scholarly journals Comparative analysis of the allergenic characteristics and serodiagnostic potential of recombinant chitinase-like protein-5 and -12 from Sarcoptes scabiei

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Nengxing Shen ◽  
Yuhang Chen ◽  
Wenrui Wei ◽  
Lang Xiong ◽  
Yuanyuan Tao ◽  
...  
Keyword(s):  
Comparative Analysis ◽  
High Sensitivity ◽  
Specific Ige ◽  
Sarcoptes Scabiei ◽  
Positive Control ◽  
Skin Tests ◽  
Enzyme Linked Immunosorbent Assay ◽  
Allergic Reactions ◽  
Diagnostic Potential ◽  
Recombinant Chitinase

Abstract Background Scabies is caused by burrowing of the mite Sarcoptes scabiei into the stratum corneum. Currently, diagnosis via routine skin scraping is very difficult, and information on the allergenic identification of S. scabiei remains limited. Methods We performed comparative analysis of the serological diagnostic potential of recombinant S. scabiei chitinase-like protein-5 (rSsCLP5) and recombinant S. scabiei chitinase-like protein-12 (rSsCLP12) by measuring the levels of serum-specific IgG and IgE antibodies (Abs) as diagnostic markers. In addition, the allergenic characteristics of rSsCLP5 and rSsCLP12 were evaluated using IgE-binding experiments and skin tests. Results The IgE Abs-based indirect enzyme-linked immunosorbent assay (ELISA) methods showed high sensitivity and specificity: the rSsCLP5-based assay had 93.5% sensitivity and 94.4% specificity; the rSsCLP12-based assay had 100% sensitivity and 98.1% specificity. The specific IgE Abs in infested mouse sera could bind rSsCLP5 and rSsCLP12. In skin tests, rabbits in the rSsCLP5 and rSsCLP12 groups and positive control (histamine) groups exhibited allergic reactions. Most test sites in the rSsCLP12 group had edema, bleeding spots, and even ulcers or scabs, but such allergy symptoms were rare in the rSsCLP5 group. Moreover, the allergic history rabbit group had more severe allergic reactions and lower levels of IgE Abs compared to the healthy rabbit group in the same protein group. Conclusions These findings validate the use of IgE Abs to rSsCLP5 and rSsCLP12 as potentially useful markers for diagnosing scabies. Moreover, both rSsCLP5 and rSsCLP12 have allergenic properties, and the potential allergen rSsCLP12 is a stronger allergen than rSsCLP5.

scholarly journals Neo-antigens for the serological diagnosis of IgE-mediated drug allergic reactions to antibiotics cephalosporin, carbapenem and monobactam

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Edurne Peña-Mendizabal ◽  
Sergi Morais ◽  
Ángel Maquieira
Keyword(s):  
Drug Allergy ◽  
High Sensitivity ◽  
Specific Ige ◽  
In Vitro Tests ◽  
Allergic Reactions ◽  
Serum Samples ◽  
Lactam Ring ◽  
Specific Igg ◽  
Ige Mediated

Abstract New antigens deriving from -lloyl and -llanyl, major and minor determinants, respectively, were produced for β-lactam antibiotics cefuroxime, cefotaxime, ceftriaxone, meropenem and aztreonam. Twenty β-lactam antigens were produced using human serum albumin and histone H1 as carrier proteins. Antigens were tested by multiplex in vitro immunoassays and evaluated based on the detection of specific IgG and IgE in the serum samples. Both major and minor determinants were appropriate antigens for detecting specific anti-β-lactam IgG in immunised rabbit sera. In a cohort of 37 allergic patients, we observed that only the minor determinants (-llanyl antigens) were suitable for determining specific anti-β-lactam IgE antibodies with high sensitivity (< 0.01 IU/mL; 24 ng/L) and specificity (100%). These findings reveal that not only the haptenisation of β-lactam antibiotics renders improved molecular recognition events when the 4-member β-lactam ring remains unmodified, but also may contribute to develop promising minor antigens suitable for detecting specific IgE-mediated allergic reactions. This will facilitate the development of sensitive and selective multiplexed in vitro tests for drug-allergy diagnoses to antibiotics cephalosporin, carbapenem and monobactam.

scholarly journals Assessment of the IgA Immunoassay Diagnostic Potential of the Mycobacterium tuberculosis MT10.3-MPT64 Fusion Protein in Tuberculous Pleural Fluid

2010 ◽  
Vol 17 (12) ◽  
pp. 1963-1969 ◽  
Author(s):  
Leonardo Silva Araujo ◽  
Renata Maciel Moraes ◽  
Anete Trajman ◽  
Maria Helena Féres Saad
Keyword(s):  
Pleural Fluid ◽  
Histopathological Examination ◽  
False Negative ◽  
High Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Negative Results ◽  
Diagnostic Potential ◽  
False Negative Results ◽  
Sds Page ◽  
Increased Sensitivity

ABSTRACT Pleural tuberculosis (PL-TB) remains difficult to diagnose. An enzyme-linked immunosorbent assay (ELISA) was developed based on a construction containing the fusion of the Rv3019c (MT10.3) and Rv1980c (MPT64) gene sequences, and its performance was evaluated in an area where TB is endemic. A total of 92 pleural fluid (PF) samples at serial dilutions of 1:50 to 1:800 were included in the ELISA IgA MT10.3-MPT64 evaluation: 70 from TB patients and 22 from patients with other pleurisies. Confirmation of the expression and subsequent purification of the protein was made by SDS-PAGE and Western blot assays, resulting in a 36-kDa protein. ELISA IgA MT10.3-MPT64 showed sensitivities of 61.4%, 58.6%, 62.9%, 67.1%, and 70% at each PF dilution, respectively. The cumulative results of all dilutions increased sensitivity to 81.4% without jeopardizing specificity. Similar results were also obtained at the combined dilutions of 1:50, 1:200, and 1:800 or 1:50 plus 1:800 dilutions (80%). The overall sensitivity of the reference test, i.e., histopathological examination, was 74%. But, via the ELISA IgA MT10.3-MPT64 test, sensitivity was high for specimens with a negative culture (23/27; 85.2%) or nonspecific histopathology (17/18; 94.4%). Our findings demonstrated the promising use of this test as an adjunct in PL-TB diagnoses, particularly in cases with lower bacterial loads and false-negative results in the reference tests, since the new test includes such important features as quick and easy application, high sensitivity and, perhaps most importantly, affordability, which is so crucial for its widespread use in developing countries.

scholarly journals Strongyloides-Specific IgE Phage cDNA Clones and Development of a Novel ELISA for Strongyloidiasis

Diagnostics ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 985
Author(s):  
Hussain Ahmad ◽  
Norsyahida Arifin ◽  
Thomas J. Nolan ◽  
James B. Lok ◽  
Nor Suhada Anuar ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Western Blot ◽  
Rapid Test ◽  
Strongyloides Stercoralis ◽  
Specific Ige ◽  
Diagnostic Value ◽  
Epidemiological Studies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cdna Clones ◽  
Diagnostic Potential

Strongyloidiasis, caused mainly by the nematode Strongyloides stercoralis, is prevalent worldwide and potentially fatal in immunosuppressed patients. We report on a new IgE biomarker to diagnose Strongyloides infection. Sera from two groups infected with Strongyloides served as positive samples: Group 1A, in which infection was confirmed by stool-microscopy and/or stool-polymerase chain reaction (PCR) and was seropositive by an IgG-enzyme linked immunosorbent assay (ELISA) and an IgG4 rapid test, and Group 1B in which infection was confirmed by stool-PCR but was seronegative. Negative samples (controls) comprised infections with other parasites (Group II) and healthy donors (Group III). Immunoscreenings of an S. stercoralis complementary DNA (cDNA) library were performed, and the cDNA clone with the highest diagnostic potential (clone A133) was selected for recombinant protein production and then evaluated using IgE Western blot and ELISA. The Western blot showed that the recombinant protein (rA133) was 100% reactive with Group IA (n = 10) and Group IB (n = 5), and 96% non-reactive with Groups II and III (n = 25). Subsequently, the IgE-ELISA was developed and showed 100% diagnostic sensitivity in Groups IA (n = 32) and IB (n = 11); and 99.3% specificity in Groups II and III (n = 144). In conclusion, this study has identified rA133 as a novel recombinant protein with potential diagnostic value, and that the IgE-ELISA incorporating this protein may be useful for patient diagnosis and epidemiological studies.

Immediate Allergic Reactions to β-Lactams: Diagnosis and Therapy

2003 ◽  
Vol 16 (1) ◽  
pp. 19-23 ◽  
Author(s):  
A. Romano ◽  
C. Mondino ◽  
M. Viola ◽  
P. Montuschi
Keyword(s):  
Anaphylactic Shock ◽  
Specific Ige ◽  
Ring Structure ◽  
Skin Tests ◽  
In Vitro Tests ◽  
Allergic Reactions ◽  
In Vitro Test ◽  
Immunological Mechanisms

β-Lactams are the antibiotics which most frequently provoke adverse reactions mediated by specific immunological mechanisms. These reactions, classifiable as immediate or non-immediate, can be produced by the four classes of β-lactams (penicillins, cephalosporins, carbapenems and monobactams) currently available, which share a common β-lactam ring structure. Immediate reactions occur within the first hour after drug administration and are characterized by urticaria, angioedema, rhinitis, bronchospasm, and anaphylactic shock. Immediate reading skin tests are the quickest and most reliable method for demonstrating the presence of β-lactam specific IgE antibodies. It is crucial to use in diagnosis the suspected β-lactams themselves, particularly cephalosporins, in addition to penicillin determinants. Serum specific IgE assays can be used as complementary tests. Negative test results should be interpreted in light of the time elapsed from the last exposure to the responsible β-lactam. In fact, both in vivo and in vitro test sensitivity is known to decrease over time. In some diagnostic work-ups, patients with a positive history and negative skin and in vitro tests with classic reagents undergo a controlled administration of the suspected β-lactam. The management of immediate allergic reactions should take into consideration their severity and type. Adrenaline is the drug of choice in the treatment of anaphylactic shock. In addition to adrenaline, corticosteroids and antihistamines should be administered. Histamine H1 receptor antagonists are the mainstay of the treatment of immediate allergic reactions such as urticaria, rhinitis and conjunctivitis.

A novel enzyme-linked immunosorbent assay using a mixture of human native and recombinant proteinase-3 significantly improves the diagnostic potential for antineutrophil cytoplasmic antibody-associated vasculitis

2008 ◽  
Vol 68 (2) ◽  
pp. 228-233 ◽  
Author(s):  
J Damoiseaux ◽  
C Dähnrich ◽  
A Rosemann ◽  
C Probst ◽  
L Komorowski ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Odds Ratio ◽  
High Sensitivity ◽  
Antineutrophil Cytoplasmic Antibody ◽  
Antineutrophil Cytoplasmic Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Proteinase 3 ◽  
The Novel ◽  
Capture Elisa ◽  
Diagnostic Potential

Background:Antineutrophil cytoplasmic antibodies (ANCA) with a C-ANCA or P-ANCA pattern are detected in ANCA-associated vasculitis (AAV). While in most patients with AAV a C-ANCA pattern is due to reactivity with proteinase-3 (PR3)-ANCA, some C-ANCA-positive sera do not react with PR3.Objective:The development and evaluation of a direct enzyme-linked immunosorbent assay (ELISA) for PR3-ANCA with increased sensitivity.Methods:A mixture of human native (hn) and human recombinant (hr) PR3 was used as antigen coating. The resulting ELISA (anti-PR3-hn-hr) was compared with ELISAs using directly coated hn-PR3 or hr-PR3, as well as with a hn-PR3 capture ELISA. Assay characteristics were determined in patients with AAV (n = 248), with special attention for those patients with C-ANCA (n = 132), as well as disease controls (n = 585) and healthy controls (n = 429). Additionally, for prediction of relapses serial samples of 46 patients with PR3-AAV were analysed.Results:At a predefined specificity of 99% both ELISAs containing hr-PR3 revealed a substantial increase in sensitivity. For the prediction of relapses by rises in PR3-ANCA titres the capture ELISA was most optimal (odds ratio 12.5). With an odds ratio of 8.9 the novel anti-PR3-hn-hr ELISA was second best.Conclusions:Owing to the very high sensitivity of the novel anti-PR3-hn-hr ELISA for the detection of PR3-ANCA in C-ANCA-positive samples of patients with AAV this assay has an excellent diagnostic performance. This feature is combined with a good predictability of clinical relapses in patients with PR3-AAV. These characteristics challenge the dogma that, for detection of PR3-ANCA, capture ELISAs are superior for diagnosis and follow-up.

TO SEPARATELY ASSESS THE DIAGNOSTIC POTENTIAL OF TVS & MRI IN THE TERMS OF SENSITIVITY AND SPECIFICITY BY CORRELATING THE IMAGING FINDINGS WITH HISTOPATHOLOGY

2020 ◽  
Vol 4 (6) ◽  
Author(s):  
Suraj Mathur
Keyword(s):  
Transvaginal Ultrasound ◽  
High Sensitivity ◽  
Imaging Evaluation ◽  
Medical College ◽  
Conventional Mri ◽  
Diagnostic Potential ◽  
Adc Mapping ◽  
Malignant Lesions ◽  
Sensitivity Specificity

This prospective study was done in the Department of Radio diagnosis Govt. Medical College, Kozhikode. A total of 65 patients who were referred to our department with clinical suspicion of endometrial lesions and incidentally detected endometrial lesions on ultrasonography underwent transvaginal ultrasound and subsequent Imaging evaluation of pelvis MRI has very high sensitivity (95%) and specificity (98%) and is almost as accurate (97%) as histopathology in differentiating benign from malignant lesions. Addition of DWI with ADC mapping to conventional MRI increases its accuracy even more. However there is inherent limitation to MRI in detecting carcinoma in situ and micrometastasis. Keywords: TVS, MRI, Sensitivity, Specificity, Histopathology.

scholarly journals Unreliable Measurement of Basophil Maximum Leukotriene Release with the Bühlmann CAST 2000 Enzyme-Linked Immunosorbent Assay Kit

2006 ◽  
Vol 13 (3) ◽  
pp. 420-422 ◽  
Author(s):  
S. E. Burastero ◽  
C. Paolucci ◽  
D. Breda ◽  
G. Monasterolo ◽  
R. E. Rossi ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
False Positive ◽  
Positive Control ◽  
Enzyme Linked Immunosorbent Assay ◽  
Positive Results

ABSTRACT The Bühlmann CAST 2000 enzyme-linked immunosorbent assay is a potentially useful assay for measuring sulfidoleukotrienes released in vitro by allergen-challenged basophils. However, we observed that the positive-control reagent yielded positive signals in cell-free systems. These false-positive results depended on using a mouse anti-FcεRI monoclonal antibody and were prevented by degranulation-inducing reagents other than mouse monoclonal antibodies.

scholarly journals COCONUT MILK MODULATE THE ANTIGENICITY OF ALPHA-LACTALBUMIN IN BALB/C MICE

2017 ◽  
Vol 9 (3) ◽  
pp. 290 ◽  
Author(s):  
Yamina Benaissa ◽  
Samia Addou ◽  
Wafaa Dib ◽  
Omar Kheroua ◽  
Djamel Saidi
Keyword(s):  
Histological Study ◽  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Coconut Milk ◽  
Cow’S Milk ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Milk Formula ◽  
Cow's Milk

Objective: The aim of this work was to study the biochemical characteristics of coconut milk and its antigenic effect on the Balb/c mice immunized with α-lactalbumin protein, as well as its consequences on the structure of the intestinal epithelium.Methods: To achieve the objective of the study, an electrophoresis was realised on a polyacrylamide gel to determine various proteins contained in coconut milk. In addition, Lowry’s method was used to determine the amount of proteins in the formula. The antigenicity of coconut milk in sera was also studied using the Enzyme-Linked Immunosorbent Assay (ELISA) method. For the histological study, 21 w-old mice Balb/c were used and distributed in three groups of 7 mice each. Group 1, received a standard feed with no treatment (Negative control), group 2 and 3 received respectively a standard feed (Positive control) and coconut milk for a period of 28 d after being immunized with α- lactalbumin.Results: Analysis of the data revealed that the rate of proteins of cow’s milk is higher than that of the coconut milk ( p0.01). However, after carrying out the electrophoresis analysis, the coconut milk showed the absence of intact proteins. The anti α-Lactalbumin IgG titers significantly increased in positive control groups that received coconut milk (p<0.0001). Moreover, there was an increase of the intestinal villi height of mice fed with coconut milk, in the structure level of their intestinal epithelium compared to the negative control group.Conclusion: The findings of the study provide the evidence that coconut milk is a possible alternative to the cow’s milk formula in case of allergy.

Anaphylaxis to the 23-valent pneumococcal vaccine in child: a case-control study based on immediate responses in skin tests and specific IgE determination

Vaccine ◽  
2001 ◽  
Vol 19 (32) ◽  
pp. 4588-4591 ◽  
Author(s):  
Claude Ponvert ◽  
Delia Ardelean-Jaby ◽  
Anne-Marie Colin-Gorski ◽  
Bruno Soufflet ◽  
Christine Hamberger ◽  
...  
Keyword(s):  
Pneumococcal Vaccine ◽  
Case Control Study ◽  
Specific Ige ◽  
Case Control ◽  
Skin Tests ◽  
Ige Determination ◽  
Control Study

Ischemic and non-ischemic dilated cardiomyopathy

Open Medicine ◽  
2014 ◽  
Vol 9 (1) ◽  
pp. 15-20 ◽  
Author(s):  
Giovanni Fazio ◽  
Federica Vernuccio ◽  
Emanuele Grassedonio ◽  
Giuseppe Grutta ◽  
Giuseppe Lo Re ◽  
...  
Keyword(s):  
Cardiac Magnetic Resonance ◽  
Magnetic Resonance ◽  
Dilated Cardiomyopathy ◽  
High Sensitivity ◽  
Left Ventricular ◽  
Ischemic Disease ◽  
Coronary Imaging ◽  
Diagnostic Potential ◽  
Non Ischemic Dilated Cardiomyopathy ◽  
Cmr Imaging

AbstractDilated Cardiomyopathy is a high-incident disease, which diagnosis of and treatments are clinical priority. The aim of our study was to evaluate the diagnostic potential of cardiac magnetic resonance (CMR) imaging; echocardiography and the biochemical parameters that can help us differentiate between the post-ischemic and non-ischemic dilated cardiomyopathy. Materials and methods. The study enrolled 134 patients with dilated cardiomyopathy: 74 with the post-ischemic form and 60 with the non-ischemic one. All patients underwent a coronary imaging test, with echocardiogram, cardiac magnetic resonance and a blood test. Pro-inflammatory cytokines were evaluated using Luminex kit. Data was compared between the two groups. Results. Echocardiography allowed recognition of Left Ventricular Non Compaction in 2 patients. Longitudinal and circumferential strains were significantly different in the two groups (p<0.05). Using CMR imaging a post-myocarditis scar was diagnosed in 2 patients and a post-ischemic scar in 95% of patients with the chronic ischemic disease. The interleukin IL-1, IL-6 and TNF-α levels were higher in the post-ischemic group compared with the non-ischemic one. Conclusions. The use of second level techniques with a high sensitivity and specificity would help distinguish among different sub-forms of dilated cardiomyopathy.

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