scholarly journals Development of an Indirect ELISA Based on Spike Protein to Detect Antibodies against Feline Coronavirus

Viruses ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 2496
Author(s):  
Bo Dong ◽  
Gaoqiang Zhang ◽  
Xiaodong Zhang ◽  
Xufei Chen ◽  
Meiling Zhang ◽  
...  
Keyword(s):  
Operating Characteristic ◽  
Characteristic Curve ◽  
Systemic Disease ◽  
Analysis Data ◽  
Spike Protein ◽  
Diagnostic Tools ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Pathogenic Virus ◽  
Feline Coronavirus

Feline coronavirus (FCoV) is a pathogenic virus commonly found in cats that causes a benign enteric illness and fatal systemic disease, feline infectious peritonitis. The development of serological diagnostic tools for FCoV is helpful for clinical diagnosis and epidemiological investigation. Therefore, this study aimed to develop an indirect enzyme-linked immunosorbent assay (iELISA) to detect antibodies against FCoV using histidine-tagged recombinant spike protein. FCoV S protein (1127–1400 aa) was expressed and used as an antigen to establish an ELISA. Mice and rabbits immunized with the protein produced antibodies that were recognized and bound to the protein. The intra-assay coefficient of variation (CV) was 1.15–5.04% and the inter-assay CV was 4.28–15.13%, suggesting an acceptable repeatability. iELISA did not cross-react with antisera against other feline viruses. The receiver operating characteristic curve analysis revealed an 86.7% sensitivity and 93.3% specificity for iELISA. Serum samples (n = 107) were tested for anti-FCoV antibodies, and 70.09% of samples were positive for antibodies against FCoV. The iELISA developed in our study can be used to measure serum FCoV antibodies due to its acceptable repeatability, sensitivity, and specificity. Additionally, field sample analysis data demonstrated that FCoV is highly prevalent in cat populations in Fujian province, China.

scholarly journals Prevalence of feline coronavirus antibodies in cats in Bursa province, Turkey, by an enzyme-linked immunosorbent assay

2009 ◽  
Vol 11 (10) ◽  
pp. 881-884 ◽  
Author(s):  
Annamaria Pratelli ◽  
Kadir Yesilbag ◽  
Marcello Siniscalchi ◽  
Ebru Yalçm ◽  
Zeki Yilmaz
Keyword(s):  
Immunosorbent Assay ◽  
Predictive Value ◽  
Gold Standard ◽  
Population Based ◽  
Standard Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
High Association ◽  
Gold Standard Test ◽  
Feline Coronavirus

Feline sera from Bursa province (Turkey) were assayed for coronavirus antibody using an enzyme-linked immunosorbent assay (ELISA). The study was performed on 100 sera collected from cats belonging to catteries or community shelters and to households. The serum samples were initially tested with the virus neutralisation (VN) test and the results were then compared with the ELISA. The VN yielded 79 negative and 21 positive sera but the ELISA confirmed only 74 as negative. The ELISA-negative sera were also found to be free of feline coronoviruses-specific antibodies by Western blotting. Using the VN as the gold standard test, ELISA had a sensitivity of 100% and a specificity of 93.6%, with an overall agreement of 95%. The Kappa (κ) test indicated high association between the two tests (κ=0.86, 95% confidence interval (CI) 0.743–0.980). The positive predictive value (PPV) was 0.8, and the negative predictive value (NPV) was 0.93. The prevalence of FCoV II antibodies in the sampled population based on the gold standard was 62% (95% CI 0.44–0.77) among multi-cat environments, and 4% (95% CI 0.01–0.11) among single cat households.

scholarly journals Different expression pattern of human cytomegalovirus-encoded microRNAs in circulation from virus latency to reactivation

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Wanqing Zhou ◽  
Cheng Wang ◽  
Meng Ding ◽  
Yuying Bian ◽  
Yujie Zhong ◽  
...  
Keyword(s):  
Human Cytomegalovirus ◽  
Characteristic Curve ◽  
Antiviral Treatment ◽  
Serum Levels ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Prediction Ability ◽  
Stem Loop ◽  
Virus Latency ◽  
Polymerase Chain

Abstract Background Human cytomegalovirus (HCMV) is a beta-hersvirinae that has a high latent infection rate worldwide and can cause serious consequences in immunocompromised patients when reactivation; however, the mechanism of how HCMV convert from latent to reactivation has rarely been investigated. In the present study, we aimed to perform a comprehensive analysis of the HCMV-encoded microRNA (miRNA) profile in serum of patients upon HCMV reactivation from latency and to further evaluate its clinical significance for the disease monitoring and preventing usefulness. Methods Serum samples from 59 viremia patients and 60 age-gender matched controls were enrolled in this study for screening and validation of different expression of HCMV miRNAs. Serum concentrations of 22 known HCMV miRNAs were determined by a hydrolysis probe-based stem-loop quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay. HCMV DNA was measured by quantitative real-time PCR (qPCR) with the whole blood sample. Serum HCMV IgG and IgM were assessed using enzyme linked immunosorbent assay (ELISA). Another 47 samples from 5 patients at different time points were collected to evaluate the monitoring effectiveness and disease prediction ability of differential expression HCMV-miRNAs during the antiviral treatment. Results The RT-qPCR analysis revealed that the serum levels of 16 of the 22 examined HCMV miRNAs were elevated in HCMV viremia patients compared with controls, and a profile of 8 HCMV miRNAs including hcmv-miR-US25-2-3p, hcmv-miR-US4-5p, hcmv-miR-US25-2-5p, hcmv-miR-US25-1-3p, hcmv-miR-US25-1, hcmv-miR-UL36, hcmv-miR-UL148D, hcmv-miR-US29-3p were markedly elevated (fold change > 2, P < 0.01). Receiver operating characteristic curve (ROC) analysis were performed on the selected HCMV-miRNAs in all of the patients and controls that enrolled in this study, and which ranged from 0.72 to 0.80 in the autoimmune patients. In addition, hcmv-miR-US25-1-3p levels were significantly correlated with HCMV DNA load (r = 0.349, P = 0.007), and were obviously higher in the reactivation set than the latency set in the autoimmune patients, which could be a predictor for the monitoring of the antiviral treatment. Conclusions HCMV miRNAs profile showed markedly shift-switch from latency to reactivation in circulation from HCMV infected patients and hcmv-miR-US25-1-3p may be served as a predictor for the switch upon reactivation from latency in patients suffered with autoimmune diseases.

scholarly journals Evaluation of a Commercial Enzyme-Linked Immunosorbent Assay for the Diagnosis of Paratuberculosis in Dairy Cattle

1995 ◽  
Vol 7 (4) ◽  
pp. 488-493 ◽  
Author(s):  
Raymond W. Sweeney ◽  
Robert H. Whitlock ◽  
Carol L. Buckley ◽  
Pam A. Spencer
Keyword(s):  
Receiver Operating Characteristic Curve ◽  
Receiver Operating Characteristic ◽  
Operating Characteristic ◽  
Characteristic Curve ◽  
Curve Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Improve Performance ◽  
Operating Characteristic Curve ◽  
Elisa Result ◽  
Clinical Cases

The performance of a commercially available ELISA for detection of antibodies to Mycobacterium paratuberculosis was evaluated using sera from 1,146 cows. Samples were from uninfected cattle, infected subclinical cattle shedding low numbers of organism in feces, subclinical heavy shedders, clinical cases, and randomly selected cattle in a slaughterhouse survey for paratuberculosis. The overall sensitivity of the test, using the manufacturer's recommended cutoff was 45% ± 4.8%, and the specificity was 99% ± 0.9%. The ELISA result was significantly correlated with the number of colonies of M. paratuberculosis detected by fecal culturing. The sensitivity of the test was highest for clinical cases of paratuberculosis (87% ± 8.4%), and lowest for subclinical, light-shedding cattle (15% ± 6.6%). Changing the cutoff point did not improve performance of the test. Evaluating ELISA results with a kinetics-based method reduced plate-to-plate variation in results but did not improve performance of the test based on receiver-operating characteristic curve analysis.

scholarly journals Calprotectin is a potential prognostic marker for polycystic ovary syndrome

2016 ◽  
Vol 54 (2) ◽  
pp. 253-257
Author(s):  
Shouzhen Chen ◽  
Mei Jiang ◽  
Tao Ding ◽  
Junmei Wang ◽  
Ping Long
Keyword(s):  
Insulin Resistance ◽  
Polycystic Ovary Syndrome ◽  
Receiver Operating Characteristic ◽  
Operating Characteristic ◽  
Polycystic Ovary ◽  
Characteristic Curve ◽  
Enzyme Linked Immunosorbent Assay ◽  
Ovary Syndrome ◽  
Inflammatory Conditions ◽  
Receiver Operating

Background Calprotectin is an antimicrobial, calcium and zinc-binding heterocomplex protein and has been proposed as a marker to rule out inflammatory conditions. The aim of this study was to evaluate the role of calprotectin in the diagnosis of polycystic ovary syndrome and to investigate the association between calprotectin and insulin resistance. Methods A total of 41 females with polycystic ovary syndrome and 54 age-matched without polycystic ovary syndrome were eligible for the study. Serum concentration of calprotectin was determined using enzyme-linked immunosorbent assay. Clinical characteristics, hormone and metabolic parameters were evaluated in each subject. The predictive value of serum calprotectin was assessed using receiver operating characteristic curves. Correlations between the serum calprotectin concentrations and insulin resistance were examined using Spearman’s correlation. Results We found that the serum calprotectin concentrations were significantly higher in polycystic ovary syndrome compared with the non-polycystic ovary syndrome group ( P < 0.001). The area under the receiver operating characteristic curve assay yielded a satisfactory result of 0.88 (95% confidence interval 0.81–0.95; P < 0.001). The optimum cut-off was 2.4  µg/mL with a 85.2% specificity and 75.6% sensitivity for polycystic ovary syndrome diagnosis. A significant positive correlation was found between the serum calprotectin and insulin resistance. Conclusions These results suggest that calprotectin might be a useful adjunct in the diagnosis of polycystic ovary syndrome, especially those with insulin resistance.

scholarly journals Immunodiagnosis of cattle fascioliasis using a 27 kDa Fasciola gigantica antigen

2021 ◽  
pp. 2097-2101
Author(s):  
Mohamed J. Saadh ◽  
Samer A. Tanash ◽  
Ammar M. Almaaytah ◽  
Issam J. Sa'adeh ◽  
Saed M. Aldalaen ◽  
...  
Keyword(s):  
Western Blotting ◽  
Clinical Symptoms ◽  
Characteristic Curve ◽  
Fasciola Gigantica ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infected Cattle ◽  
Serum Samples ◽  
Healthy Cattle ◽  
Target Epitope ◽  
Circulating Antigens

Background and Aim: Diagnosis of fascioliasis depends on clinical symptoms and routine laboratory tests. Recently, antibodies and circulating antigens of Fasciola were used for detecting active infections. Therefore, this study aimed to identify Fasciola gigantica antigens in the sera of infected cattle using Western blotting and enzyme-linked immunosorbent assay (ELISA) for an accurate diagnosis of cattle infected with F. gigantica. Materials and Methods: Serum samples were obtained from 108, 23, and 19 cattle infected with Fasciola gigantica, Paramphistomum cervi, and Strongylids, respectively, including 57 non-infected cattle that were used as healthy cattle for the study. Western blotting and ELISA were then used to detect circulating Fasciola antigens at 27 kDa. Results: The target epitope was detected in an F. gigantica adult-worm antigen preparation, excretory/secretory products, and serum from cattle infected with F. gigantica. However, it was absent in sera from P. cervi, Strongylids, and healthy cattle. The purified 27 kDa F. gigantica (FPA-27) antigen was also detected in cattle serum using ELISA with high degrees of sensitivity and specificity (94% and 82%, respectively), and the area under the receiver operating characteristic curve was 0.89 with a highly significant correlation of p<0.0001. Conclusion: The FPA-27 is proposed to be a promising candidate for the serodiagnosis of fascioliasis in cattle.

scholarly journals Development and application of a fluorescence protein microarray for detecting serum alpha-fetoprotein in patients with hepatocellular carcinoma

2016 ◽  
Vol 44 (6) ◽  
pp. 1414-1423 ◽  
Author(s):  
Aiying Zhang ◽  
Chengzeng Yin ◽  
Zhenshun Wang ◽  
Yonghong Zhang ◽  
Yuanshun Zhao ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Protein Microarray ◽  
Characteristic Curve ◽  
Alpha Fetoprotein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Control Subjects ◽  
Serum Alpha Fetoprotein ◽  
Healthy Control ◽  
Fluorescence Protein

Objective To develop a simple, effective, time-saving and low-cost fluorescence protein microarray method for detecting serum alpha-fetoprotein (AFP) in patients with hepatocellular carcinoma (HCC). Method Non-contact piezoelectric print techniques were applied to fluorescence protein microarray to reduce the cost of prey antibody. Serum samples from patients with HCC and healthy control subjects were collected and evaluated for the presence of AFP using a novel fluorescence protein microarray. To validate the fluorescence protein microarray, serum samples were tested for AFP using an enzyme-linked immunosorbent assay (ELISA). Results A total of 110 serum samples from patients with HCC ( n = 65) and healthy control subjects ( n = 45) were analysed. When the AFP cut-off value was set at 20 ng/ml, the fluorescence protein microarray had a sensitivity of 91.67% and a specificity of 93.24% for detecting serum AFP. Serum AFP quantified via fluorescence protein microarray had a similar diagnostic performance compared with ELISA in distinguishing patients with HCC from healthy control subjects (area under receiver operating characteristic curve: 0.906 for fluorescence protein microarray; 0.880 for ELISA). Conclusion A fluorescence protein microarray method was developed for detecting serum AFP in patients with HCC.

scholarly journals IgGFc-binding protein in pregnancies complicated by spontaneous preterm delivery: a retrospective cohort study

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jaroslav Stranik ◽  
Marian Kacerovsky ◽  
Ondrej Soucek ◽  
Martina Kolackova ◽  
Ivana Musilova ◽  
...  
Keyword(s):  
Amniotic Fluid ◽  
Preterm Labor ◽  
Operating Characteristic ◽  
Binding Protein ◽  
Characteristic Curve ◽  
Enzyme Linked Immunosorbent Assay ◽  
Amniotic Cavity ◽  
Spontaneous Preterm Delivery ◽  
Microbial Invasion ◽  
Prelabor Rupture Of Membranes

AbstractTo determine the IgGFc-binding protein (FcgammaBP) concentration in amniotic and cervical fluids in preterm prelabor rupture of membranes (PPROM) and preterm labor with intact membranes (PTL) and to assess the diagnostic indices of FcgammaBP to predict intra-amniotic infection (the presence of both microbial invasion of the amniotic cavity and intra-amniotic inflammation). In this study, we included 170 and 79 women with PPROM and PTL, respectively. Paired cervical and amniotic fluid samples were obtained using a Dacron polyester swab and transabdominal amniocentesis, respectively. The FcgammaBP concentrations in the samples were assessed using an enzyme-linked immunosorbent assay. The presence of intra-amniotic infection was associated with elevated FcgammaBP concentrations in pregnancies with PPROM and PTL [PPROM—presence: 86 ng/mL vs. absence: 13 ng/mL, p < 0.0001, area under receiver operating characteristic curve (AUC) = 0.94; PTL—presence: 140 ng/mL vs. absence: 22 ng/mL, p < 0.0001, AUC = 0.86]. In cervical fluid, the concentrations of FcgammaBP were elevated in the presence of intra-amniotic infection in pregnancies with PPROM only (presence: 345 ng/mL vs. absence: 60 ng/mL, p < 0.0001, AUC = 0.93). FcgammaBP in amniotic fluid might be a marker of intra-amniotic infection in women with both PPROM and PTL However, in cervical fluid, it is only observed in women with PPROM.

scholarly journals Identification of immunodominant epitopes on nucleocapsid and spike proteins of the SARS-CoV-2 in Iranian COVID-19 patients

2021 ◽  
Author(s):  
Faezeh Maghsood ◽  
Mohammad-Reza Shokri ◽  
Mahmoud Jeddi-Tehrani ◽  
Monireh Torabi Rahvar ◽  
Vahid Salimi ◽  
...  
Keyword(s):  
Spike Protein ◽  
Structural Proteins ◽  
Active Immunotherapy ◽  
Diagnostic Tools ◽  
Enzyme Linked Immunosorbent Assay ◽  
Native Form ◽  
Specific Antibody Response ◽  
Life Threatening ◽  
Linear Epitopes ◽  
Immunodominant Epitopes

Given the emergence of SARS-CoV-2 virus as a life-threatening pandemic, identification of immunodominant epitopes of the viral structural proteins, particularly the nucleocapsid (NP) protein and receptor binding domain (RBD) of spike protein, is important to determine targets for immunotherapy and diagnosis. In this study, epitope screening was performed using a panel of overlapping peptides spanning the entire sequences of the RBD and NP proteins of SARS-CoV-2 in the sera from 66 COVID-19 patients and 23 healthy subjects by enzyme-linked immunosorbent assay (ELISA). Also, three non-overlapping peptides belonging to the S2 domain of spike protein were assessed. Our results showed that while reactivity of patients’ sera with reduced recombinant RBD protein was significantly lower than the native form of RBD (p<0.001), no significant differences were observed for reactivity of patients’ sera with reduced and non-reduced NP protein. Pepscan analysis revealed weak to moderate reactivity towards different RBD peptide pools, which was more focused on peptides encompassing aa 181-223 of RBD. NP peptides, however, displayed strong reactivity with a single peptide covering aa 151-170. These findings were confirmed by peptide depletion experiments using both ELISA and Western blotting. Altogether, our data suggest the involvement of mostly conformational disulfide bond-dependent immunodominant epitopes in RBD-specific antibody response, while the IgG response to NP is dominated by linear epitopes. Identification of antigenic epitope in NP and RBD of SARS-CoV-2 could provide important advances for the development of passive and active immunotherapy as well as diagnostic tools for the control of COVID-19 infection.

scholarly journals Pro-cathepsin D as a diagnostic marker in differentiating malignant pleural effusion from benign pleural effusion: A retrospective cohort study

2020 ◽  
Author(s):  
Hayoung Choi ◽  
Yousang Ko ◽  
Chang Youl Lee
Keyword(s):  
Pleural Effusion ◽  
Pleural Fluid ◽  
Cathepsin D ◽  
Malignant Pleural Effusion ◽  
Operating Characteristic ◽  
Standard Procedure ◽  
Characteristic Curve ◽  
Enzyme Linked Immunosorbent Assay ◽  
Potential Biomarker ◽  
Benign Pleural Effusion

Abstract Background Malignant pleural effusion (MPE) is a common complication of lung cancer and intrathoracic spreading or metastasis of extra-thoracic malignancy. The aim of the present study was to evaluate the levels of pro-cathepsin D from plasma and pleural fluid in patients with MPE and those in patients with benign pleural effusion (BPE) including pleural tuberculosis and parapneumonic effusion.Methods This study included 81 patients with pleural effusion who underwent thoracentesis and pleural biopsy. Pleural fluid and serum were collected as a standard procedure for all individuals at the same time. The level of pro-cathepsin D was measured by the sandwich enzyme-linked immunosorbent assay method.Results Though there were no significant differences in plasma pro-cathepsin D between the two groups, the level of pleural fluid pro-cathepsin D was significantly higher in the MPE group than the BPE group (0.651 versus 0.590 pg/mL, P = 0.034) (Table 1). In addition, there were no differences in pleural fluid pro-cathepsin D level according to causative malignancy of MPE. On receiver operating characteristic curve analysis, the optimal discrimination point between the MPE group and BPE group was defined as a cut-off value of 0.5960 pg/mL for pleural fluid pro-cathepsin D (81.0% sensitivity; 53.0% specificity) and 0.4335 pg/mL for plasma pro-cathepsin D (71.4% sensitivity; 61.7% specificity).Conclusions We found that the level of pleural fluid pro-cathepsin D was significantly higher in the MPE group than the BPE group. Pro-cathepsin D could be a novel and potential biomarker to discriminate MPE from BPE.

Clinical value and potential mechanisms of LINC00221 in hepatocellular carcinoma based on integrated analysis

Epigenomics ◽  
2021 ◽  
Author(s):  
Yanlin Feng ◽  
Souraka Tapara Dramani Maman ◽  
Xinyu Zhu ◽  
Xuefang Liu ◽  
Christian Cedric Bongolo ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Operating Characteristic ◽  
Bioinformatics Analysis ◽  
Validation Cohort ◽  
Characteristic Curve ◽  
Integrated Analysis ◽  
Clinical Value ◽  
Serum Samples ◽  
Functional Roles

This study aimed to unveil the functional roles of LINC00221 in hepatocellular carcinoma (HCC). A discovery cohort and a validation cohort were respectively used to identify and verify the clinical value of LINC00221 in HCC. Bioinformatics analysis was performed to explore its potential mechanisms. LINC00221 was upregulated in HCC tissues and serum samples. Survival analysis and receiver operating characteristic curve further revealed its prognostic and diagnostic roles. Exploration of the mechanism showed that LINC00221 might exert a pro-cancer role via the lncRNA–miRNA–mRNA network. Our study reveals that upregulated LINC00221 can serve as a potential diagnostic and prognostic biomarker and provides novel clues as to the role of LINC00221 in HCC.

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