / Stevens, Servi J C; Verkuijlen, Sandra A W M; Hariwiyanto, Bambang et al.
In: Journal of clinical microbiology, Vol. 43, No. 7, 07.2005, p. 3066-3073.

Research output: Contribution to journal › Article › Academic › peer-review

TY - JOUR

T1 - Diagnostic value of measuring Epstein-Barr virus (EBV) DNA load and carcinoma-specific viral mRNA in relation to anti-EBV immunoglobulin A (IgA) and IgG antibody levels in blood of nasopharyngeal carcinoma patients from Indonesia

AU - Stevens, Servi J C

AU - Verkuijlen, Sandra A W M

AU - Hariwiyanto, Bambang

AU - Harijadi, A

AU - Fachiroh, Jajah

AU - Paramita, Dewi K

AU - Tan, I Bing

AU - Haryana, Sophia M

AU - Middeldorp, Jaap M

PY - 2005/7

Y1 - 2005/7

N2 - Nasopharyngeal carcinoma (NPC) is a prevalent malignancy in Southeast Asia and is strongly associated with Epstein-Barr virus (EBV). We investigated the primary diagnostic value of circulating EBV DNA and anti-EBV immunoglobulin G (IgG) and IgA levels in Indonesian NPC patients (n = 149). By a 213-bp Epstein-Barr virus nuclear antigen 1 (EBNA1)-based real-time LightCycler PCR, 72.5% of patients were positive for EBV DNA in whole blood, with 29.5% having levels above a previously determined clinical cutoff value (COV) of 2,000 EBV DNA copies/ml, the upper level in healthy carriers. In a 99-bp LightCycler PCR, 85.9% of patients were positive and 60.4% had levels above the COV. This assay quantified a significantly higher EBV load than the 213-bp PCR assay (P < 0.0001), suggesting that circulating EBV DNA is fragmented. Using data from 11 different studies, we showed a significant inverse correlation between PCR amplicon size and the percentage of patients positive for circulating EBV DNA (Spearman's rho = -0.91; P < 0.0001). EBV DNA loads were unrelated to anti-EBV IgG or IgA levels, as measured by VCA-p18 and EBNA1-specific synthetic peptide-based enzyme-linked immunosorbent assays. The presence of circulating tumor cells was assessed by amplification of BamHI-A rightward frame 1 (BARF1) mRNA, a viral oncogene abundantly expressed in EBV-carrying carcinomas but virtually absent from EBV-associated lymphomas. Despite high EBV DNA loads and the presence of EBNA1 and human U1A small nuclear ribonucleoprotein mRNA, BARF1 mRNA was never detected in blood. We conclude that amplicon size significantly influences EBV DNA load measurement in NPC patients. The circulating EBV DNA load is independent of serological parameters and does not reflect intact tumor cells. The primary diagnostic value of the EBV DNA load for the detection of NPC is limited.

AB - Nasopharyngeal carcinoma (NPC) is a prevalent malignancy in Southeast Asia and is strongly associated with Epstein-Barr virus (EBV). We investigated the primary diagnostic value of circulating EBV DNA and anti-EBV immunoglobulin G (IgG) and IgA levels in Indonesian NPC patients (n = 149). By a 213-bp Epstein-Barr virus nuclear antigen 1 (EBNA1)-based real-time LightCycler PCR, 72.5% of patients were positive for EBV DNA in whole blood, with 29.5% having levels above a previously determined clinical cutoff value (COV) of 2,000 EBV DNA copies/ml, the upper level in healthy carriers. In a 99-bp LightCycler PCR, 85.9% of patients were positive and 60.4% had levels above the COV. This assay quantified a significantly higher EBV load than the 213-bp PCR assay (P < 0.0001), suggesting that circulating EBV DNA is fragmented. Using data from 11 different studies, we showed a significant inverse correlation between PCR amplicon size and the percentage of patients positive for circulating EBV DNA (Spearman's rho = -0.91; P < 0.0001). EBV DNA loads were unrelated to anti-EBV IgG or IgA levels, as measured by VCA-p18 and EBNA1-specific synthetic peptide-based enzyme-linked immunosorbent assays. The presence of circulating tumor cells was assessed by amplification of BamHI-A rightward frame 1 (BARF1) mRNA, a viral oncogene abundantly expressed in EBV-carrying carcinomas but virtually absent from EBV-associated lymphomas. Despite high EBV DNA loads and the presence of EBNA1 and human U1A small nuclear ribonucleoprotein mRNA, BARF1 mRNA was never detected in blood. We conclude that amplicon size significantly influences EBV DNA load measurement in NPC patients. The circulating EBV DNA load is independent of serological parameters and does not reflect intact tumor cells. The primary diagnostic value of the EBV DNA load for the detection of NPC is limited.

KW - Antibodies, Viral

KW - Carcinoma

KW - DNA, Viral

KW - Evaluation Studies

KW - Herpesvirus 4, Human

KW - Humans

KW - Immunoglobulin A

KW - Immunoglobulin G

KW - Indonesia

KW - Journal Article

KW - Nasopharyngeal Neoplasms

KW - RNA, Messenger

KW - Research Support, Non-U.S. Gov't

KW - Viral Load

U2 - https://doi.org/10.1128/JCM.43.7.3066-3073.2005

DO - https://doi.org/10.1128/JCM.43.7.3066-3073.2005

M3 - Article

C2 - 16002393

SN - 0095-1137

VL - 43

SP - 3066

EP - 3073

JO - Journal of clinical microbiology

JF - Journal of clinical microbiology

IS - 7

ER -

Stevens SJC, Verkuijlen SAWM, Hariwiyanto B, Harijadi A, Fachiroh J, Paramita DK et al. Diagnostic value of measuring Epstein-Barr virus (EBV) DNA load and carcinoma-specific viral mRNA in relation to anti-EBV immunoglobulin A (IgA) and IgG antibody levels in blood of nasopharyngeal carcinoma patients from Indonesia. Journal of clinical microbiology. 2005 Jul;43(7):3066-3073. doi: https://doi.org/10.1128/JCM.43.7.3066-3073.2005