Development of a Time-resolved Fluorescence Immunoassay for Epstein-Barr Virus Zta IgA Antibodies in Human Serum

Overview

Journal Viral Immunol

Specialties Allergy & Immunology
Microbiology

Date 2015 Feb 5

PMID 25651045

Authors

Juanjuan Chen

Tiancai Liu

Zhenhua Chen

Jingyuan Hou

Yingsong Wu

Ming Li

Affiliations

Soon will be listed here.

Abstract

The Epstein-Barr virus (EBV) transactivator protein (ZEBRA) is an immediate-early protein that plays an important role in the switch from latency to productive cycle in EBV virus. ZEBRA is an important marker of EBV reactivation. In order to diagnose EBV infection status correctly and timely, a novel immunoassay was developed based on an indirect time-resolved fluoroimmunoassay (TRFIA) for Zta IgA, which used recombinant Zta antigen as solid-phase antigen and Eu(3+)-labeled mouse antihuman IgA as corresponding probe. The precision, sensitivity, specificity test, and stability of the TRFIA kit were evaluated, and comparison with the traditional enzyme-linked immunosorbent assay (ELISA) was also investigated. The cutoff value for the TRFIA was 2.5. Intra- and interassay coefficients of variation for the TRFIA were 2.45-3.30% and 3.38-4.61% respectively. There was no cross-reactivity with the antibodies of cytomegalovirus (CMV) or herpes simplex virus (HSV) types 1 and 2, or other potential interferences. The established assay kit also behaved better in sensitivity and stability than the ELISA one. Additionally, the results in 382 serum samples using two analytical methods showed there was good agreement between the TRFIA and commercial ELISA kit. In the current study, the results demonstrated that the TRFIA that was developed for Zta IgA detection was more sensitive and reliable for the diagnosis of EBV infection and had potential value in automation and high-throughput screening.

References

Ng W, Yau T, Yung R, Sze W, Tsang A, Law A . Screening for family members of patients with nasopharyngeal carcinoma. Int J Cancer. 2004; 113(6):998-1001. DOI: 10.1002/ijc.20672. View

Yamauchi Y, Tachiband Y, Maeda A, Wakiguchi H, Usui M, Kurata T . Evaluation of antibodies to the Epstein-Barr virus immediate early gene product ZEBRA by a new enzyme-linked immunosorbent assay. Intervirology. 1999; 41(6):278-84. DOI: 10.1159/000024950. View

Hou J, Liu T, Lin G, Li Z, Zou L, Li M . Development of an immunomagnetic bead-based time-resolved fluorescence immunoassay for rapid determination of levels of carcinoembryonic antigen in human serum. Anal Chim Acta. 2012; 734:93-8. DOI: 10.1016/j.aca.2012.04.044. View

Henle W, Henle G, Lennette E . The Epstein-Barr virus. Sci Am. 1979; 241(1):48-59. DOI: 10.1038/scientificamerican0779-48. View

Bengali Z, Das S, Middleton M, Levine P . Seroepidemiology of Epstein--Barr virus-associated diseases--I. A pilot evaluation using a radiometric quantitative complement fixation test. Comp Immunol Microbiol Infect Dis. 1979; 2(2-3):213-20. DOI: 10.1016/0147-9571(79)90009-2. View

Dopatka H, Schuy W . Compact Epstein-Barr virus diagnosis based on a defined antigen mix and specific IgA. Res Virol. 1996; 147(1):53-66. DOI: 10.1016/0923-2516(96)80240-8. View

Countryman J, Miller G . Activation of expression of latent Epstein-Barr herpesvirus after gene transfer with a small cloned subfragment of heterogeneous viral DNA. Proc Natl Acad Sci U S A. 1985; 82(12):4085-9. PMC: 397939. DOI: 10.1073/pnas.82.12.4085. View

Yip T, Ngan R, Lau W, Poon Y, Joab I, Cochet C . A possible prognostic role of immunoglobulin-G antibody against recombinant Epstein-Barr virus BZLF-1 transactivator protein ZEBRA in patients with nasopharyngeal carcinoma. Cancer. 1994; 74(9):2414-24. DOI: 10.1002/1097-0142(19941101)74:9<2414::aid-cncr2820740905>3.0.co;2-8. View

Niedobitek G, Young L . Epstein-Barr virus persistence and virus-associated tumours. Lancet. 1994; 343(8893):333-5. DOI: 10.1016/s0140-6736(94)91167-3. View

10.

Drouet E, Brousset P, Fares F, Icart J, Verniol C, Meggetto F . High Epstein-Barr virus serum load and elevated titers of anti-ZEBRA antibodies in patients with EBV-harboring tumor cells of Hodgkin's disease. J Med Virol. 1999; 57(4):383-9. DOI: 10.1002/(sici)1096-9071(199904)57:4<383::aid-jmv10>3.0.co;2-3. View

11.

Fachiroh J, Paramita D, Hariwiyanto B, Harijadi A, Dahlia H, Indrasari S . Single-assay combination of Epstein-Barr Virus (EBV) EBNA1- and viral capsid antigen-p18-derived synthetic peptides for measuring anti-EBV immunoglobulin G (IgG) and IgA antibody levels in sera from nasopharyngeal carcinoma patients: options for.... J Clin Microbiol. 2006; 44(4):1459-67. PMC: 1448657. DOI: 10.1128/JCM.44.4.1459-1467.2006. View

12.

Tedeschi R, Dillner J, De Paoli P . Antibody responses to defined epitopes in the Epstein-Barr virus BZLF1-encoded transactivator protein among human immunodeficiency virus-infected patients. Clin Diagn Lab Immunol. 1996; 3(3):363-4. PMC: 170349. DOI: 10.1128/cdli.3.3.363-364.1996. View

13.

Lin G, Liu T, Zou L, Hou J, Wu Y . Development of a dual-label time-resolved fluoroimmunoassay for the detection of α-fetoprotein and hepatitis B virus surface antigen. Luminescence. 2012; 28(3):401-6. DOI: 10.1002/bio.2396. View

14.

Countryman J, Jenson H, Seibl R, Wolf H, Miller G . Polymorphic proteins encoded within BZLF1 of defective and standard Epstein-Barr viruses disrupt latency. J Virol. 1987; 61(12):3672-9. PMC: 255978. DOI: 10.1128/JVI.61.12.3672-3679.1987. View

15.

Gan Y, Chan S, Gan L . Epstein-Barr Viral Antigens Used in the Diagnosis of Nasopharyngeal Carcinoma. J Biomed Sci. 1996; 3(3):159-169. DOI: 10.1007/BF02253096. View

16.

Bethony J, Brooker S, Albonico M, Geiger S, Loukas A, Diemert D . Soil-transmitted helminth infections: ascariasis, trichuriasis, and hookworm. Lancet. 2006; 367(9521):1521-32. DOI: 10.1016/S0140-6736(06)68653-4. View

17.

Klein G . Epstein-Barr virus strategy in normal and neoplastic B cells. Cell. 1994; 77(6):791-3. DOI: 10.1016/0092-8674(94)90125-2. View

18.

Joab I, Nicolas J, Schwaab G, Clausse B, Perricaudet M, Zeng Y . Detection of anti-Epstein-Barr-virus transactivator (ZEBRA) antibodies in sera from patients with nasopharyngeal carcinoma. Int J Cancer. 1991; 48(5):647-9. DOI: 10.1002/ijc.2910480503. View

19.

Zhang X, Zhang Y, Nie Y, Wang S, Chen Y, Sun D . Serum Zta antibody of Epstein-Barr virus exerts potential function in the diagnosis of nasopharyngeal cancer. Tumour Biol. 2014; 35(7):6879-86. DOI: 10.1007/s13277-014-1940-y. View

20.

Fachiroh J, Prasetyanti P, Paramita D, Prasetyawati A, Anggrahini D, Haryana S . Dried-blood sampling for epstein-barr virus immunoglobulin G (IgG) and IgA serology in nasopharyngeal carcinoma screening. J Clin Microbiol. 2008; 46(4):1374-80. PMC: 2292939. DOI: 10.1128/JCM.01368-07. View