Antibody ELISA Research Articles

Co-infection of Hepatitis C among HIV-infected patients: A cross-sectional study from A University Teaching hospital in Anambra State, Nigeria

Hepatitis C virus co-infection (HCV) with HIV is regarded as a significant public health risk despite being less frequent than hepatitis B co-infection with HIV. This observation results from HIV's impact on the HCV life cycle and, ultimately, the hepatic system. In this study, potential HCV co-infection was examined in connection to the socio-demographic characteristics of HIV-positive patients attending an HIV clinic at a University Teaching Hospital in Awka, Anambra State, Nigeria. Between September 2021 and June 2022, 255 HIV patients who consented to participate in the study had their HCV levels checked using the anti-HCV antibody ELISA kit (DIA.PRO, Italy). Using the Partec CyFlow, CD4 counts were calculated. During routine investigations, blood samples (approximately 5ml) were aseptically collected into sterile EDTA vials, and plasma samples were obtained by centrifugation. With the aid of Abbott Real-Time HIV-1 Assay US methodology, plasma viral loads (PVL) were also calculated. A total of 11 (4.3%) HIV-positive people were also confirmed to have HCV infection, with women making up the majority (4.8%). Most of those affected were between the ages of 31 and 40 (6.8%). With marital status, HCV co-infection was found more amongst married patients, with a rate of 4.8%. The prevalence of HIV/HCV co-infection was higher (4.8%) in those with CD4 cell counts under >350 cells per ml than others. Most patients (3.1%) did not have their viral RNA detected (TND). Self-employed infected persons and those with secondary school educational backgrounds had the highest HIV/HCV prevalence rate of 6.6% and 7.6%, respectively. The study's findings show no statistically significant relationships between the patients' socio-demographic traits and HCV. Also, HCV co-infection prevalence was significantly low in the study participants.

Goats with low levels of AAV antibody may serve as candidates for large animal gene therapy

AAV vector-mediated gene therapy has been proposed as a feasible strategy for several eye diseases. However, AAV antibodies in the serum prior to treatment hinder the transduction efficiency and thus the therapeutic effect. Therefore, it is necessary to evaluate AAV antibodies in the serum before gene therapy. As large animals, goats are more closely related to humans than rodents and more economically available than nonhuman primates. Here, we first evaluated the AAV2 antibody serum level in rhesus monkeys before AAV injection. Then, we optimized a cell-based neutralizing antibody assay for detecting AAV antibodies in the serum of Saanen goats and evaluated the consistency of the cell-based neutralizing antibody assay and ELISA for goat serum antibody evaluation.The cell-based neutralizing antibody assay showed that the percentage of macaques with low antibody levels was 42.86%; however, there were no macaques with low antibody levels when the serum was evaluated by ELISA. The proportion of goats with low antibody levels was 56.67% according to the neutralizing antibody assay and 33. 33% according to the ELISA, and McNemar's test showed that the results of the two assays were not significantly different (P = 0.754), but that their consistency is poor (Kappa = 0.286, P = 0.114). Moreover, longitudinal evaluation of serum antibodies before and after intravitreal injection of AAV2 in goats revealed that the level of AAV antibodies increased and transduction inhibition subsequently increased, as reported in humans, indicating that transduction inhibition should be taken into account at different stages of gene therapy. In summary, starting with an evaluation of monkey serum antibodies, we optimized a detection method of goat serum antibodies, providing an alternative large animal model for gene therapy, and our serum antibody measurement method may be applied to other large animals.

COMPARISON OF ANTIBODY DETECTION BY RAPID DIAGNOSTIC KIT AND ELISA TEST FOR DIAGNOSIS OF HEPATITIS C VIRUS INFECTION

Background: Hepatitis C virus (HCV) infection is a major cause of healthcare problem along with burden of health management care cost of international concern. Hepatitis C is a serious threat with long term complication. The present study is undertaken to compare rapid card test based on Immunochromatography principle with ELISA test for detection of anti HCV antibodies. In the absence of vaccine, primary prevention of Hepatitis C should be targeted to reduce transmission of virus. High risk people should be provided with education, counselling and screening. A cross-sectional study was done at Dept. of Microbiology, M.G.M.M.C, Indore Method: for one year comprising of 500 samples after getting Institutional Scientic Research and Ethics Committee approval. Rapid card and ELISA tests were performed in the samples received for detection of Anti-HCV ab. Out of 500 blood samples test Result: ed on Rapid card 22 were positive while 478 were negative. Further testing on same 500 samples with ELISA test 22 were positive and 478 were negative. Discussion: Present study ndings suggest that for screening of anti HCV antibody ELISA and Rapid card tests both can be used as diagnostic tool depending upon facilities available in the laboratory. Rapid card Conclusion: tests were found to be equally diagnostic as compared to ELISA test and hence can be recommended in screening of HCV

Obtaining and Characterization of Hybridomas Producing Monoclonal Antibodies against Coronavirus SARS-CоV-2

The aim of the work was to obtain and characterize hybridomas producing monoclonal antibodies to antigens of coronavirus SARS‑CoV‑2, promising for the construction of diagnostic immunochemical tests. Materials and methods. Recombinant nucleocapsid and receptor binding fragment of spike protein of SARS‑CoV‑2 were used for immunization of BALB/c mice. Antigens were absorbed on aluminium hydroxide gel and injected subcutaneously to BALB/c mice at a 7-day-interval. Immune splenocytes and myeloma cells SP2/0-Ag14 were fused by polyethylene glycol 1450. Cell cultures producing specific antibodies against nucleocapsid and receptor binding fragment were selected applying indirect ELISA in 96-well plates sensitized by desired antigens. Clones of hybridomas were obtained using the method of limiting dilutions. Production properties were studied through in vitro cultivation in 24-well culture plates. Immune-ascitic fluids were collected during the cultivation of hybrid cells in peritoneal cavities of BALB/c mice. Monoclonal antibodies were purified by affinity chromatography on protein A sepharose sorbent, conjugated with horseradish peroxidase, and tested for the possibility to be used in sandwich ELISA for detection of inactivated SARS‑CoV‑2 coronavirus strain “Isolate B”. Results and discussion. As a result of hybridization and selection of clones, hybridomas producing monoclonal antibodies to nucleocapsid and receptor binding fragment of SARS‑CoV‑2 have been obtained. During the in vitro and in vivo cultivation the clones maintained the consistent proliferative and antibody producing activity. The application of monoclonal antibody 415D12 as a capture one and 411D12 antibody conjugated with horseradish peroxidase as a detector antibody in ELISA allows for identifying SARS‑CoV‑2 coronavirus at a minimum concentration of 1·103 PFU per ml.

Indirect IgG ELISA and serotype-specific neutralizing antibody titers are associated with protection against dengue in children in Cebu, Philippines

Abstract Standard measures of dengue virus (DENV) antibodies have not been established as protective against dengue. We assess whether an indirect dengue IgG ELISA (PanBio; Brisbane, QLD, Australia) and serotype-specific neutralizing antibody titers measured by a two-dilution focus reduction neutralization test (FRNT) are associated with protection against dengue. Healthy children (n = 1,214) aged 9–14 years in Cebu, Philippines were enrolled in a prospective population-based cohort study in May – June 2017 and were tested for dengue when they reported a febrile illness through March 2020. Serum samples were collected at enrollment, and all were tested by ELISA. FRNTs were performed using the WHO DENV1–4 reference strains on a random subset to quantify serotype-specific neutralizing antibodies (nAbs). Inverse probability weighting was used to adjust for subset size. The odds of dengue as predicted by antibody measures were assessed by logistic regressions and adjusted for age, gender, and residential site. Dengue naïve children were used as the reference group in all models. The probability of dengue was 9.2% (95% CI: 4.5 – 17.9) for the naïve group, 9.5% (4.8 – 17.8) for those with ELISA < 2, but 4.5% (2.8 – 7.2) for those with ELISA >= 2 (p=0.04). The probability was even lower at 1.4% (0.6 – 3.1) for those with the highest DENV1–4 geometric mean titers (GMT, p = 0.0001). Pre-vaccine nAbs to DENV2 and DENV3 were protective against a homotypic dengue case (DENV2: OR 0.28, 95% CI 0.12 to 0.65; DENV3: OR 0.27, 95% CI 0.13 to 0.57), but not heterotypic dengue. We show a commercial ELISA and serotype-specific nAbs are associated with the probability of dengue, providing useful assays for use in population and vaccine trials. This work was supported by the Philippine Department of Health, the Intramural Research Program of the National Institute of Allergy and Infectious Diseases (NIAID), and NIH Extramural Grant: P01AI106695.

Clinical validation of novel dried blood spot based collecting device using serum separation for measuring SARS-CoV-2 antibodies.

Accurate surveillance of coronavirus disease 2019 (COVID-19) incidence includes large-scale antibody testing of the population. Current testing methods require collection of venous blood samples by a healthcare worker, or dried blood spot (DBS) collection using finger prick, however this might have some logistic and processing limitations. We investigated the performance of the Ser-Col device for detecting severe acute respiratory syndrome coronavirus-2(SARS-CoV-2) antibodies using a finger prick: DBS-likecollection system that includes a lateral flow paper for serum separation and allows for automated large scale analysis.For this prospective study, adult patients with moderate to severe COVID-19 were included 6 weeks post-symptom onset. Healthy, adult volunteers were included as a negative control group. Venous blood and capillary blood using the Ser-Col device were collected and the Wantai SARS-CoV-2 total antibody ELISA was performed on all samples.We included 50 subjects in the study population and 49 in the control group. Results obtained with venous blood versus Ser-Col capillary blood showed 100% sensitivity (95% CI: 0.93-1.00) and 100% specificity (95% CI: 0.93-1.00).Our study shows the feasibility of SARS-CoV-2 total antibody screening using a standardized DBStechnique with semiautomated processing for large scale analysis.

SARS-CoV-2 Ancestral and Variant of Concern Breakthrough infections boost both cross-reactive cellular and humoral immune responses in mRNA-vaccinated Hamsters.

Abstract When mRNA vaccines were rolled out under EUA, it was observed that a single vaccination, already protected vaccinees from severe disease, even before neutralizing antibodies appearded. To better understand the underlying mechanism of protection, we vaccinated hamsters once with a suboptimal dose of Pfizer/BioNTech COVID-19 mRNA vaccine (5mg/animal). Four weeks post vaccination, animals were challenged with different variants of SARS-CoV-2. Vaccination resulted in antibody ELISA titers against SARS-CoV-2 spike protein, but antibody titers were too low for efficient neutralization of antigenically matching USA-WA1/2020 (WA1) or drifted variants of concern (Alpha, Beta, Delta and Mu). Nevertheless, suboptimal vaccination resulted in reduced morbidity for WA1 and Alpha-challenged animals and complete control of lung virus titers for WA1, Alpha and Delta but with breakthrough infection for the antigenically more distant Beta and Mu. Moreover, T cell responses were higher in vaccinated animals compared to unvaccinated ones at 5 days post infection, suggesting a recall of vaccine-primed T cells. Infection with different SARS-CoV-2 variants also back-boosted neutralizing antibody responses against the challenge virus and in the case of the variants of concern, against the antigenically distant but vaccine-matched WA1. Transcriptomic analysis of host immune responses to infection reflects both vaccination status and disease course, is further compared with lung pathology and suggests a role for interstitial lung macrophages in vaccine-mediated protection. In summary, protection after suboptimal vaccination against SARS-CoV-2 challenge in hamsters correlates with recall of both vaccine-induced B- and T-cell responses. This study was also partly funded by CRIPT (Center for Research on Influenza Pathogenesis and Transmission), a NIH NIAID funded Center of Excellence for Influenza Research and Response (CEIRR, contract number 75N93021C00014) to A.G.S and M.S. and by R01 AI160706 to M.S.

PCR and peptide based PCMV detection in pig - development and application of a combined testing procedure differentiating newly from latent infected pigs.

Porcine cytomegalovirus (PCMV) is widely distributed in pigs and difficult to detect due to latency. PCMV infection of source pigs was associated with early graft failure after cardiac and renal xenotransplantation into nonhuman primates. Importantly, PCMV infection of the first genetically modified pig heart into a human may have contributed to the reduced survival of the patient. Sensitive and reliable assays for detection of latent PCMV infection are thus indispensable. Here, we report the development of five peptide-induced rabbit antisera specific for PCMV glycoprotein B (gB) and their validation for detection of PCMV in infected pig fallopian tube (PFT) cells by immunofluorescence and electron microscopy (EM). The anti-gB antibodies were also used for detection by Western blot analysis of PCMV purified from the supernatant of infected PFT cells. Sera of infected versus non-infected pigs have been compared. In parallel, PCMV viral load in blood samples of the animals was quantified by a novel highly sensitive nested-PCR and qPCR assay. A combination of four partly overlapping peptides from the gB C-terminus was used to establish a diagnostic ELISA for PCMV gB specific pig antibodies which is able to differentiate infected from non-infected animals and to quantify maternal antibodies in neonates. The combination of a highly sensitive nested PCR for direct virus detection with a sensitive peptide-based ELISA detecting anti-PCMV gB-antibodies, supplemented by Western blot analysis and/or immunohistochemistry for virus detection will reliably differentiate pigs with active infection, latently infected pigs, and non-infected pigs. It may significantly improve the virologic safety of xenotransplantation.

An ELISA system for tetanus toxoid potency tests: An alternative to lethal challenge

For a long time, a widely used method for tetanus toxoid (Ttd) potency has been the challenge test, in which animals are immunized and then challenged with tetanus toxin in lethal or non-lethal way. In the context of animal welfare, an alternative is desired because the method causes unsustainable distress to animals.We aimed to replace the system for describing test results, in which scores are assigned to symptoms exhibited by challenged animals, with scores assigned to antibody ELISA titers in immunized mouse sera. The potency values and confidence intervals calculated by the absorbance score system were equivalent to those calculated by the symptom score system. We also attempted to utilize the raw ELISA absorbance instead of the assigned absorbance score and obtained similar results. ELISA may serve as an alternative to the lethal challenge for Ttd potency tests, not only in Japan but also in other countries in which mouse challenge tests are employed.

Pre-pandemic SARS-CoV-2-specific IFN-γ and antibody responses were low in Ugandan samples and significantly reduced in HIV-positive specimens.

We investigated whether prior SARS-CoV-2-specific IFN-γ and antibody responses in Ugandan COVID-19 pre-pandemic specimens aligned to this population's low disease severity. We used nucleoprotein (N), spike (S), NTD, RBD, envelope, membrane, SD1/2-directed IFN-γ ELISpots, and an S- and N-IgG antibody ELISA to screen for SARS-CoV-2-specific cross-reactivity. HCoV-OC43-, HCoV-229E-, and SARS-CoV-2-specific IFN-γ occurred in 23, 15, and 17 of 104 specimens, respectively. Cross-reactive IgG was more common against the nucleoprotein (7/110, 15.5%; p = 0.0016, Fishers' Exact) than the spike (3/110, 2.72%). Specimens lacking anti-HuCoV antibodies had higher rates of pre-epidemic SARS-CoV-2-specific IFN-γ cross-reactivity (p-value = 0.00001, Fishers' exact test), suggesting that exposure to additional factors not examined here might play a role. SARS-CoV-2-specific cross-reactive antibodies were significantly less common in HIV-positive specimens (p=0.017; Fishers' Exact test). Correlations between SARS-CoV-2- and HuCoV-specific IFN-γ responses were consistently weak in both HIV negative and positive specimens. These findings support the existence of pre-epidemic SARS-CoV-2-specific cellular and humoral cross-reactivity in this population. The data do not establish that these virus-specific IFN-γ and antibody responses are entirely specific to SARS-CoV-2. Inability of the antibodies to neutralise SARS-CoV-2 implies that prior exposure did not result in immunity. Correlations between SARS-CoV-2 and HuCoV-specific responses were consistently weak, suggesting that additional variables likely contributed to the pre-epidemic cross-reactivity patterns. The data suggests that surveillance efforts based on the nucleoprotein might overestimate the exposure to SARS-CoV-2 compared to inclusion of additional targets, like the spike protein. This study, while limited in scope, suggests that HIV-positive people are less likely than HIV-negative people to produce protective antibodies against SARS-CoV-2.

Epidemiological and demographic characteristics of dengue at a tertiary care center in Kutch region during the year 2019

Dengue is the most rapidly spreading mosquito borne viral disease of mankind, with a 30 fold increase in global incidence over the last five decades. It is a major public health concern throughout the tropical and subtropical regions of the world. The present study was conducted with objective to study epidemiological and demographic characteristics of dengue infection during the year 2019 in Kutch region, Gujarat, India.The study was carried out from January 2019 to December 2019 at Department of Microbiology, Gujarat Adani Institute of Medical Sciences, G.K. General Hospital, Bhuj. The patients having suspected dengue fever was based on standard criteria like presentation of febrile illness of 2-7 days duration were included in this study.A total of 1509 blood samples were collected during study period and serologically tested for dengue NS1 antigen and IgM antibody by capture ELISA testing method. Total 1509 blood samples were tested by ELISA for NS1 antigen and/or IgM antibody as per the protocols and personal, demographic and clinical details of each patient was recorded. Out of 1509 cases 761 tested positive for dengue. Amongst all positive cases 533(47%) were tested positive for NS1 antigen which helped in early diagnosis of dengue. Rise of dengue cases started after the month of October and falls down by the end of December. Analysis of this data revealed that age group 00-10 years had maximum dengue cases. Male cases 418(55%) were more than females 343(45%). Dengue cases were high in urban area.The present study reported that dengue mainly affected children, males and urban population. Perennial occurrence with seasonal increase during monsoon and post monsoon moths was reported. Effective implementation of vector control measures through efforts toward vector breeding source reduction and with the use of personal prophylactic measures against mosquito bites will help in reducing the dengue prevalence in the community.

High and Persistent Anti-GM1 Antibody Titers Are Associated With Poor Clinical Recovery in Guillain-Barré Syndrome.

Guillain-Barré syndrome (GBS) is an acute immune-mediated polyradiculoneuropathy that may follow a preceding infection inducing a cross-reactive antibody response to glycosphingolipids in peripheral nerves. The immune response in GBS is considered to be short lasting, explaining its monophasic clinical course. However, the disease course varies between patients, and residual deficits frequently occur. The duration of the antibody response has not been defined extensively in GBS, and the persistence of these antibodies may impair clinical recovery. The aim of this study was to determine the titer course of serum antibody titers to the ganglioside GM1 in relation to clinical course and outcome in patients with GBS. Acute-phase sera from patients with GBS included in previous therapeutic trials were screened for anti-GM1 IgG and IgM antibodies in ELISA. Anti-GM1 antibody titers were determined in sera collected at entry and during a 6-month follow-up. Clinical course and outcomes were compared between groups based on the titer course. Anti-GM1 antibodies were detected in 78 (20.7%) of 377 included patients. The anti-GM1 IgG and IgM antibody titer course was highly variable between patients. A subset of anti-GM1-positive patients had persistent anti-GM1 antibodies at 3 months (n = 27/43 [62.8%]) and 6 months (n = 19/41 [46.3%]). Patients with a high anti-GM1 IgG and IgM titer at entry recovered more slowly and less complete than anti-GM1-negative patients (IgG: p = 0.015, IgM: p = 0.03). High vs low IgG titers were independently associated with poor outcome after correcting for known prognostic factors (p = 0.046). Among patients with a high anti-GM1 IgG titer at entry, a slow titer decline was associated with poor outcome at 4 weeks (p = 0.003) and 6 months (p = 0.032). Persistent high IgG titers at 3 and 6 months were associated with poor outcome at 6 months (3 months: p = 0.022, 6 months: p = 0.004). High anti-GM1 IgG and IgM antibody titers at entry and persistent high anti-GM1 IgG antibody titers are associated with poor outcome in patients with GBS. Antibody persistency indicates ongoing antibody production long after the acute disease state in GBS. Further research is required to determine whether antibody persistency interferes with nerve recovery and is a target for treatments.

Knob domain of Fiber 2 protein provides full protection against fowl adenovirus serotype 4

Highly pathogenic fowl adenovirus serotype 4 (FAdV-4) is an acute infectious disease with severe economic impact, causing chicken hepatitis hydropericardium syndrome (HHS) and high mortality. In the present study, we evaluated the immunogenicity of the recombinant Fiber2-knob protein (F2-Knob) as an FAdV-4 candidate subunit vaccine in 14-day-old SPF chickens. The knob domain is the functional region of the viral surface protein Fiber2. The protein was expressed in Escherichia coli and was administered a single immunization with different vaccine doses. The protective efficacy was evaluated by mortality, clinical symptoms, virus shedding and histopathological examinations after challenged with the FAdV-4. The results showed that the level of ELISA antibodies of the chickens immunized with Fiber2-knob protein was significantly higher than that of the chickens immunized with an inactivated vaccine against FAdV-4. The antibody value of the immunized Fiber2-knob protein was positively correlated with the increase in immunization dose. The challenge experiment showed that the F2-Knob protein provided full protection against virulent FAdV-4 challenge and significantly reduced viral shedding. These results suggest that F2-Knob protein could be a novel vaccine candidate provide insights to control FAdV-4.

Difficulties of Q Fever Diagnostic Verification at Negative PCR Testing Results

Aim of the work: to demonstrate the difficulty of verifying the diagnosis of Q fever with negative results of PCR (DNA of Coxiella burnetii) in the blood and to assess the occurrence of serological markers among patients selected for this study based on a combination of clinical and epidemiological data. Materials and methods: plasma/serum samples of 111 patients according to clinical and epidemiological data studied due ELISA and PCR for specific antibodies to Coxiella burnetii and DNA of pathogen. Additionally, in the presence IgG to C. burnetii phase II, IgG / IgA to phase I and the avidity of specific IgG were studied. Results: the specific antibodies to C. burnetii antigens at negative results of PCR detected in 10 cases. The article provides the description of three clinical cases for demonstration of difficulties of coxiellosis diagnosis with analysis of serological profiles, titers and avidity of antibodies. Conclusion: the results of the study indicate that negative results of PCR testing do not exclude C. burnetii infection. For patients who, according to clinical and epidemiological data, Q fever is not excluded, it is advisable to prescribe a complex of laboratory tests to verify the diagnosis, which includes not only studies of the pathogen’s DNA, but also specific antibodies. To clarify the stage of the disease and reduce the risk of developing complications of coxiellosis, it is necessary to monitor the dynamics of antibody titers to C. burnetii in phase I and II phase states differentially.

Abstract 330: EP300 loss drives tumorigenesis in bladder cancer via activation of IL-6/JAK/STAT3 signaling

Abstract Bladder cancer (BLCA) is the sixth most common cancer in the U.S with 83,730 new cases, accounting for 17,200 deaths in the year 2021 and 212,536 deaths worldwide. EP300, a histone acetyltransferase gene, is mutated in ~17% of BLCA and in other cancer types such as endometrial cancer. The mechanisms by which EP300 mutations contribute to tumorigenicity of BLCA is not currently understood. To determine the phenotypic effects of loss-of-function EP300 mutations, we generated isogenic BLCA cells with EP300 knocked out (KO) using CRISPR/CAS9. Conspicuous changes in phenotypes consistent with more aggressive forms of cancer were observed in EP300-null BLCA clones, including luminal-basal histotype plasticity, anchorage-independent growth, enhanced in vitro and in vivo cell proliferation and enhanced invasion in Boyden chamber assays. To identify mechanisms whereby EP300 KO promotes increased oncogenicity, we performed phospho Tandem Tag Mass Spectrometry (TMT MS) using lysates from parental and EP300 KO cells. A notable finding was significantly increased expression of phosphorylated STAT3 (pSTAT3), which we subsequently confirmed by western blot. siRNA knockdown and selective inhibitor experiments indicated JAK1 as the upstream activator of pSTAT3 in BLCA cells. Studies of conditioned media (CM) from the EP300 KO cells using ELISA and neutralizing antibodies demonstrated that secreted IL-6 ligand and soluble IL-6R in the CM drove STAT3 activation through the IL-6 family common signaling subunit, gp130, i.e. trans-signaling. BLCA RT112 cells express an oncogenic FGFR3-TACC3 fusion and are sensitive to the FDA-approved FGFR kinase inhibitor erdafitinib. pSTAT3 expression was not dependent on FGFR3 signaling in EP300 KO RT112 cells, and EP300 KO was sufficient to confer erdafitinib resistance in a BLCA context. In sum, our results uncover that EP300 loss enhances IL-6/JAK/STAT3 signaling which promotes BLCA tumorigenesis and FGFR inhibitor resistance. Our findings elucidate a role for cytokine induced sterile inflammation in EP300-mediated tumorigenesis in bladder cancers and suggest that targeting the IL-6/JAK/STAT3 axis may represent a novel therapy strategy to overcome FGFR3 inhibitor resistance. Citation Format: Sizhi P. Gao, James A. Rodrigues, Amanda R. Sabel, Jiaqian Luo, Chen Ziyu, Xinran Tang, Eduardo A. Mascareno, Naryan Rustgi, Hikmat Al-Ahmadie, Kwanghee Kim, Eugene J. Pietzak, Gopakumar V. Iyer, David B. Solit. EP300 loss drives tumorigenesis in bladder cancer via activation of IL-6/JAK/STAT3 signaling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 330.

Controlling bovine leukemia virus in a large dairy herd by selective culling based on diagnostic testing

This intervention field trial aimed to reduce bovine leukemia virus (BLV) from a large dairy herd using a new diagnostic test to identify the most infectious cows for segregation or culling. Bovine leukemia virus infects over 40% of all dairy cattle in the United States. Similar prevalence has been reported in many other countries. Previously hidden economic impacts of BLV on the industry have only recently been recognized. The traditional method of controlling this disease was by culling all antibody-positive cattle. The availability of a new diagnostic test can identify the most infectious cows for removal. A 3,000-cow dairy farm conducted a 4-yr BLV disease control program using blood lymphocyte count (LC), BLV ELISA antibody test, and quantitative-PCR proviral load (PVL) to identify the most infectious animals for segregation or culling. The percentage of cows with a high LC was significantly decreased from 4.22 to 1.04% after the first year of selective culling of cows with high LC. Thereafter, BLV ELISA screening with PVL followup assays were used to target segregation and culling of cows with the highest PVL. After 3 yr, only 0.85% of the tested cows were BLV ELISA positive. The prevalence of animals with detectable PVL decreased from 284 to zero animals. When coupled with the results of an intervention trial in 3 small dairy farms, our findings from a large dairy herd demonstrate that BLV may be eradicated without the simultaneous culling of all ELISA-positive cows.

Sero-Detection of Sars-Cov-2 Antibodies and some Associated Risk Factors among Staff of Federal University Gusau, Zamfara State, Nigeria

This study was aimed at detecting the level of SARS-CoV-2 antibody response among the staff of Federal University Gusau, Zamfara State. A cross-sectional study design and clusteredstratified-random sampling strategy were used to sample staff of the University comprising academic, technical and non-academic staff. SARS CoV-2 antigen Rapid Tests were carried out using on the spot analysis of nasopharyngeal and blood samples of the participants using SARS-CoV-2 antigen RDT kits and total antibody ELISA kit. None of the samples collected was positive to the antigen RDT kit, however further confirmation with ELISA kit revealed a prevalence of 32.8%. This study showed that academic staffs are more exposed to SARS-CoV-2 viruses due to high incidence of antibodies detected among the academics; however, the difference in the prevalence between academic and non-academic staff was not statistically significant. Knowledge of SARS-CoV-2, belief, behaviour towards COVID19 protocols, contact history and flu-like symptoms were risk factors that predispose one to SARS-CoV-2 viruses.

Evaluation and application of a milk antibody ELISA for assessing the prevalence and incidence of bovine tuberculosis in dairy herds in Hubei Province, China

Bovine tuberculosis (bTB) is a chronic zoonotic disease that is endemic in China. Current in-vitro tests for bTB are mainly based on blood assays. Collection of samples results in some stress to the sampled cattle and associated economic losses for the herd owner. This study was designed to investigate the relationship between milk and serum antibody tests for bTB in dairy cows using 85 cows with milk and corresponding blood samples. Totally 4,395 milk samples were used to assesse the apparent (test) prevalence and incidence of bTB using the milk antibody ELISA. The association between levels of bTB milk antibody and milk quality was also evaluated. Milk and serum antibody tests showed a good correlation with a 87.5% (95% CI: 61.7%, 98.4) positive agreement and 98.7% (95% CI: 95.4, 99.8) negative agreement. The animal level lactoprevalence ranged from 0.3% (95% CI: 0, 1.2) to 33.3% (95% CI: 26.6, 40.6) in different farms and the incidence rate ranged from 0 head/cow-month (95% CI: 0, 0.02) to 0.04 head/cow-month (95% CI: 0.02, 0.07). Twenty percent of sampled farms met the criteria for bTB control in China. The prevalence on large-scale farms was lower (p < 0.001) than on small farms. The bTB milk antibody levels had a negative correlation with milk yield and a positive correlation with somatic cell count (SCC), milk protein percentage (MPP) and percentage of total solids (TS). According to this research, milk ELISA could be used as a supplement of blood samples to assist in the surveillance for bTB and for alerting control and eradication of bTB.

Point-of-Care Peptide Hormone Production Enabled by Cell-Free Protein Synthesis.

In resource-limited settings, it can be difficult to safely deliver sensitive biologic medicines to patients due to cold chain and infrastructure constraints. Point-of-care drug manufacturing could circumvent these challenges since medicines could be produced locally and used on-demand. Toward this vision, we combine cell-free protein synthesis (CFPS) and a 2-in-1 affinity purification and enzymatic cleavage scheme to develop a platform for point-of-care drug manufacturing. As a model, we use this platform to synthesize a panel of peptide hormones, an important class of medications that can be used to treat a wide variety of diseases including diabetes, osteoporosis, and growth disorders. With this approach, temperature-stable lyophilized CFPS reaction components can be rehydrated with DNA encoding a SUMOylated peptide hormone of interest when needed. Strep-Tactin affinity purification and on-bead SUMO protease cleavage yield peptide hormones in their native form that are recognized by ELISA antibodies and that can bind their respective receptors. With further development to ensure proper biologic activity and patient safety, we envision that this platform could be used to manufacture valuable peptide hormone drugs in a decentralized way.

Lumpy Skin Disease (LSD): Etiology, Pathogenesis, Prevention and Control

Lumpy skin disease (LSD) is one of the diseases that pose a threat to the livestock industry because it can cause large economic losses. The disease was first discovered in the country of Zambia in 1929. In Indonesia, LSD was first discovered in early 2022 in Indragiri Hulu Regency, Riau. This disease is caused by a virus belonging to the genus Capripoxvirus, subfamily chordopoxvirinae, family Poxviridae. The LSD virus has a limited host and does not infect non-ruminant hosts. A characteristic clinical symptom of LSD is the appearance of nodules on the skin. Diagnosis of this disease can be done by looking at typical clinical symptoms, laboratory tests such as virus isolation, serological tests (serum neutralization test, virus neutralization test (VNT), agar gel immune diffusion, indirect ELISA, and indirect fluorescent antibody technique (IFAT)), real time and conventional PCR, immunohistochemistry, LAMP, and IPMA. Prevention and control of LSD disease can be done in several ways such as vaccination, vector control, restriction of livestock traffic, strict quarantine and stamping out.