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 ◽  
...  

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.

2022 ◽  
pp. 095646242110601
Author(s):  
Samer F Swedan ◽  
Alia’ Darabseh

Background Herpes infections are common infections among populations. Herein, a cross-sectional study was used to determine the seroprevalence of herpes simplex virus type 1 (HSV-1) and herpes simplex virus type 2 (HSV-2) IgG antibodies and their association with potential infection risk factors among Jordanians. Methods A total of 759 serum samples were collected (January to February 2020) and analyzed for HSV-1 and HSV-2 IgG antibodies by enzyme-linked immunosorbent assay. Estimates for population seropositivity were determined by weighting the age-specific seroprevalence by the size of the population in each age stratum. Results The population estimate for HSV-1 seroprevalence was 75.3%. After adjustment for possible confounders, regression analysis revealed higher seroprevalence with increase in age ( p < 0.005) and low household income ( p = 0.002). The population estimate for HSV-2 seroprevalence was 2.9%. No significant differences in HSV-2 seroprevalence were observed in association with age, gender, family size, educational level, and socioeconomic status, likely due to low seropositivity. Conclusions Jordanians have high HSV-1 and low HSV-2 seroprevalence. Periodical studies might be needed to evaluate changes in HSV-1 and HSV-2 seroprevalence over time. This study provides essential epidemiological data for Jordan and the Middle East and North Africa region.


2002 ◽  
Vol 9 (5) ◽  
pp. 1124-1125 ◽  
Author(s):  
Charles T. Leach ◽  
Rhoda L. Ashley ◽  
Jacques Baillargeon ◽  
Hal B. Jenson

ABSTRACT In 61 patients 1 to 14 years of age, the Gull/Meridian enzyme-linked immunosorbent assay (ELISA) had a sensitivity of 100% for herpes simplex virus type 1 (HSV-1) and specificities of 74% for HSV-1 and 48% for HSV-2. In 128 similarly aged patients, the HerpeSelect ELISA (Focus Technologies) showed sensitivities of 80% for HSV-1 and 88% for HSV-2, and specificities of 97% for HSV-1 and 100% for HSV-2.


1999 ◽  
Vol 6 (6) ◽  
pp. 826-831 ◽  
Author(s):  
Elham Rekabdar ◽  
Petra Tunbäck ◽  
Jan-Åke Liljeqvist ◽  
Tomas Bergström

ABSTRACT Glycoprotein G (gG) of herpes simplex virus type 1 (HSV-1) has been used as a prototype antigen for HSV-1 type-specific serodiagnosis, but data on the sequence variability of the gene coding for this protein in wild-type strains are lacking. In this study, direct DNA sequencing of the gG-1 genes from PCR products was performed with clinical HSV-1 isolates from 11 subjects as well as with strains Syn 17+, F, and KOS 321. The reference strains Syn 17+ and F showed a high degree of conservation, while KOS 321 carried 13 missense mutations and, in addition, 12 silent mutations. Three clinical isolates showed mutations leading to amino acid alterations: one had a mutation of K122 to N, which is a gG-1–to–gG-2 alteration; another contained all mutations which were observed in KOS 321 except two silent mutations; and the third isolate carried five missense mutations. Two clinical isolates as well as strain KOS 321 showed a mutation (F111→V) within the epitope of a gG-1-reactive monoclonal antibody (MAb). When all viruses were tested for reactivity with the anti-gG-1 MAb, the three strains with the F111→V mutation were found to be unreactive. Furthermore, gG-1 antibodies purified from sera from the two patients carrying strains mutated in this epitope were less reactive when they were tested by an HSV-1-infected-cell assay. Therefore, our finding that the sequence variability of the gG-1 gene also affects B-cell epitope regions of this protein in clinical isolates may have consequences for the use of this protein as a type-specific antigen for serodiagnosis.


1999 ◽  
Vol 37 (8) ◽  
pp. 2717-2718 ◽  
Author(s):  
Jan-Åke Liljeqvist ◽  
Bo Svennerholm ◽  
Tomas Bergström

The purpose of this study was to evaluate the performance of a herpes simplex virus (HSV) type 1-specific anti-glycoprotein C-1 monoclonal antibody (MAb) and a type 2-specific anti-glycoprotein G-2 MAb for typing of 2,400 clinical HSV-1 isolates and 2,400 clinical HSV-2 isolates, respectively, using an enzyme immunoassay. The anti-HSV-1 MAb showed sensitivity and specificity of 100%, and the anti-HSV-2 MAb showed a sensitivity of 99.46% and 100% specificity, indicating that these MAbs are suitable for typing of clinical HSV isolates.


2002 ◽  
Vol 83 (1) ◽  
pp. 157-165 ◽  
Author(s):  
Jan-Åke Liljeqvist ◽  
Edward Trybala ◽  
Johan Hoebeke ◽  
Bo Svennerholm ◽  
Tomas Bergström

Glycoprotein G-2 (gG-2) of herpes simplex virus type 2 (HSV-2) is cleaved to a secreted amino-terminal portion (sgG-2) and to a cell-associated carboxy-terminal portion which is further O-glycosylated to constitute the mature gG-2 (mgG-2). In contrast to mgG-2, which is known to elicit a type-specific antibody response in the human host, information on the immunogenic properties of sgG-2 is lacking. Here the sgG-2 protein was purified on a heparin column and used for production of monoclonal antibodies (mAbs). Four anti-sgG-2 mAbs were mapped using a Pepscan technique and identified linear epitopes which localized to the carboxy-terminal part of the protein. One additional anti-sgG-2 mAb, recognizing a non-linear epitope, was reactive to three discrete peptide stretches where the most carboxy-terminally located stretch was constituted by the amino acids 320RRAL323. Although sgG-2 is rapidly secreted into the cell-culture medium after infection, the anti-sgG-2 mAbs identified substantial amounts of sgG-2 in the cytoplasm of HSV-2-infected cells. All of the anti-sgG-2 mAbs were HSV-2 specific showing no cross-reactivity to HSV-1 antigen or to HSV-1-infected cells. Similarly, sera from 50 HSV-2 isolation positive patients were all reactive to sgG-2 in an enzyme immunoassay whilst no reactivity was seen in 25 sera from HSV-1 isolation positive patients or in 25 serum samples from HSV-negative patients suggesting that sgG-2 is a novel antigen potentially suitable for type-discriminating serodiagnosis.


2008 ◽  
Vol 16 (1) ◽  
pp. 55-60 ◽  
Author(s):  
Thomas B. Martins ◽  
Ryan J. Welch ◽  
Harry R. Hill ◽  
Christine M. Litwin

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.


2004 ◽  
Vol 85 (8) ◽  
pp. 2139-2147 ◽  
Author(s):  
Kristina Eriksson ◽  
Lars Bellner ◽  
Staffan Görander ◽  
Gun-Britt Löwhagen ◽  
Petra Tunbäck ◽  
...  

T-cell recognition of the secreted and membrane-bound portions of the herpes simplex virus type 2 (HSV-2) glycoprotein G (sgG-2 and mgG-2, respectively) was compared in symptomatic and asymptomatic HSV-2-infected individuals and in HSV-2-seronegative controls and the responses with HSV-1 glycoproteins C and E (gC-1 and gE-1) were compared. CD4+ T cells from HSV-2-infected individuals specifically recognized both sgG-2 and mgG-2, whereas HSV-1-infected and HSV-seronegative controls did not respond to these glycoproteins. The responses to gC-1 and gE-1, on the other hand, were not type specific, as blood mononuclear cells from both HSV-1- and HSV-2-infected individuals responded in vitro. There was an association between the status of the infection (symptomatic versus asymptomatic) and the CD4+ T-cell responsiveness. Symptomatic HSV-2-seropositive individuals responded with significantly lower Th1 cytokine production to sgG-2 and mgG-2 than did asymptomatic HSV-2-infected carriers, especially within the HSV-1-negative cohort. No differences in T-cell proliferation were observed between asymptomatic and symptomatic individuals. The results have implications for studies of HSV-2-specific CD4+ T-cell reactivity in general and for analysis of immunological differences between asymptomatic and symptomatic individuals in particular.


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