A convenient immunochromatographic test strip for rapid detection of Scylla serrata reovirus.
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ABSTRACT:
SUBMITTER: Fan D
PROVIDER: S-EPMC6598930 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:A rapid, portable, and semi-quantitative immunochromatographic strip was developed for rapid and visual detection of alternariol monomethyl ether (AME). For this purpose, the anti-AME monoclonal antibody (mAb) was prepared and identified. AME coupled to bovine serum albumin (BSA) via methyl 4-bromobutanoate was prepared as immunogen. The recoveries of AME in spiked cherry and orange fruits determined by competitive ELISA were 86.1% and 80.7%, respectively. A colloidal gold nanoparticle (CGN) and CGNs-mAb conjugate were synthesized, and on this basis, a competitive colloidal gold immunochromatographic strip was developed and applied to the detection of AME toxin in fruit samples. The intensity of red density of the test line (T line) is inversely proportional to AME concentration in the range 0.1-10 ng/mL. The visual limit of detection (LOD) of AME was found to be about 10 ng/mL. The semi-quantitative detection can be completed in 10 min. Moreover, the immunochromatographic strip has lower cross-reactivity with AME analogues, and it has a good stability performance (following 3 months of storage). Hence, the colloidal gold immunochromatographic strip could be used as a semi-quantitative tool for the on-site, rapid, and visual detection of AME in fruit.
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Project description:BACKGROUND:Staphylococcus aureus is one of the most important microorganisms that causes various human diseases by secreting virulence factors known as staphylococcal super antigens (SAgs). Staphylococcal Enterotoxin B (SEB) is a bacterial antigen that is responsible for food poisoning in humans. Among SEB detection methods, a lateral flow device (LFD) is ideal for rapid immunochromatographic tests because it is easy to use, requires minimal time to produce results, and does not require personnel training. OBJECTIVES:In our laboratory, the production of an immunochromatographic test strip, for the detection of SEB using a sandwich assay and a competitive method, was described; the test can detect SEB with high sensitivity. MATERIALS AND METHODS:The strip assays were compared with PCR, a valid method for detection. For PCR, a specific sequence for SEB production was detected using primers designed according to GenBank sequences. RESULTS:In total, 80 food samples suspected of SEB contamination were assessed using the two methods. Fifty-four samples were contaminated based on the PCR technique and twenty-six of those were confirmed using the strip assay. CONCLUSIONS:The sensitivity of the sandwich method was approximately 10 ng/mL and that of the competitive method was approximately 250 ng/mL. In the LFD, a highly specific monoclonal antibody used for both the sandwich and competitive methods resulted in an increased sensitivity and accuracy for the detection of a minimal SEB concentration.
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Project description:A colloidal gold (ICS) test was developed for rapid detection of zearalenone (ZEN) in wheat samples. The mAb against ZEN was prepared in our laboratory and labelled with colloidal gold as a probe for the ICS test. The conditions were optimized and 30 nm colloidal gold nanoparticles were chosen for optimal performance. Millipore 135 was chosen as the NC membrane for its level of sensitivity. The optimum amount of coated antigen ZEN-OVA and anti-ZEN mAb was 0.5 mg/mL and 8 μg/mL, respectively. The ICS test, which has a detection limit of 15 ng/mL for ZEN, could be completed in 5 min. Analysis of ZEN in 202 wheat samples over three consecutive years revealed that data obtained from the ICS test were in a good agreement with LC-MS/MS data. This result demonstrated that the ICS test could be used as a qualitative tool to screen on-site for ZEN.
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Project description:In Uganda, bovine babesiosis continues to cause losses to the livestock industry because of shortages of cheap, quick, and reliable diagnostic tools to guide prescription measures. In this study, the presence of antibodies to Babesia bigemina and Babesia bovis in 401 bovine blood samples obtained from eastern and central areas of Uganda were detected using enzyme-linked immunosorbent assays (ELISAs) and immunochromatographic test strips (ICTs). The ELISA and ICT test used targeted the B. bigemina C-terminal rhoptry-associated protein (RAP-1/CT17) and B. bovis spherical body protein-4 (SPB-4). Using ELISA, single-ICT and dual-ICT, positive samples for B. bovis were detected in 25 (6.2%), 17 (4.3%), and 14 (3.7%) samples respectively, and positive samples for B. bigemina were detected in 34 (8.4%), 27 (6.7%), and 25 (6.2%), respectively. Additionally, a total of 13 animals (3.2%) had a mixed infection. The correlation between ELISA and single-ICT strips results revealed slight agreement with kappa values ranging from 0.088 to 0.191 between both methods, while the comparison between dual-ICT and single-ICT results showed very good agreement with kappa values >0.80. This study documented the seroprevalence of bovine babesiosis in central and eastern Uganda, and showed that ICT could, after further optimization, be a useful rapid diagnostic test for the diagnosis of bovine babesiosis in field settings.
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