Detection Of IgG Research Articles

Dot Immunobinding Assay for the Rapid Serodetection of Scedosporium/Lomentospora in Cystic Fibrosis Patients.

The detection of Scedosporium/Lomentospora is still based on non-standardized low-sensitivity culture procedures. This fact is particularly worrying in patients with cystic fibrosis (CF), where these fungi are the second most common filamentous fungi isolated, because a poor and delayed diagnosis can worsen the prognosis of the disease. To contribute to the discovery of new diagnostic strategies, a rapid serological dot immunobinding assay (DIA) that allows the detection of serum IgG against Scedosporium/Lomentospora in less than 15 min was developed. A crude protein extract from the conidia and hyphae of Scedosporium boydii was employed as a fungal antigen. The DIA was evaluated using 303 CF serum samples (162 patients) grouped according to the detection of Scedosporium/Lomentospora in the respiratory sample by culture, obtaining a sensitivity and specificity of 90.48% and 79.30%, respectively; positive and negative predictive values of 54.81% and 96.77%, and an efficiency of 81.72%. The clinical factors associated with the results were also studied using a univariate and a multivariate analysis, which showed that Scedosporium/Lomentospora positive sputum, elevated anti-Aspergillus serum IgG and chronic Pseudomonas aeruginosa infection were significantly associated with a positive result in DIA, while Staphylococcus aureus positive sputum showed a negative association. In conclusion, the test developed can offer a complementary, rapid, simple and sensitive method to contribute to the diagnosis of Scedosporium/Lomentospora in patients with CF.

Rapid, sensitive, and specific lateral-flow immunochromatographic point-of-care device for detection of varicella-zoster virus immunoglobulin G antibodies in fingerstick blood

Varicella zoster virus (VZV) causes childhood chickenpox, becomes latent in sensory ganglia and reactivates years later to cause shingles (Zoster) and postherpetic neuralgia in the elderly and immunosuppressed individuals. Serologic IgG tests can be used to determine if a person has antibodies to VZV from past varicella infection or had received varicella or zoster (shingles) vaccination. Commercial enzyme-linked immunosorbent assays (ELISAs) are currently used for the detection of VZV IgG antibodies in patient serum samples. However, ELISA tests require collection and processing of blood samples in a CLIA laboratory to separate serum or plasma for further testing. In this paper, we describe the development and testing of an antibody based Lateral Flow Immunochromatographic assay (LFA) device for the detection of VZV IgG in fingerstick whole blood. Analytical and clinical analyses were performed to compare the performance characteristics of the Viro VZV IgG LFA (VZV LFA) and the Diamedix VZV IgG ELISA. Analytical studies demonstrated the higher sensitivity of the VZV LFA compared to the ELISA by testing dilutions of the WHO VZV IgG serum International Standard. Clinical performance characteristics of the VZV LFA fingerstick whole blood assay were assessed at three point of care (POC) facilities by untrained users testing samples from 300 prospectively enrolled study subjects. VZV LFA results were compared with results obtained by testing serum samples obtained from the same study participants by the Diamedix VZV IgG ELISA. Two specimens with invalid results by the LFA assay were not included in the LFA performance calculations and nine equivocal ELISA results were included as positive for IgG results. The results from all three POC clinical sites demonstrated the higher sensitivity/positive percent agreement (PPA) (99.26%, 95% CI: 97.34–99.80) of the VZV LFA compared to the Diamedix VZV IgG ELISA (94.08%, 95% CI: 90.72–96.27). The specificity/negative percent agreement (NPA) of the VZV LFA compared to the ELISA test was calculated initially to be 39.29% (95% CI: 23.57–57.59) with 19 discordant test results out of 298 test results between the two assays (17 LFA positive/ELISA negative and two LFA negative/ELISA positive). The PPA and true NPA of the VZV LFA were determined by testing all 298 samples, including the discordant (19) and all concordant negative and positive (279) study subject serum samples, before and after blocking VZV gE antibody sites in the samples by spiking with VZV LFA gE capture antigen. The NPA improved to 100% (95% CI: 74.12–100) after the procedure when compared to the ELISA test results. The comparator ELISA PPA based on the spiking/blocking study remained as 94.08%, (95% CI: 90.72–96.27), comparable to test results from untreated samples.The VZV LFA has been demonstrated to be simple and sufficiently robust for use in CLIA-waived POC facilities by untrained healthcare professionals and to detect VZV IgG in 20 min from fingerstick whole blood. The VZV LFA therefore provides a fast, reliable, and highly sensitive method of determining prior VZV viral infection or varicella and zoster vaccination status.

SEROPREVALENCE OF Toxoplasma go¬ndii IgG ANTIBODY IN WOMEN SUFFERING FROM ABORTION AT SANA’A CAPITAL AND CAPITAL TRUSTEESHIP

Toxoplasma gondii, is obligate intracellular protozoan, infects humans and any other warm-blooded animal. Toxoplasmosis is probably one of the most common human infections and becomes a life threatening disease in case of congenital infection or immunodepressed patient. The aim of this study was to estimate toxoplasmosis prevalence among Sana’a capital and capital trusteeship women suffering from abortion and to emphasize the consequences. The study was performed between 2006 and 2007 including women suffering from abortion. Latex agglutination (LAT) was used for Toxoplasma IgG detection. The result indicated that of 508 women suffering from abortion, 344 (67.7 %) were seropositive for Toxoplasmosis. The age (21-30 years) and (31-40 years) had the highest percentage (68.4 %) and (68.5 %) respectively followed by age (≤ 20 years) (66.0 %) of positive results. The most prevalent titers of LAT were 1/16 (25.0 %) followed by 1/32 (19.10 %) then 1/8 (8.30 %). The results confirmed Toxoplasma gondii wide spread at Sana’a capital and capital trusteeship and the risk represented by this protozoan. Therefore any raiser for this disease should be taken into consideration in order to diminish the consequences of this infection.

Antibody response, neutralizing potency, and transplacental antibody transfer following SARS-CoV-2 infection versus mRNA-1273, BNT162b2 COVID-19 vaccination in pregnancy.

To improve our understanding of the immune response, including the neutralization antibody response, following COVID-19 vaccination in pregnancy. This was a prospective cohort study comprising patients with PCR-confirmed SARS-CoV-2 infection and patients who received both doses of mRNA COVID-19 vaccine (mRNA-1273, BNT162b2) in pregnancy recruited from two hospitals in Atlanta, GA, USA. Maternal blood and cord blood at delivery were assayed for anti-receptor binding domain (RBD) IgG, IgA and IgM, and neutralizing antibody. The detection of antibodies, titers, and maternal to fetal transfer ratios were compared. Nearly all patients had detectable RBD-binding IgG in maternal and cord samples. The vaccinated versus infected cohort had a significantly greater proportion of cord samples with detectable neutralizing antibody (94% vs. 28%, P< 0.001) and significantly higher transfer ratios for RBD-specific IgG and neutralizing antibodies with a transfer efficiency of 105% (vs. 80%, P< 0.001) and 110% (vs. 90%, P< 0.001), respectively. There was a significant linear decline in maternal and cord blood RBD-specific IgG and neutralizing antibody titers as time from vaccination to delivery increased. Those who receive the mRNA COVID-19 vaccine mount an immune response that is equivalent to-if not greater than-those naturally infected by SARS-CoV-2 during pregnancy.

Immunogenicity evaluation of a novel virus-like particle vaccine candidate against SARS-CoV-2 in BALB/c.

The coronavirus disease (COVID-19) pandemic has imposed deployment of an effective vaccine as a worldwide health priority. The new variants of SARS-CoV-2 have also brought serious concerns due to virus eradiation hesitancy. In this study, we evaluated the protective immune system activity of a recombinant viral vector-based vaccine candidate encoding a fusion spike, membrane and nucleocapsid proteins, Spike (528-1273aa)-M-N, in BALB/c via two different routes of delivery, intranasal and subcutaneous. The immune responses were then assessed through specific SARS-CoV-2 antibodies, interleukin and granzyme B secretion. The outcomes showed that the IgG titer and IgA secretion was higher in intranasal route in comparison with the subcutaneous, and what is more, a higher titer of IL-4 was detected through the intranasal route, whereas IFN-γ was highly induced via the subcutaneous route. The cytotoxic cell activities were mostly achieved via subcutaneous route immunization. Vaccination with the target antigen is immunogenic and led to induction of specific antibodies. Both humoral and cellular immunity arms were well activated in immunized mice, especially through intranasal route with detectable IgA and IgG. Therefore, implication of the platform as a potential vaccine candidate has potential as a future prophylactic vaccine that guarantees further investigations for the assessment of its immunogenicity in humans.

POINT PREVALENCE OF PAEDIATRIC DENGUE VIRAL INFECTION

Objective: The study's objective is to evaluate the prevalence of dengue in the paediatric age group based on demographic data and clinical symptoms.&#x0D; Methods: The study was conducted in the Department of Paediatrics and Central Research Laboratory in a Tertiary Care Hospital in Visakhapatnam. Six months of data were collected from Jan 2022 to June 2022 and a maximum of 100 OPD/IPD cases have been registered during this study period. The entire medical history of all dengue cases, including signs and symptoms, method of diagnosis, management, length of stay, and clinical outcomes, were collected from patient information. NS1 antigen detection was performed by rapid test and Dengue ELISA was performed as a confirmatory test for the detection of IgG and IgM antibodies.&#x0D; Results: Among the 100 suspected cases of dengue reported to OPD/IPD, 83(83%) were confirmed as dengue positive, and 17(17%) were found to be non-dengue cases with ELISA. More cases were noticed in female children 52 (55%). Prevalence was more in the age group of 5-11 y 43 (52%). The majority number of cases were reported with fever 82 (98.8%) followed by symptoms like vomiting 41(49%), cold 19(16%), and cough 18(15%).&#x0D; Conclusion: Understanding dengue fever in children, including clinical symptoms, etiology, diagnostic testing, therapy, and prevention, is critical for lowering disease fatality rates.

Monkeypox infection elicits strong antibody and B cell response against A35R and H3L antigens

Monkeypox virus (MPXV) resides in two forms; mature and enveloped, and depending on it, distinct proteins are displayed on the viral surface. Here, we expressed two MPXV antigens from the mature, and one from the enveloped form, and tested their reactivity to sera of 11 MPXV recoverees while comparing to sera from recently and past vaccinated individuals. 8 out of 11 recoverees exhibited detectable neutralization levels against Vaccinia Lister. Sera from all recoverees bound strongly to A35R and H3L antigens. Moreover, the responses to A35R were significantly higher within the recoverees compared to both recently and past vaccinated donors. Lastly, A35R- and H3L-specific IgG+ B cells ranging from 0.03-0.46% and 0.11-0.36%, respectively, were detected in all recoverees (A35R), and in 9 out of 11 recoverees (H3L). Therefore, A35R and H3L represent MPXV immune targets and could be used in a heat-inactivated serological ELISA for the identification of recent MPXV infection.

Detection of IgM, IgG, IgA and neutralizing antibody responses to SARS-CoV-2 infection and mRNA vaccination.

Introduction. One correlate of immunity for coronavirus disease 2019 (COVID-19) is the laboratory detection of anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies. These tests are widely implemented for clinical, public health, or research uses.Hypothesis/Gap Statement. Antibody responses by all classes of immunoglobulins may form from infection and vaccination, but few studies have performed direct head-to-head comparisons between these groups.Aim. The objective of this study was to evaluate the serological responses in natural SARS-CoV-2 infection and mRNA-based vaccination across multiple immunoglobulin classes and a surrogate neutralizing antibody (NAb) assay.Methodology. A suite of enzyme-linked immunosorbent assays (ELISAs) was used to qualitatively assess IgA, IgM and IgG positivity and neutralizing per cent signal inhibition of sera from RT-PCR-confirmed SARS-CoV-2-infected patients, COVID-19-immunized individuals ≥2 weeks after a second dose of mRNA vaccine and a set of pre-pandemic negative samples.Results. For confirmed SARS-CoV-2 infections, seroconversion of IgA, IgM, IgG and NAb increased by week after symptom onset, with positivity reaching 100 % after the third week for every immunoglobulin class. Vaccinated individuals demonstrated 100 % IgG positivity and high per cent signal inhibition by NAb, comparable to natural infection. High specificity, ranging from 96.7-98.9 %, was observed for each assay from a set of pre-pandemic COVID-19-negative samples.Conclusion. We make use of a comprehensive and readily adoptable suite of serological assays to provide data on the humoral immune response to SARS-CoV-2 infection and vaccination. We found that infection and vaccination both elicit robust IgG, IgM, IgA and neutralizing antibody responses.

A Fiber Bragg Grating Sensor Based on Cladding Mode Resonance for Label-Free Biosensing

A fiber-optic biosensing platform based on ultra-narrowband cladding mode resonances was developed on a high-reflectivity fiber Bragg grating (FBG) for targeting biomolecular detection. The multiple cladding modes with a high sensitivity to the refractive index (RI) were excited in the FBG by coupling between the forward-propagating guided core mode of the multimode fiber and the backward-propagating guided cladding mode of the FBG without any damage to the fiber structure or any change to the standard FBG manufacturing process. The full width at half maximum and the Q-factor of the typical cladding mode resonance operation of the proposed sensor are 80 pm and 19,270, respectively, which are better than those of most fiber-optic biosensors reported to date. In addition, the FBG sensor demonstrated a high sensitivity in protein detection and a high selectivity in serum sample assays. The sensitivity of this sensor was further increased simply by coating it with graphene oxide (GO) sheets on the sensing surface without using a signal amplification strategy. Furthermore, an ultra-low limit of detection (LOD) of 32 pM was obtained by the GO-coated FBG sensor for IgG detection. The proposed FBG sensor provides a competitive fiber-optic platform for biomolecular detection. It has a great potential for applications in label-free biosensing.

Monitoring SARS-CoV-2 seroprevalence over time among pregnant women admitted to delivery units: Suitability for surveillance.

To determine SARS-CoV-2 seroprevalence over time and risk factors among pregnant women at delivery in São Paulo, Brazil; and to evaluate the suitability of pregnant women as a sentinel population for SARS-CoV-2 serosurveillance. Unselected consecutive pregnant women presenting at the labor ward of a single large hospital between July 20th 2020 to February 21st 2021 were enrolled and tested for SARS-CoV-2 serology using two assays: the rapid chromatic Wondfo One Step (for total IgA and IgG detection) and Roche Elecsys assay (detecting anti-nucleoprotein [N] IgG). SARS-CoV-2 seroprevalence was computed as smooth spline function over time with 95% confidence intervals (CI). Risk factors were evaluated for positivity by each assay. We compared timepoint seroprevalence by the two assays with four concomitant community household surveys (HHS), in which the Roche assay was used, to determine the sensitivity and relevance of the pregnant women population as sentinel population. Overall SARS-CoV-2 seroprevalence was 28.9% (221/763) by Roche and 17.9% (137/763) by Wondfo. Reported symptoms experienced during pregnancy were all significantly correlated with being SARS-CoV-2 seropositive at delivery with any assay (with odds-ratios ranging from 3.0 [95% CI: 2.1-4.3] for coryza to 22.8 [95% CI: 12.3-46.6] for ageusia). Seropositivity by either assay was high in women at delivery in the early period of the pandemic (June 2020), compared with seropositivity in women from the concomitant HHS: 44.1% (95% CI: 21.8-66.4) for Roche, 54.1% (30.9-78.5) for Wondfo, versus 11.4% (95% CI: 9.2-13.6) for HHS. For later periods (October 2020 and January 2021), the seropositivity in women at delivery measured by Roche corresponded well with the prevalence found among women in the HHS using the same assay, whilst prevalence measured by Wondfo dropped. Women at delivery represent a highly exposed and readily accessible population for sentinel surveillance of emerging infections such as SARS-CoV-2.

Клинико-лабораторные, инструментальные, морфологические особенности гангренозного и флегмонозного аппендицита у пациентов с COVID-19-инфекцией

Objective. To present clinical, laboratory, instrumental and morphological features of phlegmonous and gangrenous appendicitis in patients with COVID-19 infection. Patients and methods. During the period from March 2020 to August 2022, 23 patients were observed with positive results of testing of nasopharyngeal sample for SARS-CoV-2 RNA and performed appendectomy. The age group of children is 8–17 years old (the average age is 14.1 years), there were 13 boys, 10 girls. The premorbid state is not burdened. The rationale for appendectomy was: acute abdominal pain, local soreness on the right side of the abdomen, their preservation until the surgical removal of the appendix and their disappearance after surgery. During ultrasound of the abdominal cavity, all patients had ECHO signs of inflammatory transformation of the vermiform process. The disease was accompanied by repeated vomiting on the background of subfebrile (13 children) or normal temperature (10 children). Respiratory syndrome is poor, 4 children had abdominal pain, 9 children had hyperemia of the pharynx, chest CT did not reveal inflammatory changes. Repeated virological examination showed the positive result in 3 of 9 examined; EIA for IgM and IgG detection was positive in 7 (out of 7 examined) with diagnostic titers in 3 patients. During histological examination, the length of the removed appendixes is ~8.1 cm (4–12.5 cm), partial or complete desquamation of the epithelium, massive infiltration by lymphocytes or neutrophils, circulatory disorders, in some cases (gangrenous appendicitis) – the phenomena of thrombosis of small vessels, mesenteriolitis, periappendicitis. Typical E. Coli was found in 4 out of 23 abdominal fluid samples. Conclusion. The complex of clinical, laboratory, instrumental and morphological data confirmed the diagnosis of appendicitis and the course of COVID-19, indicating the need for surgical intervention and the appointment of antiviral therapy, noting the need to continue the study. Key words: phlegmonous, gangrenous appendicitis, children, laparoscopic removal, ultrasound of abdomen, morphological examination of the appendix

Evaluation of two SARS-CoV-2 IgG/IgM Rapid Tests in Capillary Blood Samples

As COVID-19 has spread rapidly around the world, it has become essential to detect the virus quickly and accurately for disease prevention and control. Therefore, in response to the COVID-19 pandemic, the need for rapid serological point-of-care test has increased. Recently, many antibody tests have been developed to detect IgM and/or IgG to SARS-CoV-2 in human blood. The authors conducted a prospective study to evaluate the performance of a rapid chromatographic immunoassay and a fluorescent immunoassay for the qualitative detection of specific antibodies, IgM and IgG to SARS-CoV-2 in capillary blood samples, compared to the real-time RT-PCR. The subjects included 70 patients who were confirmed positive by real-time RT-PCR and 70 people who were negative. STANDARD Q COVID-19 IgM/IgG Plus Test (chromatographic immunoassay) and Fluorescent immunoassay for IgM and IgG to SARS-CoV-2 (fluorescent immunoassay) were performed using capillary blood samples. Based on the results of real-time RT-PCR assay, clinical sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of two rapid tests were investigated. And the agreement rate between two rapid tests was also presented. Sensitivity, specificity, PPV and NPV of the chromatographic immunoassay were 82.9%, 98.6%, 98.3%, and 85.2%, respectively. At more than 7 days after the onset of symptoms, sensitivity increased to 87.3%. Sensitivity, specificity, PPV, and NPV were 81.4%, 100.0%, 100.0%, and 84.3%, respectively, for the fluorescent immunoassay. At more than 7 days after the onset of symptoms, sensitivity increased to 85.7%. The agreement rate of the two tests was 97.1%. STANDARD Q COVID-19 IgM/IgG Plus Test and STANDARD F COVID-19 IgM/IgG Combo FIA turned out very specific and sensitive enough to detect individuals infected to SARS-CoV-2. Also, these tests were simple, fast, visually interpretable, and required a small amount of capillary whole blood.

Biosensors based on DNA-functionalized CdTe quantum dots for the enhanced electrochemical detection of human-IgG.

Electrochemical detection of human-IgG via biosensors is vital in clinical diagnostics, owing to their simple equipment, facile operation, high selectivity, economical, short diagnostic time, fast response, and easy miniaturization, but the need to improve sensitivity for protein detection is still a barrier limiting its wider practical applications. A hypersensitized electrochemical biosensor based on steric effects for IgG detection was developed in this work. The results indicate that IgG-modified sig-DNA attached to CdTe quantum dots (CdTe-sig-DNA) limited the ability of CdTe-sig-DNA or CdTe-sig-DNA-IgG conjugate to hybridize through the captured DNA strand (cap-DNA) immobilized on a chitosan/nitrogen-doped carbon nanocomposite (CS/N-C) modified glassy carbon electrode surface (GCE). The concentration of IgG based on CdTe concentration was detected by differential pulse anode stripping voltammetry (DPASV) on the electrode surface. The efficiency for hybridizing CdTe-sig-DNA with cap-DNA was found to be logarithmically inverse to the concentration of IgG attached. A highly sensitive and selective detection of IgG from 5 pM to 50 μM with a relatively low detection limit of 1.7 pM was achieved. Therefore, the steric hindrance effect of IgG limited the quantity of DNA that could be functionalized on CdTe QDs, significantly improving the signal, and providing a practical strategy for the clinical analysis of IgG.

Ultra-stable CsPbBr3@PbBrOH nanorods for fluorescence labeling application based on methylimidazole-assisted synthesis.

The extension application of perovskites in aqueous media such as bioassays requires the development of a water-stable perovskite with a simple preparation process and low cost. However, the degradation of perovskites in aqueous solution is still a thorny problem. Here, we develop a methylimidazole-assisted two-step synthesis protocol to prepare CsPbBr3@PbBrOH nanorods with superior water stability and remarkable optical properties at room temperature. The synergy of 2-methylimidazole (2-MIM), an N-donor ligand, with water can not only facilitate CsPbBr3 formation and suppress CsPb2Br5 or Cs4PbBr6 formation, but also promote the formation of a PbBrOH shell capping CsPbBr3. 2-MIM is ionized into 2-MIM- in DMF and 2-MIM+ in water. They passivated the surface defects and changed the crystallization environment, leading to water-stable CsPbBr3@PbBrOH. The obtained CsPbBr3@PbBrOH nanorods can still maintain 91% PL intensity after being stored in water for more than 2 months. Furthermore, the CsPbBr3@PbBrOH nanorods show excellent stability in polar solvents, water, and phosphate buffer solution in a wide pH range, as well as better thermal and irradiation stability. In addition, the CsPbBr3@PbBrOH nanorods are further functionalized with polydopamine (PDA) for biomolecular immobilization and immunoassay studies. The resulting assay shows a detection limit of 0.003 ng mL-1 for IgG detection, illustrating important progress towards expanding fluorescence labeling application of perovskite nanomaterials for immunoassays.

D614G SARS-CoV-2 Pseudovirus Infectivity and Binding of Spike Protein to the ACE2 Receptor Inversely Correlates with Serum SARS-CoV-2-Specific IgG Levels.

Understanding the functional characteristics of antibodies produced against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) will assist in the determination of disease outcomes for this virus. In this study, the ability of antibodies to inhibit viral entry into the host cell through the interaction of the receptor binding domain of the viral spike protein and the angiotensin-converting enzyme 2 receptor on the human cell surface was investigated. The SARS-CoV-2 IgG levels in 20 SARS-CoV-2 positive patients were measured using an enzyme-linked immunosorbent assay, and the samples were further analyzed using a functional binding assay. Inhibition of viral infectivity was also measured using a pseudovirus neutralization assay against a D614G SARS-CoV-2 virus strain. A significant correlation between IgG levels and neutralizing antibody 50% inhibitory concentration (IC50) titers was observed (p < 0.05). Similarly, the IC50 titers obtained in the neutralization and binding assays were significantly correlated (p < 0.001). Varying levels of IgG and IC50 titers were observed for the SARS-CoV-2 antibody-positive samples, with one sample not showing any neutralizing capability despite detectable IgG levels. Gender comparisons showed no statistical differences in any of the assays. These results suggest that increased SARS-CoV-2 IgG levels correlate with greater protection against the entry of the virus into cells; however, further investigations in larger studies are needed to confirm the correlates of protection.

Evaluation of Two Adjuvant Formulations for an Inactivated Yellow Fever 17DD Vaccine Candidate in Mice.

The attenuated yellow fever (YF) vaccine is one of the most successful vaccines ever developed. After a single dose administration YF vaccine can induce balanced Th1/Th2 immune responses and long-lasting neutralizing antibodies. These attributes endorsed it as a model of how to properly stimulate the innate response to target protective immune responses. Despite their longstanding success, attenuated YF vaccines can cause rare fatal adverse events and are contraindicated for persons with immunosuppression, egg allergy and age < 6 months and >60 years. These drawbacks have encouraged the development of a non-live vaccine. The aim of the present study is to characterize and compare the immunological profile of two adjuvant formulations of an inactivated YF 17DD vaccine candidate. Inactivated YF vaccine formulations based on alum (Al(OH)3) or squalene (AddaVax®) were investigated by immunization of C57BL/6 mice in 3-dose or 2-dose schedules, respectively, and compared with a single dose of attenuated YF virus 17DD. Sera were analyzed by ELISA and Plaque Reduction Neutralization Test (PRNT) for detection of total IgG and neutralizing antibodies against YF virus. In addition, splenocytes were collected to evaluate cellular responses by ELISpot. Both inactivated formulations were able to induce high titers of IgG against YF, although neutralizing antibodies levels were borderline on pre-challenge samples. Analysis of IgG subtypes revealed a predominance of IgG2a associated with improved neutralizing capacity in animals immunized with the attenuated YF vaccine, and a predominance of IgG1 in groups immunized with experimental non-live formulations (alum and AddaVax®). After intracerebral (IC) challenge, attenuated and inactivated vaccine formulations showed an increase in neutralizing antibodies. The AddaVax®-based inactivated vaccine and the attenuated vaccine achieved 100% protection, and alum-based equivalent formulation achieved 70% protection.

Diagnostic performance of the rapid test for the detection of NS1 antigen and IgM and IgG anti-antibodies against dengue virus.

Motivation for the study: search for accessible and efficient new diagnostic alternatives for the detection of the disease caused by the dengue virus. Main findings: good efficiency of the rapid test during the first days of the disease. As well as its high power to discriminate against other similar mosquito-borne diseases such as Zika and Oropuche. Implications: it could be applied as a screening test in endemic regions that do not have equipment or trained personnel to perform sophisticated and/or complex diagnostic tests. Strengthening public health policies in epidemiological surveillance, early diagnosis and timely treatment. To assess the diagnostic performance of the SD dengue DUO rapid test (Inyecta) for the detection of NS1, IgM and IgG in comparison to the ELISA test. . This is a diagnostic test evaluation that included 286 serum samples from patients with symptomatology attributable to dengue from endemic areas of Peru. The samples were analyzed by ELISA and the SD dengue DUO rapid test (Inyecta) for IgM, NS1 and IgG at the Instituto de Investigación Nutricional in Lima. . The sensitivity of the rapid test was 68.0% for NS1 and IgM, and 86.0% for IgG, improving to 75.0% and 81.0% for NS1 and IgM, respectively, during the first three days. The specificity for all three analytes was greater than 87.0%. The concordance of the results, measured by the Kappa coefficient for the three analytes, was good and no cross-reaction with other arboviruses was found. . The SD dengue DUO rapid test allows detection of NS1, IgM and IgG with adequate sensitivity and specificity. Sensitivity for IgM and NS1 increases when detected during the first three days of symptoms. Therefore, we recommend its implementation in primary care centers for early and timely diagnosis.

Longitudinal characterisation of B and T-cell immune responses after the booster dose of COVID-19 mRNA-vaccine in people with multiple sclerosis using different disease-modifying therapies

BackgroundThe decline of humoral response to COVID-19 vaccine led to authorise a booster dose. Here, we characterised the kinetics of B-cell and T-cell immune responses in patients with multiple sclerosis...

Enhanced Serum IgG Detection Potential Using 38KD-MPT32-MPT64, CFP10-Mtb81-EspC Fusion Protein and Lipoarabinomannan (LAM) for Human Tuberculosis.

For the rapid, reliable, and cost-effective methods of tuberculosis (TB) auxiliary diagnosis, antibody (Ab) detection to multiple antigens of Mycobacterium tuberculosis (Mtb) has great potential; however, this methodology requires optimization. We constructed 38KD-MPT32-MPT64, CFP10-Mtb81-EspC, and Ag85B-HBHA fusion proteins and evaluated the serum Ab response to these fusion proteins and to lipoarabinomannan (LAM) by ELISA in 50 TB patients and 17 non-TB subjects. IgG responses to the three fusion proteins and to LAM were significantly higher in TB patients, especially in Xpert Mtb-positive TB patients (TB-Xpert+), than in non-TB subjects. Only the anti-38KD-MPT32-MPT64 Ab showed higher levels in the Xpert Mtb-negative TB patients (TB-Xpert-) than in the non-TB, and only the anti-LAM Ab showed higher levels in the TB-Xpert+ group than in the TB-Xpert- group. Anti-Ag85B-HBHA Ab-positive samples could be accurately identified using 38KD-MPT32-MPT64. The combination of 38KD-MPT32-MPT64, CFP10-Mtb81-EspC, and LAM conferred definite complementarity for the serum IgG detection of TB, with relatively high sensitivity (74.0%) and specificity (88.2%). These data suggest that the combination of 38KD-MPT32-MPT64, CFP10-Mtb81-EspC, and LAM antigens provided a basis for IgG detection and for evaluation of the humoral immune response in patients with TB.

Application of Toxoplasma gondii-specific SAG1, GRA7 and BAG1 proteins in serodiagnosis of animal toxoplasmosis.

Toxoplasmosis is a zoonotic disease caused by the obligate intracellular protozoan parasite T. gondii which is widely prevalent in humans and animals worldwide. The diagnosis of toxoplasmosis and distinguishing acute or chronic T. gondii infections have utmost importance for humans and animals. The TgSAG1, TgGRA7, and TgBAG1 proteins were used in the present study to develop the serological rSAG1-ELISA, rGRA7-ELISA and rBAG1-ELISA methods for the testing of T. gondii specific IgG and IgM antibodies and differentiating acute or chronic toxoplasmosis in 3733 animals, including Tibetan sheep, yaks, pigs, cows, cattle, horses, chickens, camels and donkeys from the Qinghai-Tibetan Plateau. The ELISA tests showed that the overall positivity of IgG antibody was 21.1% (786/3733), 15.3% (570/3733) and 18.2% (680/3733) for rSAG1-, rGRA7- and rBAG1-ELISA, respectively, and the positivity of IgM antibody was 11.8% (439/3733), 13.0% (486/3733) and 11.8% (442/3733) for rSAG1-, rGRA7- and rBAG1-ELISA, respectively. A total of 241 animals (6.5%) positive for all rSAG1-, rGRA7- and rBAG1-IgG were found in this study, and the 141 animals (3.8%) tested were anti-T. gondii IgM positive in all three ELISAs. Moreover, the 338, 284 and 377 animals were IgG positive in rSAG1 + rGRA7-, rBAG1 + rGRA7- and rSAG1 + rBAG1- ELISAs respectively, and the 346, 178 and 166 animals in rSAG1 + rGRA7-, rBAG1 + rGRA7- and rSAG1 + rBAG1-ELISAs were IgM positive respectively. The results confirmed that the application of SAG1, GRA7, and BAG1 recombinant antigens could successfully be used in the detection of specific IgG and IgM antibodies for distinguishing between acute or chronic T. gondii infections. It is inferred that the forms in which current animal species in the plateau area were infected with T. gondii, and the period of infection or the clinical manifestations of the current infections may be different. The present study provides substantial clinical evidence for the differential diagnosis of toxoplasmosis, and the classification of acute and chronic T. gondii infections.