SARS Antibody Test for Serosurveillance

Hsueh, Po-Ren; Kao, Chuan-Liang; Lee, Chun Nan; Chen, Li Kuan; Ho, Margaret S.; Sia, Charles; Fang, Xin; Lynn, Shugene; Chang, Tseng Yuan; Liu, Shi Kau; Walfield, Alan M.; Wang, Chang Yi

doi:10.3201/eid1009.040101

Cited by 31 publications

(28 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For SARS-CoV infection, it has been shown that nucleotides 952-1530 of the spike protein gene of SARS-CoV encoded a 193-amino acid fragment responsible for attaching to the receptor for SARS-CoV, angiotensin-converting enzyme 2 [9]. Furthermore, we, and also others, have shown that patients with SARS produced antibody response against the spike protein of SARS-CoV [3,10,11], and it has been demonstrated that the spike protein is the major target for passive immunization [12,13]. In studies that determine the relative importance of humoral and cell mediated immunity for protection against SARS-CoV infection, it was confirmed that neutralizing antibody, when administered by passive immunization, was crucial in conferring protection [14], whereas T-cell immunity was unable to lead to protection [15].…”

Section: Introductionmentioning

confidence: 51%

SARS coronavirus spike polypeptide DNA vaccine priming with recombinant spike polypeptide from Escherichia coli as booster induces high titer of neutralizing antibody against SARS coronavirus

Woo

Lau

Tsoi

et al. 2005

Vaccine

View full text Add to dashboard Cite

Different forms of SARS coronavirus (SARS-CoV) spike protein-based vaccines for generation of neutralizing antibody response against SARS-CoV were compared using a mouse model. High IgG levels were detected in mice immunized with intraperitoneal (i.p.) recombinant spike polypeptide generated by Escherichia coli (S-peptide), mice primed with intramuscular (i.m.) tPA-optimize800 DNA vaccine (tPA-S-DNA) and boosted with i.p. S-peptide, mice primed with i.m. CTLA4HingeSARS800 DNA vaccine (CTLA4-S-DNA) and boosted with i.p. S-peptide, mice primed with oral live-attenuated Salmonella typhimurium (Salmonella-S-DNA-control) and boosted with i.p. S-peptide, mice primed with oral live-attenuated S. typhimurium that contained tPA-optimize800 DNA vaccine (Salmonella-tPA-S-DNA) and boosted with i.p. S-peptide, and mice primed with oral live-attenuated S. typhimurium that contained CTLA4HingeSARS800 DNA vaccine (Salmonella-tPA-S-DNA) and boosted with i.p. S-peptide. No statistical significant difference was observed among the Th1/Th2 index among these six groups of mice with high IgG levels. Sera of all six mice immunized with i.p. S-peptide, i.m. DNA vaccine control and oral Salmonella-S-DNA-control showed no neutralizing antibody against SARS-CoV. Sera of the mice immunized with i.m. tPA-S-DNA, i.m. CTLA4-S-DNA, oral Salmonella-S-DNA-control boosted with i.p. S-peptide, oral Salmonella-tPA-S-DNA, oral Salmonella-tPA-S-DNA boosted with i.p S-peptide, oral Salmonella-CTLA4-S-DNA and oral Salmonella-CTLA4-S-DNA boosted with i.p. S-peptide showed neutralizing antibody titers of <1:20-1:160. Sera of all the mice immunized with i.m. tPA-S-DNA boosted with i.p. S-peptide and i.m. CTLA4-S-DNA boosted with i.p. S-peptide showed neutralizing antibody titers of >or=1:1280. The present observation may have major practical value, such as immunization of civet cats, since production of recombinant proteins from E. coli is far less expensive than production of recombinant proteins using eukaryotic systems.

show abstract

Section: Introductionmentioning

confidence: 51%

SARS coronavirus spike polypeptide DNA vaccine priming with recombinant spike polypeptide from Escherichia coli as booster induces high titer of neutralizing antibody against SARS coronavirus

Woo

Lau

Tsoi

et al. 2005

Vaccine

View full text Add to dashboard Cite

show abstract

“…Furthermore, the creation of scFvs fused to other protein is believed to lead to the formation of dimers [16], which should have greater avidity and stability than original monovalent scFvs, and can be used, for example, for detection purposes. At present, the laboratory tests that can be applied for routine diagnosis of SARS mainly include detecting viral genome by PCR involving in RT-PCR [19][20][21][22] and RT-LAMP [23], and detecting serum antibodies using SARS-CoV from Vero cell culture by ELI-SA [24][25][26], IFA [27][28][29], and NT [2]. However, there is little progress in detection of SARS-CoV using scFv-antibodies.…”

Section: Discussionmentioning

confidence: 99%

Identification of single-chain antibody fragments specific against SARS-associated coronavirus from phage-displayed antibody library

Liu

et al. 2005

Biochemical and Biophysical Research Communications

View full text Add to dashboard Cite

To develop early diagnostic reagents, effective vaccines, and even drugs against SARS-associated coronavirus (SARS-CoV), the human single fold single-chain antibody fragments, (scFv) libraries I+J (Tomlinson I+J) were used to identify novel scFvs, which can specifically bind to SARS-CoV. Interestingly, two scFvs (B5 and B9) exhibited higher binding specificity to SARS-CoV with the OD(450) value 0.608 and 0.545, respectively, and their coding sequences shared the identical sequence composed of V(H) gene (351bp) and V(L) gene (327bp), so the two scFvs were uniformly named as SA59B and chosen for further analysis. SA59B scFv was expressed in soluble form in Escherichia coli HB2151 and purified by immobilized metal affinity chromatography. The soluble 30kDa SA59B scFv-antibody was verified in SDS-PAGE and Western-blot. The purified SA59B scFv-antibody was labeled with HRP by the glutaraldehyde method, and the concentration of HRP and SA59B scFv-antibody in the SA59B-HRP solution reached 2.4 and 2.28mg/ml, respectively. Then, the binding ability of SA59B-HRP to SARS-CoV was evaluated by ELISA with S/N of 11.6, indicating higher binding specificity between them. Finally, both the SA59B sequence specificity and its application for diagnosis, prophylaxis or therapy of SARS were discussed.

show abstract

“…This assay provided a high degree of sensitivity (95.9%) for convalescent serum samples and specificity close to 90% [16]. Moreover, a peptide-ELISA has been used for retrospective serosurveillance of SARS [80][81]. This assay has been developed by epitope mapping, using synthetic peptides from the spike, the membrane and nucleocapsid protein sequences of SARS-CoV.…”

Section: A3 Hepatitis a Virus (Hav) Infectionmentioning

confidence: 98%

Synthetic Peptides for the Immunodiagnosis of Human Diseases

Gómara¹,

Haro

2007

CMC

View full text Add to dashboard Cite

Synthetic peptides have been shown to be valuable tools for viral laboratory diagnosis and can provide uniform, chemically well-defined antigens for antibody analysis, reducing inter- and intra-assay variation. The main aim in the development of peptide-based diagnostic tests is to recognise specific antibodies induced by the whole viral proteins but using selected short fragments containing the most potent antigenic determinants. The success of this approach depends on the extent to which synthetic peptides are able to mimic the immunodominant epitopes of antigens. In recent years, synthetic peptides that mimic specific epitopes of infectious agents' proteins have been used in diagnostic systems for various human diseases. The present review summarizes some of the drawbacks of the use of relatively short linear peptides as antigenic substrates and the subsequent chemical strategies developed in order to overcome the low peptide reactivity against specific antibodies. Moreover, it outlines the most significant bibliography published in the last five years which provides validated peptide based tests potentially useful for diagnosis of viral, bacterial, parasitic and autoimmune diseases.

show abstract

SARS Antibody Test for Serosurveillance

Abstract: A standardized and rapid peptide-based SARS assay is characterized for sensitivity and specificity.

Cited by 31 publications

References 20 publications

SARS coronavirus spike polypeptide DNA vaccine priming with recombinant spike polypeptide from Escherichia coli as booster induces high titer of neutralizing antibody against SARS coronavirus

SARS coronavirus spike polypeptide DNA vaccine priming with recombinant spike polypeptide from Escherichia coli as booster induces high titer of neutralizing antibody against SARS coronavirus

Identification of single-chain antibody fragments specific against SARS-associated coronavirus from phage-displayed antibody library

Synthetic Peptides for the Immunodiagnosis of Human Diseases

Contact Info

Product

Resources

About