scholarly journals Development of a Scrub Typhus Diagnostic Platform Incorporating Cell-Surface Display Technology

2021 ◽  
Vol 12 ◽  
Author(s):  
Chih-Chi Liao ◽  
Chih-Hsuan Tsai ◽  
Huei-Ru Lo ◽  
Pey-Ru Lin ◽  
Chang-Chi Lin ◽  
...  
Keyword(s):  
Scrub Typhus ◽  
Low Cost ◽  
Surface Display ◽  
Cell Surface Display ◽  
Asia Pacific ◽  
Detection Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Passenger Domain ◽  
Display Technology

Scrub typhus (ST), also known as tsutsugamushi disease and caused by rickettsia Orientia tsutsugamushi, is an underestimated fatal epidemic in the Asia-Pacific region, resulting in a million human infections each year. ST is easily misdiagnosed as clinical diagnosis is based on non-specific skin eschar and flu-like symptoms. Thus, the lack of accurate, convenient, and low-cost detection methods for ST poses a global health threat. To address this problem, we adopted baculovirus surface-display technology to express three variants of TSA56, the major membrane antigen of O. tsutsugamushi, as well as the passenger domain of ScaC (ScaC-PD), on insect Sf21 cell surfaces rather than biosafety level 3 bacteria in an enzyme-linked immunosorbent assay (ELISA). Recombinant TSA56 and ScaC-PD were all properly expressed and displayed on Sf21 cells. Our cell-based ELISA comprising the four antigen-displaying cell types interacted with monoclonal antibodies as well as serum samples from ST-positive field-caught rats. This cell-based ELISA presented high accuracy (96.3%), sensitivity (98.6%), and specificity (84.6%) when tested against the ST-positive rat sera. Results of a pilot study using human sera were also highly consistent with the results of immunofluorescence analyses. By adopting this approach, we circumvented complex purification and refolding processes required to generate recombinant O. tsutsugamushi antigens and reduced the need for expensive equipment and extensively trained operators. Thus, our system has the potential to become a widely used serological platform for diagnosing ST.

scholarly journals Corrigendum: Development of a Scrub Typhus Diagnostic Platform Incorporating Cell-Surface Display Technology

2021 ◽  
Vol 12 ◽  
Author(s):  
Chih-Chi Liao ◽  
Chih-Hsuan Tsai ◽  
Huei-Ru Lo ◽  
Pey-Ru Lin ◽  
Chang-Chi Lin ◽  
...  
Keyword(s):  
Cell Surface ◽  
Scrub Typhus ◽  
Surface Display ◽  
Cell Surface Display ◽  
Display Technology

scholarly journals Standardization of dot-ELISA for the serological diagnosis of toxocariasis and comparision of the assay with ELISA

1992 ◽  
Vol 34 (1) ◽  
pp. 55-60 ◽  
Author(s):  
Eide Dias Camargo ◽  
Paulo Mutuko Nakamura ◽  
Adelaide José Vaz ◽  
Marcos Vinícius da Silva ◽  
Pedro Paulo Chieffi ◽  
...  
Keyword(s):  
Low Cost ◽  
Clinical Signs ◽  
Toxocara Canis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Specific Antibodies ◽  
Normal Individuals ◽  
Before And After ◽  
Dot Elisa ◽  
Stability Time

The dot-enzyme-linked immunosorbent assay (dot-ELISA) was standardized using somatic (S) and excretory-secretory (ES) antigens of Toxocara-canis for the detection of specific antibodies in 22 serum samples from children aged 1 to 15 years, with clinical signs of toxocariasis. Fourteen serum samples from apparently normal individuals and 28 sera from patients with other pathologies were used as controls. All samples were used before and after absorption with Ascaris suum extract. When the results were evaluated in comparison with ELISA, the two tests were found to have similar sensitivity, but dot-ELISA was found to be more specific in the presence of the two antigens studied. Dot-ELISA proved to be effective for the diagnosis of human toxocariasis, presenting advantages in terms of yield, stability, time and ease of execution and low cost.

Cell-surface display technology and metabolic engineering of Saccharomyces cerevisiae for enhancing xylitol production from woody biomass

2019 ◽  
Vol 21 (7) ◽  
pp. 1795-1808 ◽  
Author(s):  
Gregory Guirimand ◽  
Kentaro Inokuma ◽  
Takahiro Bamba ◽  
Mami Matsuda ◽  
Kenta Morita ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Metabolic Engineering ◽  
Cell Surface ◽  
Surface Display ◽  
Woody Biomass ◽  
Cell Surface Display ◽  
Xylitol Production ◽  
Display Technology ◽  
Pharmaceutical Industries

Xylitol is a major commodity chemical widely used in both the food and pharmaceutical industries.

scholarly journals Scrub typhus: a hospital-based study in the northern districts of West Bengal, India

2019 ◽  
Vol 7 (6) ◽  
pp. 2403
Author(s):  
Sanjay Kumar Mallick ◽  
Santanu Hazra ◽  
Tanmoy Nandi ◽  
Arunabha Sarkar
Keyword(s):  
Case Fatality Rate ◽  
West Bengal ◽  
Scrub Typhus ◽  
Case Fatality ◽  
Febrile Illness ◽  
Asia Pacific ◽  
Enzyme Linked Immunosorbent Assay ◽  
Acute Febrile Illness ◽  
Fatality Rate ◽  
Serological Tests

Background: Scrub typhus caused by Orientia tsutsugamushi, is a mite-borne zoonotic acute febrile illness. Geographically, it is confined to the Asia-Pacific region and important re-emerging infection in India. Clinical diagnosis of scrub typhus from other acute febrile illness is very difficult due to nonspecific symptoms and the relative absence of eschar in the Indian population. Case fatality rate varies from 30-70% depending on the clinical suspicion, delay in diagnosis and treatment. Antibody-based serological tests are the mainstay of diagnosis. IgM enzyme-linked immunosorbent assay (ELISA) against O. tsutsugamushi is helpful for the diagnosis of scrub typhus within the first week of illness.Methods: The aim of the study was to determine the prevalence of the disease in Northern districts of West Bengal, India using IgM ELISA.Results: Out of 577 serum samples tested 10.05% were positive for IgM antibodies. Majority of cases were below 40 years of age with higher prevalence in female patients. The disease showed a seasonal trend with a peak during the monsoon and later months. The case fatality rate among ELISA positive cases was 32.76%.Conclusions: Significant seropositivity against scrub typhus among cases of acute febrile illness with relatively higher mortality indicates that scrub typhus should be included in the differential diagnosis and confirmed by IgM ELISA.

scholarly journals Detection of C-Reactive Protein Using Histag-HRP Functionalized Nanoconjugate with Signal Amplified Immunoassay

Nanomaterials ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 1240
Author(s):  
Mohd Farhan Siddiqui ◽  
Zeeshan Ahmad Khan ◽  
Seungkyung Park
Keyword(s):  
Detection Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
C Reactive Protein ◽  
Serum Samples ◽  
Reactive Protein ◽  
Elisa Format ◽  
Early Biomarker ◽  
Cost Efficient ◽  
Detection Of Biomarkers ◽  
Gold Nanoprobe

Ultrasensitive detection of biomarkers is highly significant for disease prognosis and public health treatment. Despite wide acceptance in routine laboratory tests, the conventional enzyme-linked immunosorbent assay (ELISA) has been of limited use for early biomarker detection due to insufficient sensitivity and multiple long incubation time. Several nanoprobes have been introduced to circumvent the limitation, however, rapid, simple, and chemical-free nanoprobe synthesis and sensitive detection methods, particularly for ELISA, are still lacking. In this study, we have synthesized a gold nanoprobe, conjugated with multiple 6X-histidine (6X-his) peptide and nickel-horseradish peroxidase (Ni2+-HRP), for enhancing the colorimetric signal in ELISA. The developed nanoprobe has been tested for the detection of immunologically significant C-reactive protein (CRP) in ELISA format. The performance of designed probe is validated by testing standard and serum samples, and the detection limit of 32.0 pg/mL with R2 = 0.98 is confirmed. Furthermore, a comparative analysis of the developed nanoprobe was performed with ELISA developed on conventional guidelines, the proposed immunoassay showed an increase of 12-fold sensitivity for detecting CRP due to the high loading of 6Xhis peptide and binding of multiple Ni2+-HRP on a gold nanoparticle. Additionally, the proposed assay provides a simple, fast, and cost-efficient (not requiring multiple antibodies) detection of CRP with easy nanoprobe synthesis. Moreover, the developed Histag-HRP functionalized nanoconjugate immunoassay is flexible and can be applied to other biomarkers efficiently by using disease specific antibody.

scholarly journals Surface Display Technology for Biosensor Applications: A Review

Sensors ◽  
2020 ◽  
Vol 20 (10) ◽  
pp. 2775 ◽  
Author(s):  
Min Park
Keyword(s):  
Mammalian Cells ◽  
Surface Display ◽  
Cell Surface Display ◽  
Bacterial Surface ◽  
Yeast Cell Surface ◽  
Bacterial Surface Display ◽  
Display Technology ◽  
Almost All ◽  
Display Systems ◽  
Biosensor Applications

Surface display is a recombinant technology that expresses target proteins on cell membranes and can be applied to almost all types of biological entities from viruses to mammalian cells. This technique has been used for various biotechnical and biomedical applications such as drug screening, biocatalysts, library screening, quantitative assays, and biosensors. In this review, the use of surface display technology in biosensor applications is discussed. In detail, phage display, bacterial surface display of Gram-negative and Gram-positive bacteria, and eukaryotic yeast cell surface display systems are presented. The review describes the advantages of surface display systems for biosensor applications and summarizes the applications of surface displays to biosensors.

scholarly journals Evaluation of an Immunoglobulin M Capture Enzyme-Linked Immunosorbent Assay for Diagnosis of Orientia tsutsugamushi Infection

2003 ◽  
Vol 10 (3) ◽  
pp. 394-398 ◽  
Author(s):  
Won-Jong Jang ◽  
Myung-Suk Huh ◽  
Kyung-Hee Park ◽  
Myung-Sik Choi ◽  
Ik-Sang Kim
Keyword(s):  
Immunosorbent Assay ◽  
Scrub Typhus ◽  
Microtiter Plate ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Orientia Tsutsugamushi ◽  
Serum Samples ◽  
Capture Elisa ◽  
Igm Antibodies ◽  
Immunodominant Antigen

ABSTRACT To differentiate scrub typhus from other acute febrile diseases, a rapid and reliable serological diagnosis is important. We developed an immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (ELISA) for diagnosis of recent Orientia tsutsugamushi infections in humans. The 56-kDa major outer membrane protein of O. tsutsugamushi is well known as the most immunodominant antigen in scrub typhus. The test is based on the use of the biotinylated recombinant 56-kDa protein of O. tsutsugamushi Boryong, Bor56, which was expressed as a fusion protein with a maltose-binding protein in Escherichia coli. In the test, the serum IgM antibodies were captured by anti-human IgM antibodies coated onto a microtiter plate. The captured IgM antibodies were revealed through sequential addition of biotinylated Bor56 antigen and peroxidase-conjugated streptavidin to the plate. The IgM capture ELISA was compared with the immunofluorescence antibody assay (IFA) by testing 176 serum samples from patients with diagnosed cases of rickettsial disease and patients with other acute febrile diseases. Of the 81 IgG IFA-positive samples, 78 tested positive (sensitivity, 96.3%) and all 31 IgM IFA-positive samples tested positive (sensitivity, 100%) by the IgM capture ELISA. The specificity of the IgM capture ELISA was 99%, and 1 of the 95 IFA-negative samples was positive in the assay. These results strongly suggest that IgM capture ELISA using the recombinant Bor56 antigen is a reliable and detailed method for the detection of early O. tsutsugamushi infection.

An ELISA kit with two detection modes for the diagnosis of lymphatic filariasis

2014 ◽  
Vol 89 (5) ◽  
pp. 552-558 ◽  
Author(s):  
S. Wongkamchai ◽  
W. Satimai ◽  
S. Loymek ◽  
H. Nochot ◽  
J.J. Boitano
Keyword(s):  
Lymphatic Filariasis ◽  
Low Cost ◽  
Kappa Statistic ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Laboratory Equipment ◽  
Elisa Kit ◽  
Naked Eye ◽  
Test Kit ◽  
Endemic Areas

AbstractThe aim of this study was to develop a low-cost antifilarial immunoglobulin (Ig) G4 detection kit for the diagnosis of lymphatic filariasis. The kit was designed to be used by minimally trained personnel without the constraints of expensive laboratory equipment. We provide a description of the development and validation of a single-serum-dilution based enzyme-linked immunosorbent assay (ELISA) kit with ready-to-use reagents for measuring antifilarial IgG4 antibodies. The kit was tested on residents in Brugia malayi-endemic areas in southern Thailand. Detection was performed by naked-eye observation of the resultant colour of the immunological reactivity. The coefficient of variation (CV) was used to assess the reproducibility of the results. Long-term stability was measured over a 6-month period. Sensitivity of the test kit was 97% when compared with microfilariae detection in thick blood smears. Specificity was 98.7% based on the sera of 57 patients living outside the endemic areas who were infected with other parasites and 100 parasite-free subjects. All positive CVs were < 10%. The test kit was remarkably stable over 6 months. Field validation was performed by the detection of antifilarial IgG4 in 4365 serum samples collected from residents of brugian filariasis-endemic areas and compared with outcome colours of the test samples by the naked eye. Subsequent ELISA evaluation of these results using an ELISA reader indicated high agreement by the kappa statistic. These results demonstrate that the test kit is efficient and useful for public health laboratories as an alternative tool for the diagnosis of lymphatic filarial infection.

scholarly journals Progress in Bacillus subtilis Spore Surface Display Technology towards Environment, Vaccine Development, and Biocatalysis

2017 ◽  
Vol 27 (3) ◽  
pp. 159-167 ◽  
Author(s):  
Huayou Chen ◽  
Jawad Ullah ◽  
Jinru Jia
Keyword(s):  
Bacillus Subtilis ◽  
Industrial Development ◽  
Vaccine Development ◽  
Low Cost ◽  
Surface Display ◽  
Cell Surface Display ◽  
Coat Proteins ◽  
Spore Surface ◽  
Spore Surface Display ◽  
Alkaline Tolerance

Spore surface display is the most desirable with enhanced effects, low cost, less time consuming and the most promising technology for environmental, medical, and industrial development. Spores have various applications in industry due to their ability to survive in harsh industrial processes including heat resistance, alkaline tolerance, chemical tolerance, easy recovery, and reusability. Yeast and bacteria, including gram-positive and -negative, are the most frequently used organisms for the display of various proteins (eukaryotic and prokaryotic), but unlike spores, they can rupture easily due to nutritive properties, susceptibility to heat, pH, and chemicals. Hence, spores are the best choice to avoid these problems, and they have various applications over nonspore formers due to amenability for laboratory purposes. Various strains of <i>Clostridium</i> and <i>Bacillus</i> are spore formers, but the most suitable choice for display is <i>Bacillus subtilis</i> because, according to the WHO, it is safe to humans and considered as “GRAS” (generally recognized as safe). This review focuses on the application of spore surface display towards industries, vaccine development, the environment, and peptide library construction, with cell surface display for enhanced protein expression and high enzymatic activity. Different vectors, coat proteins, and statistical analyses can be used for linker selection to obtain greater expression and high activity of the displayed protein.

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