Enzyme-linked Immunosorbent Assay Test Research Articles

Epidemiological analysis of blood donors from college students in Hefei city.

College students represent a unique demographic group as they are adults no longer under direct parental care, yet often lack the institutional health support available to more established members of society, which can lead to their health needs being neglected, despite their substantial contributions to blood donation. The objective of this study is to shed light on the health status of college students in Hefei, with a specific focus on transfusion-transmitted diseases. Based on the detailed data analysis, the implementation of some constructive strategies will play a good warning role in improving clinical blood safety and promoting better health monitoring of this population in the future. The study aims to achieve two main objectives: On the one hand, to deepen our understanding of college students' participation in blood donation activities. On the other hand, to propose effective strategies for increasing youth involvement. Additionally, such research aims to afford recommendations for ensuring the safety of clinical blood supplies. Although the strategies and suggestions presented are speculative, they are grounded in the logical analysis of data and relevant theoretical frameworks. Therefore, these recommendations serve as a basis for potential future testing and implementation. The evaluation process for blood samples at the Hefei Blood Bank involves thorough serological testing for hepatitis B and C, HIV, and Treponema pallidum using both Enzyme-Linked Immunosorbent Assay (ELISA) and Nucleic Acid Testing (NAT). HIV reactive samples were further confirmed using Western Blot techniques at an external specialized laboratory. To ensure accuracy, each sample was tested with reagents from two different manufacturers. Moreover, any initially reactive sample was retested with the same reagent to eliminate potential false positives. In cases of conflicting results, alternative reagent kits are utilized for further confirmatory tests. Only samples that consistently show reactive results in consecutive tests are considered positive confirmation samples. Upon analysis of the data set, it was found that out of the total 663,105 donors, 177,089 (26.7%) were identified as college students, with the remaining 486,016 categorized as noncollege students. Our study demonstrated a notably lower prevalence of blood-transmitted infections among college students (0.48%) in contrast to the wider donor population (1.23%), underscoring their potential as safe blood donors. To uphold this level of safety and protect public health, it is crucial to increase disease prevention awareness and advocate for responsible donation practices within this specific demographic. Strengthening confidentiality protocols and expanding education on safe sexual behaviors in university settings are vital actions to cultivate an environment that promotes informed and secure blood donation. The overall group had prevalence rates of 0.44% for hepatitis B, 0.15% for hepatitis C, 0.02% for HIV, and 0.42% for Treponema pallidum. When focusing on the student cohort, the prevalence rates were 0.17% for hepatitis B, 0.04% for hepatitis C, 0.02% for HIV, and 0.23% for Treponema pallidum. Transmissibility through blood transfusion All donors (%) Students (%) Non-students (%) Any infection 1.23 0.48 1.51 HbsAg 0.44 0.17 0.54 Anti-HCV 0.15 0.04 0.19 HIV Ag/Ab 0.02 0.02 0.02 Anti-TP 0.42 0.23 0.49 Individuals with two or more infectious agents occur more than once in all categories. While it leads to numerical over-representation, such overlap is minimal and statistically insignificant.

Effects of venoactive drug therapy and ovarian vein interventions on vasoactive neuropeptide and cytokine levels in patients with pelvic venous disorders

ObjectiveTo study and compare the effects of venoactive drug (VAD) therapy and ovarian vein embolization or resection (OVE or OVR, accordingly) on the levels of vasoactive peptides and cytokines in patients with pelvic venous disorders (PeVD). MethodsThe study included 70 consecutive female patients with PeVD symptoms, such as chronic pelvic pain (CPP), dyspareunia, dysuria, and vulvar varicosities. Based on the results of clinical examination and duplex ultrasound (DUS) of the pelvic veins, the patients were allocated to the VAD therapy (n=38) or OVE/OVR (n=32). Additionally, the enzyme-linked immunosorbent assay (ELISA) tests were performed to determine levels of calcitonin gene-related peptide (CGRP), substance P (SP), interleukins 6 and 8 (IL-6, IL-8) and monocyte chemotactic protein-1 (MCP-1) after a 2-month course of VAD therapy and at 3 months after OVE/OVR. ResultsThe VAD therapy was associated with a significant decrease in CPP in 84% of patients with PeVD and isolated lesions of the parametrial veins (PVs) and uterine veins (UVs). VAD had no significant effect on the pelvic venous reflux. No changes in the CGRP, SP, IL-6, IL-8 and MCP-1 levels were detected after treatment. At 3 months after OVE or OVR, all patients with PeVD and combined lesions of the ovarian veins (OVs), PVs and UVs reported almost complete relief of CPP. Along with elimination of reflux in OVs, the disappearance of reflux in PVs and UVs was noted. A decrease in the CGRP and SP levels was observed (0.7 ± 0.1 ng/mL and 0.12 ± 0.02 ng/mL before treatment; 0.5 ± 0.12 ng/mL and 0.09 ± 0.06 ng/mL after treatment, respectively; all P<0.05). No changes in cytokine levels were revealed. ConclusionTreatment with VAD is associated with the CPP relief, but has no significant effect on the CGRP, SP, IL-6, IL-8, and MCP-1 levels. OVE/OVR results in the CPP relief, elimination of the pelvic venous reflux and a significant decrease in the CGRP and SP levels, but does not change cytokine levels.

STUDY ON THE INTERFERENCE OF VACCINE ANTIBODIES WITH THE SEROLOGICAL DIAGNOSIS OF LEPTOSPIROSIS IN CATTLE

A simple IgG-specific ELISA for Leptospira spp. was compared with the microscopic agglutination test (MAT) to detect IgG antibody responses to a commercial vaccine in cattle. We used an enzyme-linked immunosorbent assay (ELISA) with sonicated Leptospira interrogans serovar copenhageni M 20. After initial vaccination, specific antibodies against Leptospira spp. were detected in 90% of the animals by IgG-ELISA and 60% by MAT, while after booster, antibodies were detected in 100% and 90% of the animals by IgG-ELISA and MAT, respectively. Both serological MAT and ELISA tests revealed interferences of vaccine antibodies. Disease diagnosis with ELISA and MAT methods should be made two and a half months and four months, respectively, after vaccination to avoid interference of vaccine antibodies. On the other hand, our results suggest that IgG-ELISA may be a useful method to assess the development of IgG antibodies induced by Leptospira vaccine.

Evaluation of a Rapid Diagnostic Test (TUBEX-TF) for Diagnosis of Enteric Fever

Abstract Background and Aim: Enteric fever is a systemic febrile disease caused by Salmonella enterica serovars Typhi and Paratyphi. Culture provides a conclusive diagnosis of enteric fever. The main difficulties are a longer turnaround time and practicality in a developing nation with a huge population. The two RDTs that are now most often used are TUBEX-TF and Enterocheck-WB, where TUBEX-TF can identify IgM antibodies against S. Typhi and S. Paratyphi which are anti-O9 and anti-O2. The aim of this study was to evaluate the performance of the TUBEX-TF test in a typhoid fever endemic area of North India. Materials and Methods: In this cross-sectional study, we have performed TUBEX-TF, Enterocheck-WB /Immuno-chromatographic card test, Enzyme linked immunosorbent assay (ELISA), and Widal tests on the sera of 94 clinically suspected cases of enteric fever It was a cross-sectional comparative study that was carried out at a tertiary care institute in North India. In the present study, the ELISA test was taken as a comparative reference test. Results: ELISA, TUBEX-TF, Widal, and Immunochromatographic card test were performed on serum samples collected from 94 clinically suspected enteric fever cases, in which 50(53.19%) were positive by ELISA, 45 (47.87%) were positive by TUBEX- TF, 40 (42.5%) were positive by Typhidot and 19 (20.2%) were positive by Widal. For evaluation of TUBEX-TF, sensitivity specificity, diagnostic accuracy, positive predictive and negative predictive values, a comparison with IgM Anti-LPS ELISA test results was performed &amp; it was found 75%,79.55%,80%,74.47%, and 77.17% respectively. Conclusion: We conclude that the TUBEX-TF test has a greater sensitivity and specificity than other rapid tests currently on the market. It is a simple, rapid test to diagnose enteric fever. The necessity for large-scale research to evaluate TUBEX-TF is necessary due to interviewer variances so that it can become a novel RDT test for the diagnosis of enteric fever.

Evaluation of a Rapid Diagnostic Test (TUBEX-TF) for Diagnosis of Enteric Fever

Abstract Background and Aim: Enteric fever is a systemic febrile disease caused by Salmonella enterica serovars Typhi and Paratyphi. Culture provides a conclusive diagnosis of enteric fever. The main difficulties are a longer turnaround time and practicality in a developing nation with a huge population. The two RDTs that are now most often used are TUBEX-TF and Enterocheck-WB, where TUBEX-TF can identify IgM antibodies against S. Typhi and S. Paratyphi which are anti-O9 and anti-O2. The aim of this study was to evaluate the performance of the TUBEX-TF test in a typhoid fever endemic area of North India. Materials and Methods: In this cross-sectional study, we have performed TUBEX-TF, Enterocheck-WB /Immuno-chromatographic card test, Enzyme linked immunosorbent assay (ELISA), and Widal tests on the sera of 94 clinically suspected cases of enteric fever It was a cross-sectional comparative study that was carried out at a tertiary care institute in North India. In the present study, the ELISA test was taken as a comparative reference test. Results: ELISA, TUBEX-TF, Widal, and Immunochromatographic card test were performed on serum samples collected from 94 clinically suspected enteric fever cases, in which 50(53.19%) were positive by ELISA, 45 (47.87%) were positive by TUBEX- TF, 40 (42.5%) were positive by Typhidot and 19 (20.2%) were positive by Widal. For evaluation of TUBEX-TF, sensitivity specificity, diagnostic accuracy, positive predictive and negative predictive values, a comparison with IgM Anti-LPS ELISA test results was performed &amp; it was found 75%,79.55%,80%,74.47%, and 77.17% respectively. Conclusion: We conclude that the TUBEX-TF test has a greater sensitivity and specificity than other rapid tests currently on the market. It is a simple, rapid test to diagnose enteric fever. The necessity for large-scale research to evaluate TUBEX-TF is necessary due to interviewer variances so that it can become a novel RDT test for the diagnosis of enteric fever.

Comparative Study of Various Diagnostic Methods for Detection of Mycoplasma gallisepticum and Mycoplasma synoviae in Egyptian Chicken Flocks

The significance of Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) in the poultry industry underscores the critical importance of diagnosing avian mycoplasmosis within the field of veterinary medicine. The present study aimed to compare various diagnostic methods for detecting MG and MS in Egyptian Chicken Flocks. A total of 360 samples were collected from breeder, layer, and broiler chickens from four governorates in Egypt. Conventional isolation methods and polymerase chain reaction (PCR) were used for the direct detection of MG and MS, while serum plate agglutination test (SPA) and Enzyme-linked immunosorbent assay (ELISA) were used for detecting antibodies against MG and MS. The highest detection rate of MG was found in commercial layers, followed by breeders, and broilers. Regarding MS, the highest detection rate was found in breeders, followed by commercial layers, and broilers. By comparing the used diagnostic methods, MG and MS were determined by the SPA test (40% and 31.1% respectively), ELISA test (31.7% and 23.6%), PCR (16.7% and 11.7%), and by the conventional culture method (10.8% and 3.9%). It could be concluded that the serological methods and PCR gave better sensitivity than culture methods and can be used in the diagnosis of avian mycoplasmosis.

Application of PCR and ELISA for Investigation the Helicobacter pylori in Stool Specimens of Infected Patients

Background: Infection with Helicobacter pylori is a global health issue, and non-invasive and accurate testing is a promising diagnostic method. Because H. pylori antibodies collapse quite gradually even after effective treatment. So, the tests for identifying the antibody of H. pylori are missing specificity and sensitivity. Instead, the tests of stool antigen for H. pylori are described as an alternative method because they have more reliability and simplicity. Aim: The study was aimed to assess the performance of enzyme-linked immunosorbent assays (ELISA), for the detection of H. pylori antigen in stool and compare it to molecular tests by using two housekeeping genes. Methods: Stool samples were collected from patients (85) who attended the Gastroenterology and Hepatology Teaching Center in Baghdad, Iraq, and underwent esophagogastroduodenoscopy (EGD) for biopsy in a period extended from November 2020 to July 2021. PCR and ELISA were used to Investigate H. pylori in stool specimens of infected patients. Results: The result revealed that 74/85 (87.88%) of specimens were positive by ELISA, whereas 49/85 (57.6%) were positive for the 16 SrRNA gene related to Helicobacter genus and 35/85 (41.1%) were positive for amiA gene related to H. pylori species, and that means the ELISA test is more accurate than PCR analysis for detection of H. pylori bacteria in stool specimen. Conclusion: Detection H. pylori Ag using the ELISA test was superior to the molecular test. Therefore, the ELISA test might replace other methods and could be used for the detection of active H. pylori infection before the inauguration of treatment between dyspeptic patients.

Application of PCR and ELISA for Investigation the Helicobacter pylori in Stool Specimens of Infected Patients

Background: Infection with Helicobacter pylori is a global health issue, and non-invasive and accurate testing is a promising diagnostic method. Because H. pylori antibodies collapse quite gradually even after effective treatment. So, the tests for identifying the antibody of H. pylori are missing specificity and sensitivity. Instead, the tests of stool antigen for H. pylori are described as an alternative method because they have more reliability and simplicity. Aim: The study was aimed to assess the performance of enzyme-linked immunosorbent assays (ELISA), for the detection of H. pylori antigen in stool and compare it to molecular tests by using two housekeeping genes. Methods: Stool samples were collected from patients (85) who attended the Gastroenterology and Hepatology Teaching Center in Baghdad, Iraq, and underwent esophagogastroduodenoscopy (EGD) for biopsy in a period extended from November 2020 to July 2021. PCR and ELISA were used to Investigate H. pylori in stool specimens of infected patients. Results: The result revealed that 74/85 (87.88%) of specimens were positive by ELISA, whereas 49/85 (57.6%) were positive for the 16 SrRNA gene related to Helicobacter genus and 35/85 (41.1%) were positive for amiA gene related to H. pylori species, and that means the ELISA test is more accurate than PCR analysis for detection of H. pylori bacteria in stool specimen. Conclusion: Detection H. pylori Ag using the ELISA test was superior to the molecular test. Therefore, the ELISA test might replace other methods and could be used for the detection of active H. pylori infection before the inauguration of treatment between dyspeptic patients.

Wind disturbance-based tomato seedlings growth control

Wind-disturbance is a potential eco-friendly technique for tackling leggy seedlings. This study uses orthogonal experimental design and seedlings vigour assessment by strong seedling index (SSI) to investigate wind-disturbance on regulating tomato seedlings growth. Changes in endogenous hormone levels and biomechanical properties of tomato seedlings were investigated using enzyme-linked immunosorbent assay and uniaxial tension tests. Results showed that factors influencing significantly SSI, in descending order, are wind disturbance time (T), seedling age at the onset of wind disturbance (SA), wind velocity (V), and interval time (I). The wind-disturbance effect was found to be optimal with SSI = 0.126 for a condition where V, T, I and SA values are 3 m s−1, 1 min, 30 min, and 15 days, respectively. V and T were positively correlated with the ethylene and abscisic acid content in seedling leaves, abscisic acid and auxin content in stems, cytokinin and ethylene content in roots, and the elastic moduli of stems and roots but negatively associated with the cytokinin content in stems and leaves, auxin and abscisic acid content in roots, and leaves’ elastic modulus. Wind disturbance mechanism for controlling seedlings growth involved eliciting the accumulation of abscisic acid in stems and leaves and reduction of the auxin content in roots to about the optimal threshold for roots growth thereby reducing seedling stems and leaves development and promoting a better roots growth and a high SSI. This work offers theoretical insights and technical guidance for utilising wind-disturbance as a sustainable seedling cultivation and personalised seedling management approach.

Dengue Virus and Zika Virus Seroprevalence in the South Pacific Populations of the Cook Islands and Vanuatu

Arboviral diseases are serious threats to global health with increasing prevalence and potentially severe complications. Significant arthropod-borne viruses are the dengue viruses (DENV 1-4), the Zika virus (ZIKV), and the chikungunya virus (CHIKV). Among the areas most affected is the South Pacific Region (SPR). Here, arboviruses not only cause a high local burden of disease, but the region has also proven to contribute to their global spread. Outpatient serum samples collected between 08/2016 and 04/2017 on three islands of the island states of Vanuatu and the Cook Islands were tested for anti-DENV- and anti-ZIKV-specific antibodies (IgG) using enzyme-linked immunosorbent assays (ELISA). ELISA test results showed 89% of all test sera from the Cook Islands and 85% of the Vanuatu samples to be positive for anti-DENV-specific antibodies. Anti-ZIKV antibodies were identified in 66% and 52%, respectively, of the test populations. Statistically significant differences in standardized immunity levels were found only at the intranational level. Our results show that in both the Cook Islands and Vanuatu, residents were exposed to significant Flavivirus transmission. Compared to other seroprevalence studies, the marked difference between ZIKV immunity levels and previously published CHIKV seroprevalence rates in our study populations is surprising. We propose the timing of ZIKV and CHIKV emergence in relation to recurrent DENV outbreaks and the impact of seasonality as explanatory external factors for this observation. Our data add to the knowledge of arboviral epidemics in the SPR and contribute to a better understanding of virus spread, including external conditions with potential influence on outbreak dynamics. These data may support preventive and rapid response measures in the affected areas, travel-related risk assessment, and infection identification in locals and returning travelers.

METTL5 promotes gastric cancer progression via sphingomyelin metabolism.

The treatment of gastric cancer (GC) has caused an enormous social burden worldwide. Accumulating studies have reported that N6-methyladenosine (m6A) is closely related to tumor progression. METTL5 is a m6A methyltransferase that plays a pivotal role in maintaining the metabolic stability of cells. However, its aberrant regulation in GC has not been fully elucidated. To excavate the role of METTL5 in the development of GC. METTL5 expression and clinicopathological characteristics were analyzed via The Cancer Genome Atlas dataset and further verified via immunohistochemistry, western blotting and real-time quantitative polymerase chain reaction in tissue microarrays and clinical samples. The tumor-promoting effect of METTL5 on HGC-27 and AGS cells was explored in vitro by Cell Counting Kit-8 assays, colony formation assays, scratch healing assays, transwell assays and flow cytometry. The tumor-promoting role of METTL5 in vivo was evaluated in a xenograft tumor model. The EpiQuik m6A RNA Methylation Quantification Kit was used for m6A quantification. Next, liquid chromatography-mass spectrometry was used to evaluate the association between METTL5 and sphingomyelin metabolism, which was confirmed by Enzyme-linked immunosorbent assay and rescue tests. In addition, we investigated whether METTL5 affects the sensitivity of GC cells to cisplatin via colony formation and transwell experiments. Our research revealed substantial upregulation of METTL5, which suggested a poor prognosis of GC patients. Increased METTL5 expression indicated distant lymph node metastasis, advanced cancer stage and pathological grade. An increased level of METTL5 correlated with a high degree of m6A methylation. METTL5 markedly promotes the proliferation, migration, and invasion of GC cells in vitro. METTL5 also promotes the growth of GC in animal models. METTL5 knockdown resulted in significant changes in sphingomyelin metabolism, which implies that METTL5 may impact the development of GC via sphingomyelin metabolism. In addition, high METTL5 expression led to cisplatin resistance. METTL5 was found to be an oncogenic driver of GC and may be a new target for therapy since it facilitates GC carcinogenesis through sphingomyelin metabolism and cisplatin resistance.

Acteoside and isoacteoside alleviate renal dysfunction and inflammation in lipopolysaccharide-induced acute kidney injuries through inhibition of NF-κB signaling pathway.

Acute kidney injury (AKI) is a sudden loss of renal function with a high mortality rate and inflammation is thought to be the underlying cause. The phenylpropanoid components acteoside (ACT) and isoacteoside (ISO), which were isolated from Cistanche deserticola Y.C.Ma, have been reported to have preventive effects against kidney disorders. This study aimed to investigate the anti-inflammatory properties and protective mechanisms of ACT and ISO. In this investigation, kidney function was assessed using a semi-automatic biochemical analyzer, histopathology was examined using Hematoxylin-Eosin staining and immunohistochemistry, and the concentration of inflammatory cytokines was assessed using an enzyme-linked immunosorbent assay (ELISA) test. In addition, using Western blot and q-PCR, the expression of proteins and genes connected to the NF-κB signaling pathway in mice with lipopolysaccharide (LPS)-induced AKI was found. The findings showed that under AKI intervention in LPS group, ACT group and ISO group, the expression of Rela (Rela gene is responsible for the expression of NFκB p65 protein) and Tlr4 mRNA was considerably elevated (P<0.01), which led to a significant improvement in the expression of MyD88, TLR4, Iκ-Bɑ and NF-κB p65 protein (P<0.001). The levels of Alb, Crea and BUN (P<0.001) increased along with the release of downstream inflammatory factors such as IL-1β, IL-6, Cys-C, SOD1 and TNF-α (P<0.001). More importantly, the study showed that ISO had a more favorable impact on LPS-induced AKI mice than ACT. In conclusion, by inhibiting NF-κB signaling pathway, ACT and ISO could relieve renal failure and inflammation in AKI, offering a fresh possibility for the therapeutic management of the condition.

Nine-year seroepidemiological study of severe fever with thrombocytopenia syndrome virus infection in feral horses in Cape Toi, Japan

Severe fever with thrombocytopenia syndrome (SFTS) is a fatal zoonosis caused by ticks in East Asia. As SFTS virus (SFTSV) is maintained between wildlife and ticks, seroepidemiological studies in wildlife are important to understand the behavior of SFTSV in the environment. Miyazaki Prefecture, Japan, is an SFTS-endemic area, and approximately 100 feral horses, called Misaki horses (Equus caballus), inhabit Cape Toi in Miyazaki Prefecture. While these animals are managed in a wild-like manner, their ages are ascertainable due to individual identification. In the present study, we conducted a seroepidemiological survey of SFTSV in Misaki horses between 2015 and 2023. This study aimed to understand SFTSV infection in horses and its transmission to wildlife. A total of 707 samples from 180 feral horses were used to determine the seroprevalence of SFTSV using enzyme-linked immunosorbent assay (ELISA). Neutralization testing was performed on 118 samples. In addition, SFTS viral RNA was detected in ticks from Cape Toi and feral horses. The overall seroprevalence between 2015 and 2023 was 78.5% (555/707). The lowest seroprevalence was 55% (44/80) in 2016 and the highest was 92% (76/83) in 2018. Seroprevalence was significantly affected by age, with 11% (8/71) in those less than one year of age and 96.7% (435/450) in those four years of age and older (p < 0.0001). The concordance between ELISA and neutralization test results was 88.9% (105/118). SFTS viral RNA was not detected in ticks (n = 516) or feral horses. This study demonstrated that horses can be infected with SFTSV and that age is a significant factor in seroprevalence in wildlife. This study provides insights into SFTSV infection not only in horses but also in wildlife in SFTS-endemic areas.

Interleukin levels and depressive symptoms in psoriatic arthritis patients: insights from a case-control study on socio-demographic factors and disease perception.

In the course of psoriatic arthritis (PsA), depression occurs much more often than in the general population. Depression can be considered a poor prognostic factor. The aim of the study was to assess the relationships between the occurrence of depression and the levels of proinflammatory cytokines in patients with PsA. The study included 86 (47F/39M) patients with PsA. Only patients with high disease activity (DAPSA > 28) were enrolled in the study. The severity of depressive symptoms was assessed using the Beck Depression Inventory II (BDI-II) for all patients. Additionally, sociodemographic data were collected. All patients were also assessed for the levels of interleukins (IL): IL-1, IL-6, IL-17A, IL-23, and tumor necrosis factor alpha (TNF-α) using the enzyme-linked immunosorbent assay (ELISA) test. In the study group, depression (BDI-II ≥ 14) was diagnosed in 45 patients (52%). Patients with coexisting depression reported higher levels of pain and disease activity on the visual analogue scale compared to patients without depression (8.5 vs. 7.7, p < 0.001 and 9.3 vs. 8.4, p < 0.001, respectively). The mean levels of proinflammatory cytokines [pg/ml], IL-1 and IL-6, were also higher in the group of patients with depression (46.4 vs. 4.7, p < 0.001 and 10.5 vs. 4.9, p < 0.001, respectively). The coexistence of depression in the course of Psoriatic Arthritis (PsA) is associated with higher levels of IL-1 and IL-6. Depression has a negative impact on the perception of the underlying disease and is linked to reduced social and occupational activity.

Seroprevalence of Helicobacter pylori IgG Antibodies and Associated Risk Factors among Asymptomatic Vegetable Sellers in Uyo, South Nigeria

Background: Vegetables are essential part of people’s diet and are sometimes consumed raw and often without heat treatment or thorough washing, hence some studies suggested it serves as vehicle for the transmission of pathogenic microorganisms like Helicobacter pylori. The source of contamination has also been traced to sellers of vegetables who are involved in unhygienic practices which contribute to the dissemination of pathogenic microorganisms. Therefore the aim of this study was to provide information on the seroprevalence of H. pylori IgG antibodies and associated risk factors among asymptomatic vegetable sellers in Uyo, Akwa Ibom State, Nigeria. Materials and Methods: Blood and stool samples from 142 participants were included in the study. Blood samples were analyzed using the rapid diagnostic method while the stool samples were analyzed using Enzyme linked immunosorbent Assays (ELISA). Results: A total of 85 (59.9%) were positive by rapid diagnostic test (RDT) while 53 (37.3%) were positive by stool ELISA test. Risk factors like age, marital status, residency family size, years in vegetable business, source of vegetable, wash before stalling etc were all statistically associated with (p&lt;0.05) with H. pylori prevalence. Conclusion: This study revealed that unhygienic behaviors by vegetable sellers are associated with high risk of H. pylori infection to vegetable consumers.

An indirect ELISA for detecting anti-SARS-CoV-2 antibodies in human sera using a baculovirus-expressed recombinant nucleocapsid antigen

This study focuses on the development and initial assessment of an indirect IgG enzyme-linked immunosorbent assay (ELISA) specifically designed to detect of anti-SARS-CoV-2 antibodies. The unique aspect of this ELISA method lies in its utilization of a recombinant nucleocapsid (N) antigen, produced through baculovirus expression in insect cells. Our analysis involved 292 RT-qPCR confirmed positive serum samples and 54 pre-pandemic healthy controls. The process encompassed cloning, expression, and purification of the SARS-CoV-2 N gene in insect cells, with the resulted purified protein employed in our ELISA tests. Statistical analysis yielded an Area Under the Curve of 0.979, and the optimized cut-off exhibited 92 % sensitivity and 94 % specificity. These results highlight the ELISA's potential for robust and reliable serological detection of SARS-CoV-2 antibodies. Further assessments, including a larger panel size, reproducibility tests, and application in diverse populations, could enhance its utility as a valuable biotechnological solution for diseases surveillance.

A Mini-Review on Elisa-Based Diagnosis of Schistosomiasis.

Schistosomiasis is a neglected tropical parasitic disease caused by trematode worms of the genus schistosoma, which affects approximately 240 million people worldwide. the diagnosis of the disease can be performed by parasitological, molecular, and/or immunological methods, however, the development of new diagnostic methods still essential to guide policy decisions, monitor disease trends and assess the effectiveness of interventions. in this sense, the current work summarizes the findings of a systematic review regarding antigens applied in the enzyme-linked immunosorbent assay test, which were patented and published over the last ten years. the literature search strategy used medical subject heading (mesh) terms to define as descriptors. "schistosoma mansoni" was used in arrangement with the descriptors "immunoassay", "enzyme-linked immunosorbent assay", "elisa", and "antigens", using the "and" connector. the patent search was done using keywords, including diagnosis and schistosoma or schistosomiasis or schistosome. several databases were employed for the patent search, such as intellectual property national institute; european patent office; the united states patent and trademark office; patent scope, and google patents. forty-one articles were retrieved, of which only five met the eligibility criteria. seventeen patents were taken from the databases, and a brief description of the most relevant inventions is given here. schistosomiasis is considered the most important helminthic disease in worldwide. therefore, it is important to of searching for and develops diagnostic methods based on serology to reduce morbidity and mortality caused by the disease.

A bispecific antibody targeting HLA-DQ2.5-gluten peptides potently blocks gluten-specific T cells induced by gluten ingestion in patients with celiac disease

The gluten-free diet for celiac disease (CeD) is restrictive and often fails to induce complete symptom and/or mucosal disease remission. Central to CeD pathogenesis is the gluten-specific CD4+ T cell that is restricted by HLA-DQ2.5 in over 85% of CeD patients, making HLA-DQ2.5 an attractive target for suppressing gluten-dependent immunity. Recently, a novel anti-HLA-DQ2.5 antibody that specifically recognizes the complexes of HLA-DQ2.5 and multiple gluten epitopes was developed (DONQ52). ObjectiveTo assess the ability of DONQ52 to inhibit CeD patient-derived T-cell responses to the most immunogenic gluten peptides that encompass immunodominant T cell epitopes. MethodsWe employed an in vivo gluten challenge model in patients with CeD that affords a quantitative readout of disease-relevant gluten-specific T-cell responses. HLA-DQ2.5+ CeD patients consumed food containing wheat, barley, or rye for 3 days with collection of blood before (D1) and 6 days after (D6) commencing the challenge. Peripheral blood mononuclear cells were isolated and assessed in an interferon (IFN)-γ enzyme-linked immunosorbent spot assay (ELISpot) testing responses to gluten peptides encompassing a series of immunodominant T cell epitopes. The inhibitory effect of DONQ52 (4 or 40 μg/mL) was assessed and compared to pan-HLA-DQ blockade (SPVL3 antibody). ResultsIn HLA-DQ2.5+ CeD patients, DONQ52 reduced T cell responses to all wheat gluten peptides to an equivalent or more effective degree than pan-HLA-DQ antibody blockade. It reduced T cell responses to a cocktail of the most immunodominant wheat epitopes by a median of 87% (IQR 72–92). Notably, DONQ52 also substantially reduced T-cell responses to dominant barley hordein and rye secalin derived peptides. DONQ52 had no effect on T-cell responses to non-gluten antigens. ConclusionDONQ52 can significantly block HLA-DQ2.5-restricted T cell responses to the most highly immunogenic gluten peptides in CeD. Our findings support in vitro data that DONQ52 displays selectivity and broad cross-reactivity against multiple gluten peptide:HLA-DQ2.5 complexes. This work provides proof-of-concept multi-specific antibody blockade has the potential to meaningfully inhibit pathogenic gluten-specific T-cell responses in CeD and supports ongoing therapeutic development.

Electronic alerts to improve management of heparin-induced thrombocytopenia

BackgroundHeparin-induced thrombocytopenia (HIT) is a difficult clinicopathologic diagnosis to make and to treat. Delays in identification and appropriate treatment can lead to increased morbidity and mortality. ObjectivesTo use electronic health alert interventions to improve provider diagnosis and management of heparin-induced thrombocytopenia through guideline-based, accurate care delivery. MethodsThis quality improvement initiative developed 3 electronic health record-based interventions at our 750-bed academic medical center to improve the initial management of suspected HIT between 2018 and 2021: 1. an interruptive alert to recommend discontinuation of active heparin products when signing a heparin-platelet factor 4 test (PF4) order, 2. integrated 4T score calculation in the heparin-PF4 test order, and 3. interruptive alert suggesting not to order heparin-PF4 tests when the 4T score is <4. Changes in practice were assessed over defined time periods pre and post each intervention. ResultsIntervention 1 resulted in heparin discontinuation in more patients, with 65% (191 heparin orders/293 heparin-PF4 enzyme-linked immunosorbent assay tests) of cases continuing heparin prealert and only 54% (127 heparin orders/235 heparin-PF4 enzyme-linked immunosorbent assay tests) postinterruptive alert (95% CI 2.3-19.9; P = .015). Intervention 2 increased appropriate heparin-PF4 test ordering from 40.4% (110/272) preintervention to 79.1% (246/311) (95% CI 30.9-46.4; P < .00001) postintervention, with inappropriate PF4 ordering defined as testing when 4T score was <4. Intervention 3 did not lead to reduction in heparin-PF4 testing in the control group (96 inappropriate orders/402 total orders, 24%) compared to the randomized alert group (56 inappropriate orders/298 total orders; 19%) (95% CI −1.2 to 11.5; P = .13). ConclusionImplementation of unique electronic health record interventions, including both diagnostic and management interventions, led to improved guideline-based, accurate care delivery with 4T score calculation and cessation of heparin for patients with suspected HIT.

Validation of an enzyme-linked immunosorbent assay test-system for bevacizumab concentration determing in biological fluids

Introduction. One of the reasons for the development of many diseases primarily malignant is the increased expression of Vascular Endothelial Growth Factor (VEGF). Bevacizumab is a drug that neutralizes the biological activity of VEGF. The molecular structure of bevacizumab is a recombinant humanized antibody. Its use reduces vascularization in the foci of increased VEGF expression which slows down tumor growth and also helps restore vision in a number of ophthalmic diseases. To determine the concentration of bevacizumab in human biological fluids a test system based on Enzyme-linked Immunosorbent Assay (ELISA) is presented.Aim. Aim of this study is the validation of this test system.Materials and methods. Blank sera of volunteers, bevacizumab solution, enzyme-linked immunosorbent assay, solid-phase sandwich ELISA kit, microplate photometer.Results and discussion. The following characteristics of the test system were determined: the lower limit of quantification is 2.0 mcg/ml, the upper calibration range is up to 200 mcg/ml, the accuracy and precision within one series and between series does not exceed 20 %, and the total error of the method – 30 %, short-term stability for samples at room temperature – 6 hours, long-term stability – 14 days at –20 °C, the ability to freeze/thaw of samples is up to three times, the ability to determine samples with a concentration above the upper calibrator after diluting in 2 times.Conclusion. The results obtained fully comply with international acceptance criteria and allow the use of the ELISA test system manufactured by LLC "Probiotek" for use in the field of clinical laboratory diagnostics.