scholarly journals Computationally Designed Peptides for Zika Virus Detection: An Incremental Construction Approach

Biomolecules ◽  
2019 ◽  
Vol 9 (9) ◽  
pp. 498 ◽  
Author(s):  
Marcello Mascini ◽  
Emre Dikici ◽  
Marta Robles Mañueco ◽  
Julio A. Perez-Erviti ◽  
Sapna K. Deo ◽  
...  
Keyword(s):  
Zika Virus ◽  
Dynamic Range ◽  
Acute Infection ◽  
Detection Limits ◽  
Analytical Signal ◽  
Cross Reactivity ◽  
Glycosylation Site ◽  
Protein Glycosylation ◽  
Enzyme Linked Immunosorbent Assay ◽  
Intact Virus

Herein, and in contrast to current production of anti-Zika virus antibodies, we propose a semi-combinatorial virtual strategy to select short peptides as biomimetic antibodies/binding agents for the detection of intact Zika virus (ZIKV) particles. The virtual approach was based on generating different docking cycles of tetra, penta, hexa, and heptapeptide libraries by maximizing the discrimination between the amino acid motif in the ZIKV and dengue virus (DENV) envelope protein glycosylation site. Eight peptides, two for each length (tetra, penta, hexa, and heptapeptide) were then synthesized and tested vs. intact ZIKV particles by using a direct enzyme linked immunosorbent assay (ELISA). As a reference, we employed a well-established anti-ZIKV antibody, the antibody 4G2. Three peptide-based assays had good detection limits with dynamic range starting from 105 copies/mL of intact ZIKV particles; this was one order magnitude lower than the other peptides or antibodies. These three peptides showed slight cross-reactivity against the three serotypes of DENV (DENV-1, -2, and -3) at a concentration of 106 copies/mL of intact virus particles, but the discrimination between the DENV and ZIKV was lost when the coating concentration was increased to 107 copies/mL of the virus. The sensitivity of the peptides was tested in the presence of two biological matrices, serum and urine diluted 1:10 and 1:1, respectively. The detection limits decreased about one order of magnitude for ZIKV detection in serum or urine, albeit still having for two of the three peptides tested a distinct analytical signal starting from 106 copies/mL, the concentration of ZIKV in acute infection.

scholarly journals A Rapid and International Applicable Diagnostic Device for Cobra (Genus Naja) Snakebites

Toxins ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 572
Author(s):  
Jing-Hua Lin ◽  
Wang-Chou Sung ◽  
Jiunn-Wang Liao ◽  
Dong-Zong Hung
Keyword(s):  
Detection Limits ◽  
Rapid Test ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immunochromatographic Test ◽  
Tissue Destruction ◽  
Diagnostic Device ◽  
Life Threatening ◽  
The Cross ◽  
Geographical Characteristics

Cobra snakes (genus Naja) are some of the most dangerous snake species in Asia and Africa, as their bites cause severe life-threatening respiratory failure and local tissue destruction, especially in the case of late diagnosis. The differential diagnosis of snakebite envenomation still mainly relies upon symptomatology, the patient’s description, and the experience of physicians. We have designed a rapid test, immunochromatographic test of cobra (ICT-Cobra), which obtained fair results in improving the diagnosis and treatment of Naja (N.) atra snakebites in Taiwan. In this study, we further investigated the feasibility of applying the kit for the detection of other cobra venoms based on the potential interspecies similarity. We firstly demonstrated the cross-reactivity between eight venoms of medically important cobra species and the rabbit anti-N. atra IgG that was used in ICT-Cobra by Western blotting and sandwich enzyme-linked immunosorbent assay. Then, ICT-Cobra was used to detect various concentrations of the eight venoms to elucidate its performance. Noticeable correlations between the cross-reactivity of venoms from genus Naja snakes and existing geographical characteristics were found. ICT-Cobra could detect venoms from other Asian cobras with variable detection limits comparable to those observed for N. atra, but the kit was less successful in the detection of venom from African cobras. The similar but slightly different venom components and the interaction between venom and rabbit anti-N. atra IgG led to variations in the detection limits. The transcontinental usage of ICT-Cobra might be possible due to the cross-reactivity of antibodies and similarities among the larger-sized proteins. This study showed that the close immunological relationships in the genus Naja could be used to develop a venom detection kit for the diagnosis of cobra envenomation in both Asian and African regions. Additional clinical studies and technical adjustments are still needed to improve the efficacy and broadening the application of ICT-Cobra in the future.

Development and Characterization of Anti-Estradiol Polyclonal Antibody

2012 ◽  
Vol 461 ◽  
pp. 67-70 ◽  
Author(s):  
Chao Ying Li ◽  
Jin Qing Jiang
Keyword(s):  
Polyclonal Antibody ◽  
Dynamic Range ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Uv Absorbance ◽  
Standard Curve ◽  
Ic50 Value ◽  
New Zealand White Rabbits

This paper reports an indirect competitive enzyme-linked immunosorbent assay (icELISA) using polyclonal antibody (pAb) for estradiol (E2) residues. After derivation, E2 haptens were conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) through 1-Ethyl-3-(3-dimethylaminopropy) carbodiimide (EDC) method, and New Zealand white rabbits were immunized to produce anti-E2 pAb. The conjugation ratio of E2-BSA was proved to be 18.6:1 by an UV absorbance method. Based on the square matrix titration, an icELISA standard curve was developed. The dynamic range was from 0.16 to 128 ng/mL, with LOD and IC50 value of 0.08 ng/mL and 3.76 ng/mL, respectively. Except for a little cross-reactivity (16.2%) to estrone, this assay showed negligible cross-reactivity to other analogues tested. The results suggest that the produced anti-E2 pAb could be used to develop an icELISA method for the determination of E2 residues in animal-originally products.

scholarly journals Development of Zika Virus Serological Testing Strategies in New York State

2017 ◽  
Vol 56 (3) ◽  
Author(s):  
William T. Lee ◽  
Susan J. Wong ◽  
Karen E. Kulas ◽  
Alan P. Dupuis ◽  
Anne F. Payne ◽  
...  
Keyword(s):  
Zika Virus ◽  
New York State ◽  
Denv Infection ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Testing ◽  
Recent Infection ◽  
Diagnostic Laboratory ◽  
Viral Neutralization ◽  
Testing Strategies

ABSTRACT The recent outbreak of Zika virus (ZIKV) in the Americas has challenged diagnostic laboratory testing strategies. At the Wadsworth Center, ZIKV serological testing was performed for over 10,000 specimens, using a combination of an enzyme-linked immunosorbent assay (ELISA) for IgM antibodies (Abs) to ZIKV, a polyvalent microsphere immunoassay (MIA) to detect Abs broadly reactive with flaviviruses, and a plaque reduction neutralization test (PRNT) for further testing. Overall, 42% of patients showed serological evidence of flavivirus infection (primarily past dengue virus [DENV] infection), while 7% possessed IgM Abs to ZIKV and/or DENV. ZIKV IgM Abs typically arose within 3 to 4 days, with only one instance of duration beyond 100 days after reported symptoms. PRNT analysis of 826 IgM-positive specimens showed 7% positive neutralization to ZIKV alone, 9% to DENV alone, and 85% to both ZIKV and DENV. Thus, the extensive Ab cross-reactivity among flaviviruses significantly reduced the value of performing PRNT analysis, especially when a traditional paired serum algorithm with viral neutralization titering was used. Nevertheless, the finding of a negative ZIKV result by PRNT was invaluable for reassuring both physicians and patients. The MIA detected both IgM and IgG, which enabled us to identify patients who presented without IgM anti-ZIKV Abs but still had ZIKV-specific neutralizing Abs. On the basis of these results, a new algorithm, which included an IgM Ab capture (MAC)-ELISA to detect recent infection, a flavivirus MIA to identify patients no longer producing IgM, and a single-dilution PRNT for ZIKV exclusion and occasional discrimination of ZIKV and DENV, was implemented.

scholarly journals Comparison of Four Serological Methods and Two Reverse Transcription-PCR Assays for Diagnosis and Surveillance of Zika Virus Infection

2018 ◽  
Vol 56 (3) ◽  
Author(s):  
Angel Balmaseda ◽  
José Victor Zambrana ◽  
Damaris Collado ◽  
Nadezna García ◽  
Saira Saborío ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Zika Virus ◽  
Clinical Manifestations ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Emerging Infections ◽  
Rt Pcr ◽  
Convalescent Phase ◽  
Reverse Transcription Pcr ◽  
Pcr Assays

ABSTRACTZika virus (ZIKV) is a mosquito-borne flavivirus that is responsible for recent explosive epidemics in the Americas. Notably, ZIKV infection during pregnancy has been found to cause congenital birth defects, including microcephaly, and ZIKV has been associated with Guillain-Barré syndrome in adults. Diagnosis and surveillance of Zika in the Americas have been challenging due to similar clinical manifestations and extensive antibody cross-reactivity with endemic flaviviral diseases, such as dengue. We evaluated four serological and two reverse transcription-PCR (RT-PCR) methods in acute-phase (mean day, 1.8), early-convalescent-phase (mean day, 16.7), and late-convalescent-phase (mean, ~7 months) samples from the same individuals in a long-term pediatric cohort study in Nicaragua. Well-characterized samples from 301 cases of Zika, dengue, or non-Zika, nondengue febrile illnesses were tested. Compared to a composite reference, an in-house IgM antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) and the NIAID-Biodefense and Emerging Infections (BEI) MAC-ELISA measuring IgM yielded sensitivities of 94.5% and 70.1% and specificities of 85.6% and 82.8%, respectively. The NS1 blockade-of-binding ELISA measuring anti-ZIKV NS1 antibody levels yielded sensitivities of 85.0% and 96.5% and specificities of 91.4% and 92.6% at early and late convalescence, respectively. An inhibition ELISA detecting total anti-ZIKV antibodies had sensitivity and specificity values of 68.3% and 58.3% for diagnosis and 94.0% and 98.6% for measuring annual infection incidence. Finally, the ZCD and Trioplex real-time RT-PCR assays detecting Zika, chikungunya, and dengue viruses both yielded a sensitivity of 96.1% and specificity of 100%. Together, these assays resolve the urgent need for diagnostic and surveillance tools for countries affected by Zika virus infections.

Enzyme Inhibition and Enzyme-Linked Immunosorbent Assay Methods for Carbamate Pesticide Residue Analysis in Fresh Produce

2000 ◽  
Vol 63 (12) ◽  
pp. 1758-1760 ◽  
Author(s):  
EUGENIA KATSOUDAS ◽  
HOSSNY H. ABDELMESSEH
Keyword(s):  
Enzyme Inhibition ◽  
Immunosorbent Assay ◽  
Environmental Protection Agency ◽  
Residue Analysis ◽  
Detection Limits ◽  
Cross Reactivity ◽  
Acetylcholinesterase Inhibition ◽  
Enzyme Linked Immunosorbent Assay ◽  
Food Commodities ◽  
Carbamate Pesticide

An acetylcholinesterase inhibition method was employed for detection of 21 carbamate pesticides in bananas, peaches, strawberries, and tomatoes. Each of these four agricultural commodities was spiked with 0.1 to 10 ppm of each of the 21 carbamates and individual detection levels were determined. Similar responses and detection limits were observed for all four produce when tested for a given carbamate. The detection levels ranged from 0.1 ppm for carbofuran and 3-hydroxycarbofuran to 6 ppm for promecarb and aldicarb sulfoxide. These results are generally at or below the tolerances established by the Environmental Protection Agency for these commodities. Positive samples from the enzyme inhibition screening were also analyzed with the carbaryl-specific enzyme-linked immunosorbent assay (ELISA) kit. The detection limits for carbaryl and carbofuran were 2.0 ppb and 8.0 ppb, respectively. The other carbamates did not exhibit cross-reactivity even at high ppb levels. Thus, the enzyme inhibition assay and ELISA are simple and fast screening procedures for the detection of carbamate pesticide residues in food commodities.

scholarly journals Sensitivity and Kinetics of an NS1-Based Zika Virus Enzyme-Linked Immunosorbent Assay in Zika Virus-Infected Travelers from Israel, the Czech Republic, Italy, Belgium, Germany, and Chile

2017 ◽  
Vol 55 (6) ◽  
pp. 1894-1901 ◽  
Author(s):  
Yaniv Lustig ◽  
Hana Zelena ◽  
Giulietta Venturi ◽  
Marjan Van Esbroeck ◽  
Camilla Rothe ◽  
...  
Keyword(s):  
Czech Republic ◽  
Immunosorbent Assay ◽  
Zika Virus ◽  
False Negative ◽  
Cross Reactivity ◽  
Diagnostic Methods ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibodies ◽  
Kinetics Of

ABSTRACT Serological diagnosis of Zika virus is challenging due to high cross-reactivity of Zika virus with other flavivirus antibodies. Recently, a Zika NS1-based enzyme-linked immunosorbent assay (ELISA) was developed and shown to be highly specific for Zika antibody detection; however, sensitivity was evaluated for only a small number of confirmed Zika-infected patients. In this study, we measured the sensitivity and kinetics of Zika IgM and IgG antibodies using the Zika NS1-based ELISA in 105 samples from 63 returning travelers infected with Zika virus (proven by PCR or neutralization assay) from Israel, Czech Republic, Italy, Belgium, Germany, and Chile. Zika virus IgM was detected from 2 to 42 days post-symptom onset (PSO) with an overall sensitivity of 79% in the first month and 68% until 2 months PSO, while IgG antibodies were detected from 5 days to 3 years PSO with 79% sensitivity. Interestingly, significant differences in IgM sensitivity and IgM detection period were observed between Israeli and European/Chilean Zika-infected travelers, adding to the complexity of Zika infection diagnosis and suggesting that other diagnostic methods should be complemented to reduce false-negative results.

scholarly journals Optimization and Validation of a Multiplex, Electrochemiluminescence-Based Detection Assay for the Quantitation of Immunoglobulin G Serotype-Specific Antipneumococcal Antibodies in Human Serum

2009 ◽  
Vol 16 (3) ◽  
pp. 387-396 ◽  
Author(s):  
Rocio D. Marchese ◽  
Derek Puchalski ◽  
Pamela Miller ◽  
Joseph Antonello ◽  
Olivia Hammond ◽  
...  
Keyword(s):  
Human Serum ◽  
Immunoglobulin G ◽  
Dynamic Range ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Testing Time ◽  
Igg Antibody ◽  
Meso Scale Discovery ◽  
Acceptance Criterion ◽  
Detection Assay

ABSTRACT Pneumovax 23 consists of a mixture of highly purified capsular polysaccharides (Ps) from 23 of the most prevalent serotypes of Streptococcus pneumoniae. Testing of vaccine immunogenicity has been historically performed on the enzyme-linked immunosorbent assay (ELISA) platform, validated to measure immunoglobulin G (IgG) antibodies to all 23 serotypes included in Pneumovax 23. In order to significantly improve the throughput of this form of testing, we have developed and validated a direct binding electrochemiluminescence (ECL)-based multiplex assay that can measure the antibody response in human serum to eight serotypes within a single microtiter well. The pneumococcal (Pn) ECL assay is based on the Meso Scale Discovery (MSD) technology which utilizes a Sulfo-Tag-labeled anti-human IgG antibody that emits light upon electrochemical stimulation. The Pn ECL assay exhibits a wide dynamic range and provides the ability to read concentrations down to the minimum reported concentration in the Merck ELISA (0.1 μg/ml). Cross-reactivity assessment using type-specific monoclonal antibodies showed no cross talk between antigen spots within a well. By use of the WHO Pn sample reference panel, the results obtained with the Pn ECL assay were compared to the results obtained with the international Pn ELISA. The results for the Pn ECL assay satisfied the WHO-recommended acceptance criterion for concordance for all seven serotypes with published Pn ELISA values, and the overall correlation (r value) across the seven serotypes was 0.994. The MSD methodology has great potential to be extremely useful for simultaneously quantitating IgG responses to several Pn serotypes while conserving serum volumes and laboratory testing time.

scholarly journals First detection of Zika virus infection in a Croatian traveler returning from Brazil, 2016

2017 ◽  
Vol 11 (08) ◽  
pp. 662-667
Author(s):  
Tatjana Vilibic-Cavlek ◽  
Ljiljana Betica-Radic ◽  
Giulietta Venturi ◽  
Claudia Fortuna ◽  
Stjepan Djuricic ◽  
...  
Keyword(s):  
Zika Virus ◽  
High Titer ◽  
Disease Onset ◽  
Negative Control ◽  
Cross Reactivity ◽  
Immunofluorescence Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Low Grade ◽  
Igg Antibodies ◽  
Zikv Infection

In the last few years, several imported cases of Zika virus (ZIKV) infection were reported in European countries. We report the first imported ZIKV infection case in a Croatian traveler returning from Brazil. The patient presented with a low-grade fever, pruritic rash, general weakness, myalgia, arthralgia and edema of the legs and recovered completely within a week. ZIKV infection was confirmed by detection of IgM/IgG antibodies using enzyme-linked immunosorbent assay (ELISA) and confirmed by plaque-reduction neutralization test (PRNT). ZIKV IgM antibodies cross-reacted with dengue virus (DENV), West Nile virus (WNV) and tick-borne encephalitis virus (TBEV) in ELISA. In indirect immunofluorescence assay (IFA), IgM cross-reactivity was found only with DENV-3.  ZIKV IgG antibodies cross-reacted with DENV in both ELISA and IFA. PRNT for DENV was negative. Control serology performed on days 64 and 98 after disease onset showed a decline in cross-reactive heterologous DENV IgG antibodies compared to persistently high titer of homologous ZIKV IgG antibodies.

Immunoassay of Ochratoxin A, Using Antibodies Developed Against a New Ochratoxin-Albumin Conjugate

1992 ◽  
Vol 75 (5) ◽  
pp. 824-828 ◽  
Author(s):  
Anna Breitholtz-Emanuelsson ◽  
Gunnel Dalhammar ◽  
Karl Hult
Keyword(s):  
Ochratoxin A ◽  
Immunosorbent Assay ◽  
Preparation Method ◽  
Detection Limits ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Plasma Samples ◽  
Sample Preparation Method ◽  
Ochratoxin Α ◽  
Incubation Buffer

Abstract A derivative of ochratoxin A was linked to bovine serum albumin in such a way that the carboxylic group of ochratoxin A was left unmodified. Lysine was substituted for phenylalanine in ochratoxin A, and the ε-amino group was linked to the protein. The conjugate was injected into 2 rabbits; antibodies against ochratoxin A were developed and used to develop an indirect competitive enzyme-linked immunosorbent assay. The detection limits for ochratoxin A in incubation buffer were 0.07 and 0.02 ng ochratoxin A/mL with the 2 antisera, respectively. Three hundred human plasma samples were purified by a novel sample preparation method and were analyzed by the immunosorbent assay. The detection limits for ochratoxin A in plasma samples were 0.2 and 0.1 ng ochratoxin A/mL with the 2 antisera, respectively. The cross-reactivity of the 2 antiochratoxin A sera was high for the ochratoxin A methyl ester, about 20% for ochratoxin C, and low for (4R)-4-hydroxyochratoxin A, ochratoxin α, ochratoxin B, and 4-hydroxyochratoxin B. No cross-reactivity was seen for phenylalanine and lysine.

scholarly journals Diagnosis of Zika Virus Infection by Peptide Array and Enzyme-Linked Immunosorbent Assay

mBio ◽  
2018 ◽  
Vol 9 (2) ◽  
Author(s):  
Nischay Mishra ◽  
Adrian Caciula ◽  
Adam Price ◽  
Riddhi Thakkar ◽  
James Ng ◽  
...  
Keyword(s):  
Yellow Fever ◽  
Immunosorbent Assay ◽  
Zika Virus ◽  
Natural Infection ◽  
Cross Reactivity ◽  
High Density ◽  
Serological Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Molecular Testing ◽  
West Nile

ABSTRACTZika virus (ZIKV) is implicated in fetal stillbirth, microcephaly, intracranial calcifications, and ocular anomalies following vertical transmission from infected mothers. In adults, infection may trigger autoimmune inflammatory polyneuropathy. Transmission most commonly follows the bite of infectedAedesmosquitoes but may also occur through sexual intercourse or receipt of blood products. Definitive diagnosis through detection of viral RNA is possible in serum or plasma within 10 days of disease onset, in whole blood within 3 weeks of onset, and in semen for up to 3 months. Serological diagnosis is nonetheless critical because few patients have access to molecular diagnostics during the acute phase of infection and infection may be associated with only mild or inapparent disease that does not prompt molecular testing. Serological diagnosis is confounded by cross-reactivity of immune sera with other flaviviruses endemic in the areas where ZIKV has recently emerged. Accordingly, we built a high-density microarray comprising nonredundant 12-mer peptides that tile, with one-residue overlap, the proteomes of Zika, dengue, yellow fever, West Nile, Ilheus, Oropouche, and chikungunya viruses. Serological analysis enabled discovery of a ZIKV NS2B 20-residue peptide that had high sensitivity (96.0%) and specificity (95.9%) versus natural infection with or vaccination against dengue, chikungunya, yellow fever, West Nile, tick-borne encephalitis, or Japanese encephalitis virus in a microarray assay and an enzyme-linked immunosorbent assay (ELISA) of early-convalescent-phase sera (2 to 3 weeks after onset of symptomatic infection).IMPORTANCEThe emergence of Zika virus (ZIKV) as a teratogen is a profound challenge to global public health. Molecular diagnosis of infection is straightforward during the 3-week period when patients are viremic. However, serological diagnosis thereafter of historical exposure has been confounded by cross-reactivity. Using high-density peptide arrays that tile the proteomes of a selection of flaviviruses to identify a ZIKV-specific peptide, we established two assays that enable sensitive and specific diagnosis of exposure to ZIKV. These assays may be useful in guiding clinical management of mothers at risk for potential exposure to ZIKV and enable insights into the epidemiology of ZIKV infections.

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