Detection of herpes simplex virus type-specific antibodies by an enzyme-linked immunosorbent assay based on glycoprotein G

Chelonid fibropapillomatosis-associated herpesvirus (CFPHV) is an alphaherpesvirus believed to cause marine turtle fibropapillomatosis (FP). A serodiagnostic assay was developed for monitoring sea turtle populations for CFPHV exposure. CFPHV glycoprotein H (gH) expressed in recombinant baculovirus was used in an enzyme-linked immunosorbent assay (ELISA) to detect virus-specific 7S turtle antibodies. Using captivereared green turtles (Chelonia mydas) with no history of virus exposure as "known negatives" and others with experimentally induced FP as "known positives," the assay had 100% specificity but low sensitivity, as seroconversion was detected in only half of the turtles bearing experimentally induced tumors. Antibodies were detected only in samples collected after cutaneous fibropapillomas appeared, consistent with observations that tumors are significant sites of virion production and antigen expression and the possibility that prolonged/ repeated virus shedding may be required for adequate stimulation of 7S antibody responses to gH. Natural routes of infection, however, may produce higher seroconversion rates. High gH antibody seroprevalences (ϳ80%) were found among wild green turtles in three Florida localities with different FP prevalences, including one site with no history of FP. In addition, all eight loggerhead turtles (Caretta caretta) tested were seropositive despite FP being uncommon in this species. The possibility that CFPHV infection may be common relative to disease suggests roles for environmental and host factors as modulators of disease expression. Alternatively, the possibility of other antigenically similar herpesviruses present in wild populations cannot be excluded, although antibody cross-reactivity with the lung/eye/trachea disease-associated herpesvirus was ruled out in this study.

show abstract

“…Nevertheless, serological assays that use well-characterized recom-VOL. 15,2008 ELISA FOR TURTLE ANTIBODIES TO CHELONID HERPESVIRUS gH 849…”

Section: Discussioncontrasting

confidence: 99%

“…In addition, glycoproteins may be relatively virus specific. For example, glycoprotein G has been found to distinguish antibody reactivity to the closely related human herpesviruses 1 and 2 (HSV-1 and HSV-2) (15,34).…”

mentioning

confidence: 99%

Use of Baculovirus-Expressed Glycoprotein H in an Enzyme-Linked Immunosorbent Assay Developed To Assess Exposure to Chelonid Fibropapillomatosis-Associated Herpesvirus and Its Relationship to the Prevalence of Fibropapillomatosis in Sea Turtles

Herbst

Lemaire

Ene

et al. 2008

Clin Vaccine Immunol

View full text Add to dashboard Cite

show abstract

“…Median follow-up time for Kalon nonseroconverters was 86 days (range, 58 to 139), while median follow-up time for Kalon seroconverters was over 3 months (median, 105 days; range, 47 to 193). Sensitivity for all three tests was lower in nonprimary than in primary patients (Table 1) in contrast to findings with other gG-2 assays (3,8), possibly due to the fact that follow-up time was shorter for nonprimary subjects (median, 81 days) than for primary patients (median, 104 days).…”

Section: The Centers For Disease Control and Prevention's Sexually Trmentioning

confidence: 99%

Performance of the Focus and Kalon Enzyme-Linked Immunosorbent Assays for Antibodies to Herpes Simplex Virus Type 2 Glycoprotein G in Culture-Documented Cases of Genital Herpes

Morrow

Friedrich

Krantz³

2003

J Clin Microbiol

View full text Add to dashboard Cite

Glycoprotein G-based herpes simplex virus type 2 (HSV-2) enzyme-linked immunosorbent assays from Focus and Kalon were performed with specimens from 118 patients with culture-documented genital herpes episodes, and their results were compared. Sensitivity was 52% by Kalon and 86% by Focus for first HSV-2 episodes and 100% (for each of the two tests) for recurrent HSV-2. Median times to seroconversion were 120 days by the Kalon assay, 21 days by the Focus assay, and 68 days by Western blotting assay. Values for specificity were 100% (Kalon) and 93% (Focus).

show abstract

“…The immunoaffinity-purified gG-2 was applied in an immunodot assay, and the lectin chromatography-purified gG-2 was used in an indirect enzyme-linked immunosorbent assay (ELISA). In addition, a capture ELISA (27) and a monoclonal blocking assay (23,50) were developed. Recombinant truncated gG produced in the baculovirus expression system was also used in an immunodot assay (46).…”

mentioning

confidence: 99%

Use of a Fragment of Glycoprotein G-2 Produced in the Baculovirus Expression System for Detecting Herpes Simplex Virus Type 2-Specific Antibodies

Ikoma¹,

Liljeqvist

Groen

et al. 2002

J Clin Microbiol

View full text Add to dashboard Cite

Fragments of glycoprotein G (gG-2 281-594His ), comprising residues 281 to 594 of herpes simplex virus type 2 (HSV-2), glycoprotein G of HSV-1 (gG-1 t26-189His ), and glycoprotein D of HSV-1 (gD-1 1-313 ), were expressed in the baculovirus expression system to develop an assay for the detection of HSV-1 and HSV-2 type-specific antibodies. The expression of the gG-1 t26-189His and gG-2 281-594His fragments was analyzed by Western blotting using monoclonal antibodies LP10 and AP1, respectively. The molecular masses of the major products of gG-1 t26-189His and the fragment of gG-2 281-594His were 36 to 39 kDa and 64 to 72 kDa, respectively. Human sera positive for HSV-1 reacted with gG-1 t26-189His , sera positive for HSV-2 reacted with the gG-2 281-594His fragment, and sera positive for both types reacted with gG-1 t26-189His and gG-2 281-594His in Western blotting. The human sera recognized polypeptides of gG-2 281-594His with molecular masses of 57 to 67 and 120 to 150 kDa and additional faint bands of 21, 29, and 45 kDa. The recombinant gG-1 t26-189His and the recombinant gG-2 281-594His fragment were used as type-specific antigens for the detection of HSV-1-and HSV-2-specific antibody responses in human sera, respectively. As type-common antigens, an extract of HSV-1-infected Vero cells and recombinant gD-1 1-313 were used. An enzyme-linked immunosorbent assay to detect type-specific antibodies was developed, and the sensitivity and specificity were evaluated by comparison with commercial tests by using sera obtained from different sources. The sensitivity and specificity were 91.5 and 95.5%, respectively, compared to the Gull assay. The gG-2 281-594His fragment can be obtained in relatively large quantities at low cost.Herpes simplex virus type 1 (HSV-1) and HSV-2 are well known as similar subtypes of HSV (38). HSV-2 is the main cause of recurrent genital infections (8). Most primary infections are asymptomatic, and people silently shed virus (14,30,51,57,58). After primary infection, the virus establishes latent infections in the local ganglia and is reactivated frequently, and antibody titers against HSV become detectable in serum samples. Many of the HSV-2 reactivations are asymptomatic and clinically not recognized (15,30,39,40,57). Most HSV infections are transmitted in the absence of lesions (17,57,58). Severe, generalized infections are seen particularly in neonates and immunocompromised (human immunodeficiency virus-infected) patients. Epidemiological studies indicate that in developed countries there is an ongoing HSV-2 epidemic with a significant rise in HSV-2 prevalence. Risk factors for acquiring HSV-2 infections are related to sexual life style, gender, race, and socioeconomic status. HSV type-specific antibody testing may be considered in a number of at-risk situations. Thus, identification of HSV-2-infected individuals may be important to reduce the risk of transmission in these groups.Laboratory diagnosis of HSV infections is based on direct detection of the virus and on serology (2, 16). Viru...

show abstract

Detection of herpes simplex virus type-specific antibodies by an enzyme-linked immunosorbent assay based on glycoprotein G

Cited by 25 publications

References 29 publications

Use of Baculovirus-Expressed Glycoprotein H in an Enzyme-Linked Immunosorbent Assay Developed To Assess Exposure to Chelonid Fibropapillomatosis-Associated Herpesvirus and Its Relationship to the Prevalence of Fibropapillomatosis in Sea Turtles

Use of Baculovirus-Expressed Glycoprotein H in an Enzyme-Linked Immunosorbent Assay Developed To Assess Exposure to Chelonid Fibropapillomatosis-Associated Herpesvirus and Its Relationship to the Prevalence of Fibropapillomatosis in Sea Turtles

Performance of the Focus and Kalon Enzyme-Linked Immunosorbent Assays for Antibodies to Herpes Simplex Virus Type 2 Glycoprotein G in Culture-Documented Cases of Genital Herpes

Use of a Fragment of Glycoprotein G-2 Produced in the Baculovirus Expression System for Detecting Herpes Simplex Virus Type 2-Specific Antibodies

Contact Info

Product

Resources

About