Protein-G-based lateral flow assay for rapid serodiagnosis of brucellosis in domesticated animals

Manasa, M; Revathi, P; Chand, Mohammed; Maroudam, V.; Navaneetha, P.; Raj, G. Dhinakar; Kishor, P. B. Kavi; De, Barun K.; Polavarapu, Rathnagiri

doi:10.1080/15321819.2018.1541803

Cited by 14 publications

(12 citation statements)

References 17 publications

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“…These procedures account for the optimal performance of the iELISA used here [3, 14] and, therefore, caution has to be taken to draw conclusions from LFA studies compared with standardized iELISA in a different way [10–13] or with competitive ELISA [8]. The iELISA used in previous works applied cutoffs recommended by manufacturers without documented support [12] or established using the mean and standard deviation of the optical density of the populations included and/or sera of animals of unknown brucellosis status [10, 11, 13], methods that do not allow to set proper diagnostic cutoffs [14].…”

Section: Discussionmentioning

confidence: 99%

“…Shome et al [10] also found parallelism between RBT and an in-house developed bovine-LFA in the sera of 153 buffaloes of unknown individual brucellosis status using an indirect ELISA (iELISA) as the reference. Manasa et al [11] investigated a protein G-based LFA using sera of cattle (including buffaloes), small ruminants and pigs of unknown brucellosis status and also reported similar results for RBT and LFA. On the other hand, discrepancies between RBT and bovine-LFA and low relative specificity or sensitivity, respectively, have been reported in two studies with sera (n = 40 in both) of cattle of unknown brucellosis status using an iELISA as the reference [12, 13].…”

Section: Introductionmentioning

confidence: 86%

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Comparative performance of lateral flow immunochromatography, iELISA and Rose Bengal tests for the diagnosis of cattle, sheep, goat and swine brucellosis

et al. 2019

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Background Brucellosis is a world-wide extended zoonosis that causes a grave problem in developing economies. Animal vaccination and diagnosis are essential to control brucellosis, and the need for accurate but also simple and low-cost tests that can be implemented in low-infrastructure laboratories has been emphasized. Methodology We evaluated bovine, sheep, goat and swine lateral flow immunochromatography assay kits (LFA), the Rose Bengal test (RBT) and a well-validated protein G indirect ELISA (iELISA) using sera of Brucella culture-positive and unvaccinated brucellosis free livestock. Sera from cattle vaccinated with S19 and RB51 brucellosis vaccines were also tested. Finally, we compared RBT and LFA using sera of white Fulani cattle of unknown bacteriological status from a brucellosis endemic area of Nigeria. Results and conclusions Although differences were not statistically significant, RBT showed the highest values for diagnostic sensitivity/specificity in cattle (LFA, 96.6/98.8; RBT, 98.9/100; and iELISA, 96.6/100) and the iELISA yielded highest values in sheep (LFA, 94.0/100; RBT, 92.0/100; iELISA, 100/100), goats (LFA, 95.7/96.2; RBT, 97.8/100; iELISA, 100/100) and pigs (LFA, 92.3/100; RBT, 92.3/100; iELISA, 100/100). Vaccine S19 administered subcutaneously interfered in all tests but conjunctival application minimized the problem. Although designed not to interfere in serodiagnosis, vaccine RB51 interfered in LFA and iELISA but not in the RBT. We found closely similar apparent prevalence results when testing the Nigerian Fulani cattle by RBT and LFA. Although both RBT and LFA (showing similar diagnostic performance) are suitable for small laboratories in resource-limited areas, RBT has the advantage that a single reagent is useful in all animal species. Considering these advantages, its low cost and that it is also useful for human brucellosis diagnosis, RBT might be a good choice for resource-limited laboratories.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 86%

Comparative performance of lateral flow immunochromatography, iELISA and Rose Bengal tests for the diagnosis of cattle, sheep, goat and swine brucellosis

et al. 2019

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“…To confirm the flexibility of the two devices, both were used to test sera from dogs [4] and cats [14]. The results are reported in Table 3.…”

Section: Resultsmentioning

confidence: 99%

“…In a previous work, we demonstrated the high sensitivity and specificity reached by using the total antibody approach [13]. In other studies, the effects of using Staphylococcal protein A (SpA) and Streptococcal protein G (SpG) as detection [14,15] or capturing elements [16] on antibody testing have been explored. Another interesting benefit of bacterial bioligands is their broad selectivity towards immunoglobulins of different animal species.…”

Section: Introductionmentioning

confidence: 98%

Bacterial ligands as flexible and sensitive detectors in rapid tests for antibodies to SARS-CoV-2

et al. 2022

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Lateral flow immunoassay (LFIA) is widely employed as point-of-care tests (POCT) for the diagnosis of infectious diseases. The accuracy of LFIA largely depends on the quality of the immunoreagents used. Typical LFIAs to reveal the immune response to severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) employ anti-human immunoglobulin (hIG) antibodies and recombinant viral antigens, which usually are unstable and poorly soluble. Broad selective bacterial proteins, such as Staphylococcal protein A (SpA) and Streptococcal protein G (SpG) can be considered alternatives to anti-hIG to increase versatility and sensitivity of serological LFIAs because of their high binding capacity, interspecies reactivity, and robustness. We developed two colorimetric LFA devices including SpA and SpG linked to gold nanoparticles (GNP) as detectors and explored the use of a specific, stable, and soluble immunodominant fraction of the nucleocapsid protein from SARS-CoV-2 as the capturing agent. The optimal amount of SpA-GNP and SpG-GNP conjugates and the protein-to-GNP ratios were defined through a full factorial experimental design to maximize the diagnostic sensitivity of the LFIAs. The new LFA devices were applied to analyze 105 human serum samples (69 positive and 36 negatives according to reference molecular diagnostic methods). The results showed higher sensitivity (89.9%, 95% CI 82.7–97.0) and selectivity (91.7%, 82.6–100) for the SpA-based compared to the SpG-based LFA. In addition, 18 serum samples from cats and dogs living with COVID-19 patients were analyzed and 14 showed detectable levels of anti-SARS-CoV-2 antibodies, thus illustrating the flexibility of the SpA- and SpG-based LFAs. Graphical abstract

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“…In the present study, the composition of GNP conjugates with streptococcal protein G was investigated using the previously developed technique based on fluorescence spectroscopy [22]. Streptococcal protein G was chosen for the study because of its wide immunoanalytical use [23][24][25]. The binding ability of the obtained conjugates in immunochromatographic assay (ICA), namely in the determination of specific antibodies against Brucella abortus lipopolysaccharide (LPS), was studied.…”

Section: Introductionmentioning

confidence: 99%

Correlation between composition and activity of gold nanoparticle conjugates with streptococcal protein G

2020

Biointerface Res Appl Chem

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The composition of the conjugates of gold nanoparticles with streptococcal protein G was studied using fluorescence spectroscopy. The method for determining the composition is based on measuring the intrinsic fluorescence of tryptophan as part of the protein. The equilibrium constants of protein binding by the gold surface were determined using the Sketchard method. An increase in the dissociation constant of the protein–nanoparticle complex for increasing the amount of bound protein was demonstrated, and a relationship was established between the stability of the conjugates, their antigen-binding activity, and the dissociation constant. The effectiveness of the conjugates of different compositions in immunochromatographic assay of specific antibodies against the lipopolysaccharide antigen of Brucella abortus was compared. The binding ability of the conjugates increased along with the amount of protein G to ~200 molecules per nanoparticle. A further increase in the amount of adsorbed protein led to a deterioration in the functional activity of the conjugates.

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Protein-G-based lateral flow assay for rapid serodiagnosis of brucellosis in domesticated animals

Cited by 14 publications

References 17 publications

Comparative performance of lateral flow immunochromatography, iELISA and Rose Bengal tests for the diagnosis of cattle, sheep, goat and swine brucellosis

Comparative performance of lateral flow immunochromatography, iELISA and Rose Bengal tests for the diagnosis of cattle, sheep, goat and swine brucellosis

Bacterial ligands as flexible and sensitive detectors in rapid tests for antibodies to SARS-CoV-2

Correlation between composition and activity of gold nanoparticle conjugates with streptococcal protein G

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