scholarly journals Comparison of a Multiplexed Herpes Simplex Virus Type-Specific Immunoglobulin G Serology Assay to Immunoblot, Western Blot, and Enzyme-Linked Immunosorbent Assays

2008 ◽  
Vol 16 (1) ◽  
pp. 55-60 ◽  
Author(s):  
Thomas B. Martins ◽  
Ryan J. Welch ◽  
Harry R. Hill ◽  
Christine M. Litwin
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Western Blot ◽  
Immunoglobulin G ◽  
Multiplex Assay ◽  
Specific Alternative ◽  
Specific Antigens ◽  
Immunosorbent Assays ◽  
Simplex Virus ◽  
Hsv 1

ABSTRACT The human herpes simplex virus (HSV) is highly pathogenic, with infections caused by two distinct antigenic types, HSV-1 and HSV-2. Differentiation of antibodies to these specific antigens can provide useful information for the diagnosis of subclinical or undiagnosed HSV-2 infections, as well as for reducing the risk of maternal transfer of HSV to the neonate. In this study, a multiplex assay capable of concurrent detection of HSV-1 and -2 immunoglobulin G (IgG) antibodies was compared to immunoblot, Western blot, and enzyme-linked immunosorbent assays. Agreement of the multiplex assay was 95% or greater (n = 332) for both HSV-1 and -2 compared to the three assays. Sensitivities for HSV-1 ranged from 94.9 to 97.9%, with specificities of 93 to 97%. For HSV-2, the sensitivity and specificity ranges were 92.6 to 98.9% and 98.3 to 98.7%, respectively. Our studies show that the multiplexed microsphere-based assay offers a sensitive and specific alternative method for the detection HSV-1 and -2 type-specific antibodies. Advantages of the multiplex assay include multiple results per assay, the inclusion of internal controls for each specimen, and higher throughput of results.

scholarly journals Efficiency of Reconstitution of Immunoglobulin G from Blood Specimens Dried on Filter Paper and Utility in Herpes Simplex Virus Type-Specific Serology Screening

2002 ◽  
Vol 9 (6) ◽  
pp. 1338-1342 ◽  
Author(s):  
Wayne R. Hogrefe ◽  
Carolyn Ernst ◽  
Xin Su
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Immunoglobulin G ◽  
Herpes Simplex Virus Type ◽  
Filter Paper ◽  
Serum Samples ◽  
Blood Samples ◽  
The Mean ◽  
Simplex Virus ◽  
Hsv 1

ABSTRACT The performance of studies using sera from remote locations is greatly facilitated if whole-blood samples dried on filter paper are shown to be compatible with the serologic assay being employed. Since dried blood samples do not require immediate refrigeration, occupy little space, and are easily transported, they may be used for evaluating the seroprevalence of herpes simplex virus type 1 (HSV-1) and HSV-2 in geographic locations where laboratory resources are limited. We evaluated the utility of dried blood samples for the detection of type-specific HSV antibodies. The efficiency of using immunoglobulin G (IgG) eluted from dried blood samples was found to be consistent with measurement of IgG concentrations in most corresponding serum samples. The ratio of the mean IgG concentration for all dried blood samples to the mean IgG concentration for the corresponding sera was 1:29. When the 1:29 ratio was applied to each of the 22 pairs of samples, there was a deviation of less than 15% between concentrations in the dried blood sample and in the corresponding serum sample in 19 of the pairs. No positive or negative bias was detected for the IgG eluted from dried blood. The presence of HSV-1 and HSV-2 antibodies was determined in the paired dried blood and serum samples, and no differences in the HSV serostatuses were detected for 43 of the 44 pairs. One pair's serostatus varied, with the serum sample being weakly positive for HSV-1 and the dried blood sample results being equivocal. The detection of HSV antibodies was generally consistent for dried blood samples stored frozen for over 1 year or at room temperature for 30 days, although decreased reactivities were found in a few samples.

scholarly journals Limits in Reliability of Glycoprotein G-Based Type-Specific Serologic Assays for Herpes Simplex Virus Types 1 and 2

1999 ◽  
Vol 37 (2) ◽  
pp. 376-379 ◽  
Author(s):  
D. Scott Schmid ◽  
Denise R. Brown ◽  
Rosane Nisenbaum ◽  
Rae Lyn Burke ◽  
D’Anna Alexander ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Western Blot ◽  
Validation Studies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Western Blot Assay ◽  
Cross Sectional ◽  
Glycoprotein G ◽  
Simplex Virus ◽  
Hsv 1

Type-specific serologic assays for herpes simplex virus (HSV) types 1 and 2 based on glycoprotein G-1 (gG-1) (HSV-1) and gG-2 (HSV-2) discriminate between antibodies against HSV-1 and HSV-2. We previously developed a Western blot assay using gG-1 and gG-2 expressed in baculovirus, performed extensive validation studies, and determined that it was both sensitive and specific for type-specific detection of HSV antibody. Here we report that, among a cohort of Thai military recruits, the serostatus of some individuals changed from positive to negative over time (6.6% among those ever positive for HSV-1, and 14.9% among those ever positive for HSV-2). We tested a subset of these specimens in three other gG-based assays: an enzyme-linked immunosorbent assay, an immunoblot strip assay, and a Western blot assay. Positive-to-negative shifts occurred in every assay; the frequency of the shifts ranged from 6.1% to 21.2% of the specimen sets tested. There was only limited agreement among the assays concerning which individuals lost reactivity. This inaccuracy, exhibited by all of the assay protocols, was not predicted by validation studies employing specimens from cross-sectional studies and was most pronounced in HSV-2 testing. This argues for the inclusion of serial blood specimens in serologic assay validation procedures.

scholarly journals In Vivo Immune Evasion Mediated by the Herpes Simplex Virus Type 1 Immunoglobulin G Fc Receptor

1998 ◽  
Vol 72 (7) ◽  
pp. 5351-5359 ◽  
Author(s):  
Thandavarayan Nagashunmugam ◽  
John Lubinski ◽  
Liyang Wang ◽  
Lester T. Goldstein ◽  
Benjamin S. Weeks ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Immunoglobulin G ◽  
Mutant Virus ◽  
Wild Type Virus ◽  
Type Virus ◽  
Wild Type ◽  
Simplex Virus ◽  
Hsv 1

ABSTRACT Herpes simplex virus (HSV) glycoproteins gE and gI form an immunoglobulin G (IgG) Fc receptor (FcγR) that binds the Fc domain of human anti-HSV IgG and inhibits Fc-mediated immune functions in vitro. gE or gI deletion mutant viruses are avirulent, probably because gE and gI are also involved in cell-to-cell spread. In an effort to modify FcγR activity without affecting other gE functions, we constructed a mutant virus, NS-gE339, that has four amino acids inserted into gE within the domain homologous to mammalian IgG FcγRs. NS-gE339 expresses gE and gI, is FcγR−, and does not participate in antibody bipolar bridging since it does not block activities mediated by the Fc domain of anti-HSV IgG. In vivo studies were performed with mice because the HSV-1 FcγR does not bind murine IgG; therefore, the absence of an FcγR should not affect virulence in mice. NS-gE339 causes disease at the skin inoculation site comparably to wild-type and rescued viruses, indicating that the FcγR− mutant virus is pathogenic in animals. Mice were passively immunized with human anti-HSV IgG and then infected with mutant or wild-type virus. We postulated that the HSV-1 FcγR should protect wild-type virus from antibody attack. Human anti-HSV IgG greatly reduced viral titers and disease severity in NS-gE339-infected animals while having little effect on wild-type or rescued virus. We conclude that the HSV-1 FcγR enables the virus to evade antibody attack in vivo, which likely explains why antibodies are relatively ineffective against HSV infection.

scholarly journals Immunization Strategies To Block the Herpes Simplex Virus Type 1 Immunoglobulin G Fc Receptor

2004 ◽  
Vol 78 (5) ◽  
pp. 2562-2571 ◽  
Author(s):  
Xiaoqing Lin ◽  
John M. Lubinski ◽  
Harvey M. Friedman
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Virus Type ◽  
Immune Evasion ◽  
Immunoglobulin G ◽  
Herpes Simplex Virus Type ◽  
Fc Receptor ◽  
Simplex Virus ◽  
Hsv 1

ABSTRACT Herpes simplex virus type 1 (HSV-1) glycoprotein gE functions as an immunoglobulin G (IgG) Fc receptor (FcγR) that promotes immune evasion. When an IgG antibody binds by the F(ab′)2 domain to an HSV antigen, the Fc domain of some of the same antibody molecules binds to the FcγR, which blocks Fc-mediated functions. gE is a type 1 membrane glycoprotein with a large ectodomain that is expressed on the virion envelope and infected-cell surface. Our goal was to determine if immunizing with gE protein fragments could produce antibodies that bind by the F(ab′)2 domain to gE and block the FcγR, as measured by competitively inhibiting nonimmune human IgG binding to the FcγR. Three gE peptides were constructed in baculovirus spanning almost the entire ectodomain and used to immunize mice and rabbits. Two fragments were highly effective at producing antibodies that bind by the F(ab′)2 domain and block the FcγR. The most potent of these two antibodies was far more effective at blocking the FcγR than antibodies that are only capable of binding by the Fc domains to the FcγR, including anti-gC, anti-gD, and nonimmune IgG. These results suggest that immunizing with gE fragments has potential for preventing immune evasion by blocking activities mediated by the HSV-1 FcγR.

Herpes Simplex Virus and Human Cancer. III. Search for Relationship of Herpes Simplex Antibodies and Cervical Dysplasia and Labial Neoplasia

1983 ◽  
Vol 69 (2) ◽  
pp. 83-87 ◽  
Author(s):  
Antonella Rotola ◽  
Giuseppe Gerna ◽  
Dario Di Luca ◽  
Anna Rosa Virgili ◽  
Roberto Manservigi ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Antibody Titer ◽  
Human Cancer ◽  
Cervical Dysplasia ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Antigens ◽  
Simplex Virus ◽  
Relationship Of ◽  
Hsv 1

We employed the enzyme-linked immunosorbent assay (ELISA), indirect hemagglutination (IHA), and complement fixation (CF) methods to measure antibody titer to herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) in patients affected by labial tumors or cervical dysplasias. No relationship of antibody titer to HSV-1 and labial tumors was detected by any of the three methods. Association between antibody titer to HSV-2 and cervical dysplasias was revealed by IHA (p < 0.05) and ELISA (p < 0.001); CF tests were negative. Moreover, we assayed for HSV-specific antigens in cell cultures derived from labial tumors and cervical dysplasias. In cultures from labial tumors, it was not possible to detect HSV-specific antigens. Of the 25 cultures derived from cervical dysplasias, HSV antigens were found in only 3 cultures.

scholarly journals Multiplex immunoassay for detection of immunoglobulin G to herpes simplex virus types 1, 2 and cytomegalovirus based on PHOSPHAN technology

2017 ◽  
Vol 62 (2) ◽  
pp. 87-90
Author(s):  
A. V. Nikitina ◽  
V. G. Pomelova ◽  
N. S. Osin ◽  
S. G. Mardanly
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Immunoglobulin G ◽  
Clinical Laboratory ◽  
Recombinant Antigen ◽  
Multiplex Immunoassay ◽  
Standard Serum ◽  
Simplex Virus ◽  
Torch Infections ◽  
Hsv 1

We have developed a multiplex immunoassay test (immunochip) based on PHOSPHAN technology for the detection of immunoglobulin G to herpes simplex virus (HSV) types 1, 2 and cytomegalovirus (CMV). The immunochip consists of HSV type specific gG1 (HSV-1) and gG2 (HSV-2) recombinant antigens, the lysate antigen for detection of total IgG to both HSV types (HSV 1/2), and CMV specific chimeric recombinant antigen containing the immunodominant sequences of pp150, gB, pp28 and pp52 proteins. The sensitivity and specificity of simultaneous IgGs detection with recombinant proteins were comparable to the commercial ELISA kits regardless of the kind of investigated serum specimens (patient sera, standard serum panels). The lysate HSV antigen was as sensitive but significantly less specific, so that it could not be recommended for use as a component of the multiplex test. These results can be used as a basis for creating commercial multiplex tests intended for high-productive screening of HSV, CMV and other TORCH-infections in a clinical laboratory.

Assembly of VP26 in herpes simplex virus-1 capsid implicated by 3-D structure of recombinant capsid

1995 ◽  
Vol 53 ◽  
pp. 840-841
Author(s):  
Z. Hong Zhou ◽  
Jing He ◽  
Joanita Jakana ◽  
J. D. Tatman ◽  
Frazer J. Rixon ◽  
...  
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
3D Structure ◽  
Structure Study ◽  
Herpes Simplex Virus 1 ◽  
Shell Proteins ◽  
Life Threatening ◽  
Infected Cells ◽  
Simplex Virus ◽  
Hsv 1

Herpes simplex virus-1 (HSV-1) is a ubiquitous virus which is implicated in diseases ranging from self-curing cold sores to life-threatening infections. The 2500 Å diameter herpes virion is composed of a glycoprotein spike containing, lipid envelope, enclosing a protein layer (the tegument) in which is embedded the capsid (which contains the dsDNA genome). The B-, and A- and C-capsids, representing different morphogenetic stages in HSV-1 infected cells, are composed of 7, and 5 structural proteins respectively. The three capsid types are organized in similar T=16 icosahedral shells with 12 pentons, 150 hexons, and 320 connecting triplexes. Our previous 3D structure study at 26 Å revealed domain features of all these structural components and suggested probable locations for the outer shell proteins, VP5, VP26, VP19c and VP23. VP5 makes up most of both pentons and hexons. VP26 appeared to bind to the VP5 subunit in hexon but not to that in penton.

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