Protein A-Based ELISA: Its Evaluation in the Diagnosis of Herpes Simplex Encephalitis

Bhullar, Shradha S.; Kashyap, Rajpal S.; Chandak, Nitin H.; Purohit, Hemant J.; Taori, Girdhar M.; Daginawala, Hatim F.

doi:10.1089/vim.2010.0129

Cited by 6 publications

(3 citation statements)

References 23 publications

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“…Additionally, SpG can bind to Fab in the first constant region of the Ig γ chain (CH1γ)13. Because of their unique antibody binding features, IBPs have fundamental applications in biological and medical sciences, such as antibody purification, antibody diagnostic detection, immunoadsorption therapy, and immunoprecipitation assays14151617.…”

mentioning

confidence: 99%

In vitro molecular evolution yields an NEIBM with a potential novel IgG binding property

Ding

et al. 2014

Sci Rep

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Staphylococcus aureus protein A (SpA) and protein G of groups C and G streptococci (SpG) are two well-defined bacterial immunoglobulin (Ig)-binding proteins (IBPs) with high affinity for specific sites on IgG from mammalian hosts. Both SpA and SpG contain several highly-homologous IgG-binding domains, each of which possesses similar binding characteristic of the whole corresponding proteins. Whether specific combinations of these domains could generate a molecule with novel IgG-binding properties remained unknown. We constructed a combinatorial phage library displaying randomly-rearranged A, B, C, D and E domains of SpA as well as the B2 (G2) and B3 (G3) domains of SpG. In vitro molecular evolution directed by human, rabbit, bovine, or goat polyclonal IgGs and four subclasses of mouse monoclonal IgGs generated one common combination, D-C-G3. A series of assays demonstrated that D-C-G3 exhibited a potential novel IgG binding property that was obviously different from those of both parent proteins. This study provides an example of successful protein engineering through in vitro molecular evolution and useful approaches for structure and function studies of IBPs.

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mentioning

confidence: 99%

In vitro molecular evolution yields an NEIBM with a potential novel IgG binding property

Ding

et al. 2014

Sci Rep

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“…As an alternative to enzyme-conjugated antibodies, the S. aureus protein A (PA) and streptococcal protein G (PG) have been applied in the diagnosis of many infectious diseases with the advantage of having an affinity for immunoglobulins from various animal species. The evidence for a higher binding affinity of both PA and PG to IgG, promoting the detection of lower amounts of antibodies than anti-IgG conjugates, has been demonstrated in the diagnosis of diverse infectious diseases [ 29 , 30 ].…”

Section: Discussionmentioning

confidence: 99%

Refinement of the rKLi8.3-Based Serodiagnostic ELISA Allows Detection of Canine Leishmaniosis in Dogs with Low Antibody Titers

Teixeira,

Valle,

Mahdavi

et al. 2024

Pathogens

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The diagnosis of canine leishmaniasis (CanL) still represents a challenge due to the variable clinical manifestations and the large number of asymptomatic dogs. Serological tests are most commonly used to detect infected animals, revealing anti-Leishmania antibodies, mainly of the IgG isotype. Recently, a new diagnostic antigen, rKLi8.3, containing 8.3 kinesin tandem repeats (TR) from a Leishmania infantum strain from Sudan, has been shown to provide excellent specificity and sensitivity for the detection of Leishmania-infected humans and dogs. However, asymptomatic animals with very low antibody titers are often difficult to detect by serodiagnosis. Thus, we wondered whether the addition of an anti-IgG-enhancing step in the protein A/G-based rKLi8.3-ELISA will improve the diagnostic performance without decreasing the specificity. For this, parasitologically confirmed CanL cases with low or high clinical scores, uninfected healthy controls and dogs with other infections were tested by rKLi8.3-ELISA as well as two different immunochromatographic rapid tests, rKLi8.3-lateral flow test (LFT) and Dual Path Platform (DPP®) based on the rK28 antigen. Our results show that the diagnostic accuracies of the rKLi8.3-ELISA and LFT were similar to that of DPP, missing several asymptomatic animals. However, the addition of a secondary, amplifying anti-dog IgG antibody in the protein A/G-based rKLi8.3-ELISA enabled the detection of nearly all asymptomatic dogs without compromising its specificity.

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“…The early diagnosis of HSE is difficult as the clinical picture of encephalitis caused by HSV is similar to that of other viral infections (Bhullar et al, 2011). PCR is although recognized as the standard laboratory technique for HSE diagnosis, however, it remains negative in cases where CSF has been obtained at late times following onset of illness or when the viral load is low in CSF.…”

Section: Discussionmentioning

confidence: 99%

Identification of an immunodominant epitope in glycoproteins B and G of herpes simplex viruses (HSVs) using synthetic peptides as antigens in assay of antibodies to HSV in herpes simplex encephalitis patients

Ss¹,

Nh²,

Nn³

et al. 2014

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Summary. -Herpes simplex encephalitis (HSE) is a severe viral infection of the central nervous system (CNS). Assay of antibody response is widely used in diagnostics of HSE. The aim of this study was to identify an immunodominant epitope determining the antibody response to herpes simplex viruses (HSVs) in cerebrospinal fluid (CSF) of HSE patients. The synthetic peptides that resembled type-common as well as type-specific domains of glycoproteins B (gB) and G (gG) of these viruses were evaluated for binding with IgM and IgG antibodies in CSF samples from HSE and non-HSE patients in ELISA. The QLHDLRF peptide, derived from gB of HSV was found to be an immunodominant epitope in the IgM and IgG antibody response. The patients with confirmed and suspected HSE showed in ELISA against this peptide 26% and 23% positivities for IgM, 43% and 37% positivities for IgG and 17% and 15% for both IgM and IgG antibodies, respectively. The total positivities of 86% and 75% for both IgM and IgG antibodies were obtained in the patients with confirmed and suspected HSE, respectively. These results demonstrate that a synthetic peptide-based diagnostics of HSE can be an efficient and easily accessible alternative. This is the first report describing the use of synthetic peptides derived from HSVs in diagnostics of HSE using patientsʹ CSF samples.

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Protein A-Based ELISA: Its Evaluation in the Diagnosis of Herpes Simplex Encephalitis

Cited by 6 publications

References 23 publications

In vitro molecular evolution yields an NEIBM with a potential novel IgG binding property

In vitro molecular evolution yields an NEIBM with a potential novel IgG binding property

Refinement of the rKLi8.3-Based Serodiagnostic ELISA Allows Detection of Canine Leishmaniosis in Dogs with Low Antibody Titers

Identification of an immunodominant epitope in glycoproteins B and G of herpes simplex viruses (HSVs) using synthetic peptides as antigens in assay of antibodies to HSV in herpes simplex encephalitis patients

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