Detection of anti-hepatitis C virus antibodies by ELISA using synthetic peptides

Berasain, Carmen; Garcı́a-Granero, Marta; Riezu-Boj, José Ignacio; Civeira, M.P.; Prieto, Jesús; Borrás‐Cuesta, Francisco

doi:10.1016/s0168-8278(05)80013-3

Cited by 6 publications

(4 citation statements)

References 28 publications

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“…However, there is insufficient information regarding antibody responses to nonstructural proteins even though they are involved in the pathogenesis of the disease. The antigens used in most of the reported works were prepared from tissue cultures (Berasain et al 1993, Videa et al 2005. This is laborious, expensive, and subject to batch-to-batch quality variation which makes it unsuitable for routine large-scale production, especially in tropical undeveloped countries in which the disease is endemic.…”

Section: Discussionmentioning

confidence: 99%

“…However, detection of the antibody response to other nonstructural proteins with different clinical features and types of infections may provide information about the diagnosis, prognosis, or immunopathology of DF. Regarding this point, some recombinant proteins have been used successfully to evaluate antibody responses, such as the core (C), NS3, NS4, and NS5 proteins of hepatitis C, which is a member of the Flavivirus family (Berasain et al 1993). Wong et al (2003) used recombinant NS3 and NS5 proteins to develop a serological assay for West Nile virus diagnosis and discriminated between West Nile infections and DEN or St. Louis encephalitis virus infections.…”

Section: Discussionmentioning

confidence: 99%

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Analysis of Antibody Response in Human Dengue Patients from the Mexican Coast Using Recombinant Antigens

Lázaro-Olan

Mellado-Sánchez²,

García‐Cordero

et al. 2008

Vector-Borne and Zoonotic Diseases

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This study was undertaken to evaluate the feasibility of using recombinant dengue proteins to discriminate between acute dengue infections versus uninfected dengue samples. Dengue virus proteins E, NS1, NS3, and NS4B were cloned as fusion proteins and expressed in Escherichia coli. Recombinant products were tested in 100 serum samples obtained from acute dengue fever cases collected from 3 states of Mexico where dengue is endemic. Sera from 75 healthy individuals living in nonendemic areas for dengue were used as a control group. In sera from the dengue patients group, antibody responses to E protein were demonstrated in 91% of cases and NS1 protein was recognized to various extents (99%) within the first 7 days of infection. The antibody responses to NS3 and NS4B were frequently of low magnitude. Consistent negative antibody responses to all proteins were found in sera from the control group. These data suggest that the glutathione-S-transferase (GST)-dengue fusion proteins may be feasible antigens for a sensitive and specific serological assay.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Analysis of Antibody Response in Human Dengue Patients from the Mexican Coast Using Recombinant Antigens

Lázaro-Olan

Mellado-Sánchez²,

García‐Cordero

et al. 2008

Vector-Borne and Zoonotic Diseases

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“…Investigations into viral non-structural proteins could yield more accurate information about their potential role in immunogenicity and lead to the development of a diagnostic tool and further unravelling of the immunopathology of the disease in different types of infection and clinical manifestations. In this regard, several non-structural proteins have been successfully exploited to evaluate antibody responses to viral infections belonging to family Flaviviridae and led to the differentiation of different diseases stages and clinical manifestations [10,[28][29][30][31]. Since NS4B plays a significant role in eliciting humoral and cellular immunity in other viral infections, initial commercial diagnostic assays were based on this protein due to harboring of highly conserved epitopes of diagnostic significance [32][33][34].…”

Section: Discussionmentioning

confidence: 99%

Deciphering the Role of Bovine Viral Diarrhea Virus Non-Structural NS4B Protein in Viral Pathogenesis

Bashir

Kossarev

Martin

et al. 2020

Veterinary Sciences

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Bovine viral diarrhea virus (BVDV) is a (+) ssRNA virus that belongs to the family Flaviviridae. BVDV is a significant animal pathogen causing substantial economic losses to the cattle industry worldwide through respiratory and gastrointestinal infections and abortion or birth of persistently infected calves. While the immunogenic profile of some of the BVDV proteins (i.e., Erns, E2 and NS3) is well established during viral pathogenesis, very little information is available about most of BVDV’s non-structural proteins in this regard. In recent times, the NS4B protein has emerged as an interesting target of diagnostic, vaccination and therapeutic value in viral infections of other members of the family Flaviviridae due to its key scaffold-like contribution in the viral replication complex. Although, BVDV-NS4B has a membrane topology alongside its role in induction of autophagosomes in vitro. However, information on its immunogenicity during BVDV pathogenesis and vaccination is scarce. To characterize the immunogenic profile of the NS4B, five cows were vaccinated with the live attenuated BVDV vaccine Bovela® and blood samples were taken pre- and post-immunization for serum isolation. Virus neutralization assay (VNA) confirmed the presence of anti-BVDV antibodies in the sera of vaccinated cows. VNA also revealed pre-existing antibodies against BVDV in the pre-immunization sera of two cows. To identify BVDV-NS4B specific antibodies, the NS4B protein was expressed in mammalian cells by using the pCI-neo vector system. The sera from BVDV vaccinated cows were evaluated for the presence of BVDV-NS4B specific antibodies through western blot and indirect ELISA. Interestingly, t sera from cows with pre-existing immunity against BVDV were able to detect NS4B in western blot and ELISA, suggesting the presence of NS4B-specific antibodies. The obtained results provide the first indication of the immunogenic nature of BVDV-NS4B protein in sero-converted animals. These findings are consistent with the observation made for NS4B in other Flaviviridae members and confirm this protein as an interesting target with diagnostic, vaccination and therapeutic value.

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“…A number of diagnostically relevant antigenic epitopes have been found within the core region [Muraiso et al, 1990;Chiba et al, 1991;Hosein et al, 1991;Nasoff et al, 1991;Sallberg et al, 1992], E1/E2 [Lok et al, 1993;Selby et al, 1993;Ray et al, 1994;], NS3 [Chien et al, 1992;], NS4 Chien et al, 1992;] and NS5 proteins [ Mori, 1992;Riezu-Boj, 1992]. The HCV core, NS3 and NS4 proteins are the most diagnostically relevant antigens now used in various commercial immunoassays [ Muraiso et al, 1990;Hosein et al, 1991;Van Der Poel et al, 1991;Chien et al, 1992;Inoue et al, 1992;Mori et al, 1992;Riezu-Boj et al, 1992;Berasain et al, 1993;Bukh, 1995]. The NS4 recombinant protein c100-3 of genotype 1 was shown less immunoreactive or nonimmunoreactive with antibodies in the serum specimens from the patients infected with HCV genotypes 2 and 3 compared to the immunoreactivity with the serum specimens from patients infected with genotype 1 [McOmish et al, 1993].…”

Section: Introductionmentioning

confidence: 99%

Artificial NS4 mosaic antigen of hepatitis C virus

et al. 1999

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An artificial antigen composed of 17 small antigenic regions derived from the NS4-protein of hepatitis C virus (HCV) genotypes 1 through 5 was designed and constructed. Eleven antigenic regions were derived from the 5-1-1 region, and 6 others were derived from the C-terminus of the NS4-protein of different genotypes. The gene encoding for this artificial antigen was assembled from synthetic oligonucleotides by a new approach designated as restriction enzyme-assisted ligation (REAL). The full-length synthetic gene was expressed in Escherichia coli as a fusion protein with glutathione S-transferase. By the use of site-specific antibodies raised against synthetic peptides, it was shown that all regions for which sequence-specific antibodies were obtained were accessible to antibody binding. The diagnostic relevance of the NS4 artificial antigen was demonstrated by testing this antigen with 4 HCV seroconversion panels and a panel of previously tested and stored serum specimens. The artificial antigen was found to specifically detect anti-NS4 antibodies in a number of specimens that were previously found to be anti-NS4 negative. Furthermore, this antigen detected anti-NS4 activity earlier in 2 of 4 seroconversion panels than did the antigen used in a commercially available supplemental assay. Equally important is the observation that the artificial NS4 antigen demonstrated equivalent anti-NS4 immunoreactivity with serum specimens obtained from patients infected with different HCV genotypes, whereas the NS4 recombinant protein derived from genotype 1, used in the commercial supplemental test, was less immunoreactive with serum specimens containing HCV genotypes 2, 3, and 4. Collectively, these data support the significant diagnostic potential of the NS4 mosaic antigen. The strategy employed in this study may be applied to the design and construction of other artificial antigens with improved diagnostically pertinent properties. J. Med. Virol. 59:437-450 1999.

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Detection of anti-hepatitis C virus antibodies by ELISA using synthetic peptides

Cited by 6 publications

References 28 publications

Analysis of Antibody Response in Human Dengue Patients from the Mexican Coast Using Recombinant Antigens

Analysis of Antibody Response in Human Dengue Patients from the Mexican Coast Using Recombinant Antigens

Deciphering the Role of Bovine Viral Diarrhea Virus Non-Structural NS4B Protein in Viral Pathogenesis

Artificial NS4 mosaic antigen of hepatitis C virus

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