scholarly journals Designing a Recombinant Multi-Epitope Antigen of Echinococ-cus granulosus to Diagnose Human Cystic Echinococcosis

2020 ◽  
Author(s):  
Aliyar MIRZAPOUR ◽  
Seyyed Javad SEYYED TABAEI ◽  
Mojgan BANDEHPOUR ◽  
Ali HAGHIGHI ◽  
Bahram KAZEMI
Keyword(s):  
Sensitivity And Specificity ◽  
Cystic Echinococcosis ◽  
Severe Disease ◽  
Epitope Prediction ◽  
Elisa Test ◽  
Serum Samples ◽  
Sds Page ◽  
Immunodiagnostic Tests ◽  
Human Cystic Echinococcosis ◽  
Commercial Kit

Background: Cystic echinococcosis can cause severe disease and probable death in humans. Epitopes of its antigens play a key role in the sensitivity and specificity of immunodiagnostic tests. Methods: Epitope prediction software programs predict the most antigenic linear B-cell epitopes of AgB (8 kD), Ag5, and Ag95. Six such epitopes were predicted and connected by “Gly-Ser” linker and synthesized. The purity of the concentrated recombinant multi-epitope protein was assessed by 15% SDS-PAGE. Overall, 186 serum samples were collected from the Loghman Hakim Hospital and different laboratories, Tehran, Iran, from July 2016 to February 2017. Patients infected with hepatic hydatid cysts, patients infected by other parasites and viruses, and healthy individuals were used to detect the anti-CE IgG using recombinant multi-epitope protein. Results: Forty-one samples out of 43 cases of hydatidosis were diagnosed correctly as positive, and two were negative. In addition, six negative cases of healthy individual group were diagnosed as positive and negative with rMEP-ELISA and the commercial kit, respectively. Therefore, these six samples were considered as false positive using our method. In addition, a diagnostic sensitivity of 95.3% (95% CI, 84.19% to 99.43%) and a specificity of 95.0% (95% CI, 89.43% to 98.14%) were obtained using optimum cutoff value (0.20). The sensitivity and specificity of the commercial kit was 100%. Conclusion: Our findings showed high diagnostic accuracy of the ELISA test using the developed recombinant protein, which encourages the use of this recombinant multi-epitope protein for rapid serological diagnosis of hydatidosis.

Seroprevalence of Human Cystic Echinococcosis in Sanandaj City, Kurdistan Province, Western Iran

2021 ◽  
Author(s):  
Naser Nazari ◽  
Tooran Nayeri ◽  
Farkhondeh Hazrati
Keyword(s):  
Sex Education ◽  
Cystic Echinococcosis ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cluster Sampling ◽  
Serum Samples ◽  
Cystic Hydatid Disease ◽  
Western Iran ◽  
Significant Difference ◽  
Human Cystic Echinococcosis

Background and Aims: Echinococcus granulosus (E. granulosus) is a cestode parasite that causes cystic hydatid disease in humans worldwide. Iran is one of the endemic regions for infection that indicate the importance and presence of infection in this country. Therefore, the current research aimed to characterize the seroprevalence of human cystic echinococcosis in Sanandaj city, Kurdistan province, western Iran. Materials and methods: Totally, 500 serum samples were collected from patients referred to different health centers in Sanandaj city using cluster sampling in 2018-2019. All the sera were examined using the enzyme-linked immunosorbent assay test. Results: The seroprevalence of human hydatidosis was reported at 2.2% by ELISA test in Sanandaj city. This rate was 9 (1.9%) in women and 2 (0.4) in men. The age group of 20-30 years old had the highest positivity rate (1.0%). Also, the subjects that consumed home slaughtered meat had the highest infection rate at 4 (0.8%). There was no significant difference regarding factors studied such as sex, education, residence, consumed water, keeping a dog, and the seropositivity. Conclusions: Seroprevalence of human cystic echinococcosis in Sanandaj city is lower than the general prevalence in Iran. Our research team hopes to provide accurate data on the prevalence of hydatidosis in Sanandaj encourage more extensive research to help prevent this parasite in Iran and worldwide.

scholarly journals Analytical and diagnostic performances of a high-throughput immunoassay for SARS-CoV-2 IgM and IgG

2020 ◽  
Author(s):  
Andrea Padoan ◽  
Chiara Cosma ◽  
Paolo Zaupa ◽  
Mario Plebani
Keyword(s):  
Sensitivity And Specificity ◽  
Symptom Onset ◽  
Severe Disease ◽  
Onset Time ◽  
Time Frame ◽  
Serum Samples ◽  
Time Period ◽  
Analytical Imprecision ◽  
Range Of Values

BackgroundAbstractReliable SARS-CoV-2 serological assays are required for diagnosing infections, for the serosurveillance of past exposures and for assessing the response to future vaccines. In this study, the analytical and clinical performances of a chemiluminescent immunoassays for SARS-CoV-2 IgM and IgG detection (Mindray CL-1200i), targeting Nucleocapsid (N) and receptor binding domain (RBD) portion of the Spike protein, were evaluated.MethodsPrecision and linearity were evaluated using standardized procedures. A total of 157 leftover serum samples from 81 hospitalized confirmed COVID-19 patients (38 with moderate and 43 with severe disease) and 76 SARS-CoV-2 negative subjects (44 healthcare workers, 20 individuals with rheumatic disorders, 12 pregnant women) were included in the study. In an additional series of 44 SARS-CoV-2 positive, IgM and IgG time kinetics were also evaluated in a time-period of 38 days.ResultsPrecision was below or equal to 4% for both IgM and IgG, in all the studied levels, whilst a slightly significant deviation from linearity was observed for both assays in the range of values covering the manufacturer’s cut-off. Considering a time frame ≥ 12 days post symptom onset, sensitivity and specificity for IgM were 92.3% (95%CI:79.1%-98.4%) and 92.1% (95%CI:83.6%-97.0%). In the same time frame, sensitivity and specificity for IgG were 100% (95%CI:91.0%-100%) and 93.4% (95%CI:85.3%-97.8%). The assays agreement was 73.9% (Cohen’s kappa of 0.373). Time kinetics showed a substantial overlapping of IgM and IgG response, the latter values being elevated up to 38 days from symptoms onset.ConclusionsAnalytical imprecision is satisfactory as well as the linearity, particularly when taking into account the fact that both assays are claimed to be qualitative. Diagnostic sensitivity of IgG was excellent, especially considering specimens collected ≥12 days post symptom onset. Time kinetics suggest that IgM and IgG are detectable early in the course of infection, but the role of SARS-CoV-2 antibodies in clinical practice still requires further evaluations.

Standardization and characterization of antigens for the diagnosis of aspergillosis

2012 ◽  
Vol 58 (4) ◽  
pp. 455-462 ◽  
Author(s):  
Cheila Denise Ottonelli Stopiglia ◽  
Alicia Arechavala ◽  
Mariana Carissimi ◽  
Julia Medeiros Sorrentino ◽  
Valério Rodrigues Aquino ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Aspergillus Niger ◽  
Sensitivity And Specificity ◽  
Predictive Value ◽  
High Sensitivity ◽  
Antigenic Activity ◽  
Sds Page ◽  
Mascot Search Engine ◽  
Commercial Kit

The aim of this study was to develop and characterize antigens for the diagnosis of aspergillosis. Nine strains of Aspergillus species Aspergillus fumigatus , Aspergillus flavus , and Aspergillus niger were grown in Sabouraud and Smith broth to produce exoantigens. The antigens were tested by immunodiffusion against sera from patients with aspergillosis and other systemic mycoses. The protein fraction of the antigens was detected by SDS–PAGE; Western blot and representative bands were assessed by mass spectrometry coupled to a nano Acquity UltraPerformance LC and analyzed by the Mascot search engine. Concurrently, all sera were tested with Platelia Aspergillus EIA. The most reactive antigens to sera from patients infected by A. fumigatus were produced by A. fumigatus MG2 Sabouraud and pooled A. fumigatus Sabouraud samples, both with a sensitivity of 93% and specificity of 100% and 97%, respectively. Aspergillus niger and A. flavus antigens were reactive against A. niger and A. flavus sera, each one with a sensitivity and specificity of 100%. Two proteins, probably responsible for antigenic activity, β-glucosidase in A. fumigatus and α-amylase in A. niger were attained. The commercial kit had a specificity of 22%, sensitivity of 100%, positive predictive value of 48%, and negative predictive value of 100%. The antigens produced showed high sensitivity and specificity and can be exploited for diagnostics of aspergilloma.

scholarly journals Metabolomics and Inflammatory Mediator Profiling for the Differentiation of Life-Threatening and Non-Severe Appendicitis in the Pediatric Population

Metabolites ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 664
Author(s):  
Nusrat Shommu ◽  
Jaime Blackwood ◽  
Craig Jenne ◽  
Ari Joffe ◽  
Dori-Ann Martin ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Pediatric Population ◽  
Severe Disease ◽  
Pediatric Intensive Care ◽  
Proton Nuclear Magnetic Resonance ◽  
Gas Chromatography Mass Spectrometry ◽  
Diagnostic Tools ◽  
Serum Samples ◽  
Inflammatory Protein ◽  
Life Threatening

Background: While children with appendicitis often have excellent clinical outcomes, some develop life-threatening complications including sepsis and organ dysfunction requiring pediatric intensive care unit (PICU) support. Our study applied a metabolomics and inflammatory protein mediator (IPM) profiling approach to determine the bio-profiles of children who developed severe appendicitis compared with those that did not. Methods: We performed a prospective case-control study of children aged 0–17 years with a diagnosis of appendicitis. Cases had severe disease resulting in PICU admission. Primary controls had moderate appendicitis (perforation without PICU); secondary controls had mild appendicitis (non-perforated). Serum samples were analyzed using Proton Nuclear Magnetic Resonance (1H NMR) Spectroscopy and Gas Chromatography-Mass Spectrometry (GC-MS); IPM analysis was performed using plasma bead-based multiplex profiling. Comparisons were made using multivariate data statistical analysis. Results: Fifty-three children were included (15 severe, 38 non-severe). Separation between severe and moderate appendicitis demonstrated excellent sensitivity and specificity (100%, 88%; 14 compounds), separation between severe and mild appendicitis also showed excellent sensitivity and specificity (91%, 90%; 16 compounds). Conclusions: Biomarker patterns derived from metabolomics and IPM profiling are capable of distinguishing children with severe appendicitis from those with less severe disease. These findings provide an important first step towards developing non-invasive diagnostic tools for clinicians in early identification of children who are at a high risk of developing severe appendicitis.

scholarly journals Rickettsial neglected zoonoses: prevalence of scrub typhus at central Karnataka

2017 ◽  
Vol 5 (8) ◽  
pp. 3672
Author(s):  
Vinod Kumar C. S. ◽  
Satish Patil ◽  
B. S. Prasad ◽  
N. K. Kalappanavar ◽  
S. G. Jayaraj ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Fever Of Unknown Origin ◽  
Scrub Typhus ◽  
Unknown Origin ◽  
Blood Parameters ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Igm Antibodies ◽  
Igm Elisa

Background: Fever of unknown Origin (FUO) has many multiple causes such as enteric fever, malaria, dengue, tuberculosis, brucellosis. But scrub typhus is less known cause in Indian scenario. The present study reports the prevalence of scrub typhus at central Karnataka and compares the sensitivity and specificity of Weil-Felix test and the IgM ELISA in the detection of infection.Methods: 368 serum samples of FUO cases were collected. Weil-Felix test was performed and also analyzed for IgM antibodies to Orienta tsutsugamushi by IgM ELISA test along with haematological and biochemical investigations.Results: Out of 368 patients of fever of unknown origin, 94 cases were positive by OXK antigens by Weil Felix test and 61 were positive by ELISA test for ST IgM antibodies. Fever was the most common clinical presentation occurring in ST IgM ELISA positive cases, followed by myalgia in 90.1% cases, headache in 77%, hepatomegaly in 65.5%, splenomegaly in 62.2% and rashes were seen in 29.5% patients. Eschar was seen in 13.1% patients, pneumonia in 3.2% and meningo-encephalitis in 1.6%. Sensitivity and specificity of WFT in relation to IgM ELISA at a titre of 160 was 81.97% and 85.67% respectively.Conclusions: With the growing number of cases detected in India, scrub typhus is fast emerging as a public health threat and also due to limited diagnostics leading to underreporting, Weil Felix test could be used in adjunct with Enzyme-linked immunosorbent assay and blood parameters in the diagnosis of rickettsial diseases.

Development of 2 types of competitive enzyme-linked immunosorbent assay for detecting antibodies to the rinderpest virus using a monoclonal antibody for a specific region of the hemagglutinin protein

2007 ◽  
Vol 53 (6) ◽  
pp. 720-726 ◽  
Author(s):  
S. Khamehchian ◽  
R. Madani ◽  
M.J. Rasaee ◽  
F. Golchinfar ◽  
R. Kargar
Keyword(s):  
Monoclonal Antibody ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Specific Region ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rinderpest Virus ◽  
Serum Samples ◽  
Positive Serum ◽  
H Protein ◽  
Commercial Kit

A competitive enzyme-linked immunosorbent assay (C-ELISA) has been developed and standardized for the detection of antibodies to the rinderpest virus (RPV) in sera from cattle, sheep, and goats. The test is specific for rinderpest because it does not detect antibodies to peste-des-petits-ruminants virus (PPRV). The test depends on the ability of the monoclonal antibody (MAb) directed against the hemagglutinin (H) protein of RPV to compete with the binding of RPV antibodies in the positive serum to the H protein of this virus. This MAb recognized a region from amino acids 575 to 583 on the H protein of RPV that is unique to the RPV H protein and is not present on the hemagglutinin-neuraminidase protein of PPRV. Another C-ELISA (peptide C-ELISA) was set up using this specific region as an antigen. A threshold value of 64.4% inhibition was established for the RPV C-ELISA, with 90 known RPV-negative and 30 RPV-positive serum samples. Using common serum samples, a cutoff value of 43.0% inhibition for the peptide C-ELISA was established. Based on statistical analysis, the overall sensitivity and specificity of the RPV C-ELISA, relative to those of a commercial kit, were found to be 90.00% and 103.33%, respectively. However, the sensitivity and specificity of the peptide C-ELISA were found to be 180.00% and 73.33%, respectively. Although a common MAb in 2 new C-ELISA systems was used, variation in their percent inhibition, due to the use of different antigens, was observed. Taking into consideration the difference in percent inhibition of the 2 described assays and the commercial kit (50%), it was found that the RPV C-ELISA and the peptide C-ELISA are more specific and sensitive tools than the commercial kit for assessing herd immune status and for epidemiologic surveillance.

MONITORING STUDIES OF ARBOVIRUS INFECTIONS TRANSMITTED BY MOSQUITOES ON THE TERRITORY OF THE VOLGOGRAD REGION

2019 ◽  
pp. 60-66
Author(s):  
E.V. Molchanova ◽  
D.N. Luchinin ◽  
A.O. Negodenko ◽  
D.R. Prilepskaya ◽  
N.V. Boroday ◽  
...  
Keyword(s):  
Virus Antigen ◽  
Elisa Test ◽  
Serum Samples ◽  
Blood Sucking ◽  
Tick Borne Encephalitis ◽  
Volgograd Region ◽  
Two Samples ◽  
Probable Presence ◽  
California Serogroup ◽  
Tick Borne Encephalitis Virus

The paper presents data from the monitoring studies’ results of arbovirus infections transmitted by mosquitoes in the Volgograd region. West Nile virus antigen (WNV) in 9 samples, Tahyna virus in one sample, Batai virus in two samples were detected in the study of 110 samples of field material (blood-sucking mosquitoes) by ELISA test. Antibodies to WNV in 16.58 percent of the samples, to tick-borne encephalitis virus in 1.08 percent, to viruses of the California serogroup and Ukuniemi in 1.09 percent, to the virus Sindbis in 2.17 percent were detected as a result of the study of blood serum samples from donors in the Volgograd region. Thus, we obtained data on the probable presence of the Batai, Sindbis, Ukuniemi and Californian serogroup viruses along with the circulation of WNV on the territory of the Volgograd region.

scholarly journals Human Cystic Echinococcosis in Lebanon: A Retrospective Study and Molecular Epidemiology

2021 ◽  
Author(s):  
Gaelle Joanny ◽  
Maria Grazia Cappai ◽  
Francesca Nonnis ◽  
Claudia Tamponi ◽  
Giorgia Dessì ◽  
...  
Keyword(s):  
Echinococcus Granulosus ◽  
Cystic Echinococcosis ◽  
Epidemiological Data ◽  
Sensu Stricto ◽  
Control Measures ◽  
Mediterranean Countries ◽  
High Gene ◽  
Human Infections ◽  
And Control ◽  
Human Cystic Echinococcosis

Abstract Purpose Human cystic echinococcosis (CE) is a zoonotic parasitic disease that constitutes a public health challenge and a socio-economic burden in endemic areas worldwide. No specific surveillance system of CE infections in humans exists in Lebanon. The incidence and trends over time have not been documented. The current study aimed to assess the demographic and epidemiologic features of human CE surgical cases over a 14-year period in the five main regions of Lebanon. Methods From 2005 to 2018, a total of 894 surgically confirmed cases of hydatidosis were recorded from five anatomy and pathology laboratories. Results The mean annual surgical incidence was 1.23/100,000 inhabitants. Over the span of these years, the incidence increased from 0.53 to 1.94 cases/100,000 inhabitants in 2005 and 2018, respectively. CE is present in Lebanon with an uneven distribution from one region to the other with higher prevalence in Bekaa (29.0%), a rural area where sheep raising is widespread. Human CE cases were more common in females (60.1%) than in males (39.9%) and a high burden of infection was reported for the age group of 30–39 years. Besides, 66.7% of the cases expressed only liver complications whereas, 20.5% showed predilection towards lungs. The 7.8% of cases presented cysts in other organs, and 1.3% showed multiple localizations. Additionally, predominant involvement of Echinococcus granulosus sensu stricto was recorded in human infections. Comparison of Echinococcus granulosus s.s. populations from different Mediterranean countries also revealed high gene flow among this region and sharing of alleles. Conclusion The current study is a step forward to fill the gap of knowledge for the hydatidosis in Lebanon where the lack of epidemiological data and control measures have resulted in higher incidence of human CE. Graphic Abstract

Characterization of excretory–secretory products from protoscoleces of Echinococcus granulosus and evaluation of their potential for immunodiagnosis of human cystic echinococcosis

Parasitology ◽  
2004 ◽  
Vol 129 (3) ◽  
pp. 371-378 ◽  
Author(s):  
D. CARMENA ◽  
J. MARTÍNEZ ◽  
A. BENITO ◽  
J. A. GUISANTES
Keyword(s):  
Echinococcus Granulosus ◽  
Cystic Echinococcosis ◽  
Secretory Protein ◽  
Major Protein ◽  
Healthy Donors ◽  
Secretory Products ◽  
Protein Components ◽  
Human Cystic Echinococcosis

This study describes, for the first time, the characterization of excretory–secretory antigens (ES-Ag) from Echinococcus granulosus protoscoleces, evaluating their usefulness in the immunodiagnosis of human cystic echinococcosis. ES-Ag were obtained from the first 50 h maintenance of protoscoleces in vitro. This preparation contained over 20 major protein components which could be distinguished by 1-dimensional SDS–PAGE with apparent masses between 9 and 300 kDa. The culture of of protoscoleces from liver produced a greater variety of excretory–secretory protein components than those from lung. Determination of enzymatic activities of secreted proteins revealed the presence of phosphatases, lipases and glucosidases, but no proteases. These findings were compared to those obtained from somatic extracts of protoscoleces and hydatid cyst fluid products. Immunochemical characterization was performed by immunoblotting with sera from individuals infected by cystic echinococcosis (n=15), non-hydatidic parasitoses (n=19), various liver diseases (n=24), lung neoplasia (n=16), and healthy donors (n=18). Antigens with apparent masses of 89, 74, 47/50, 32, and 20 kDa showed specificity for immunodiagnosis of human hydatidosis. The 89 and 74 kDa components corresponded to antigens not yet described in E. granulosus, whereas proteins of 41–43 kDa and 91–95 kDa were recognized by the majority of the non-hydatid sera studied.

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