Validation of MAX Aqueous Extraction on Veratox® for Total Aflatoxin ELISA Test Kit

2017 ◽  
Vol 100 (4) ◽  
pp. 1131-1133 ◽  
Author(s):  
Brooke E Roman ◽  
Dana Driksna ◽  
Mohamed M Abouzied ◽  
Frank Klein ◽  
Jennifer Rice

Abstract Neogen Corp. (Lansing, MI) has developed a common aqueous extraction method for the detection of mycotoxins in the ELISA or lateral flow format. The Veratox® for Total Aflatoxin ELISA extraction method uses a MAX 2 extraction packet and water in replacement of traditionally used organic solvents. Veratox for Total Aflatoxin has a detection range of 5–50 ppb neat or up to 300 ppb with dilution. The kit development focused on superior cross-reactivity, ability to accurately detect naturally contaminated samples, and utilization of an aqueous extraction method. In two separate validation studies, the Veratox for Total Aflatoxin test kit resulted in average yields of 91–114% in naturally contaminated mycotoxin reference material corn. The cross-reactivity profiles for aflatoxins B1, B2, G1, and G2 were 100, 113, 103, and 93%, respectively. This kit is approved by the Grain Inspection, Packers, and Stockyards Administration.

2010 ◽  
Vol 93 (2) ◽  
pp. 587-599 ◽  
Author(s):  
Anthony Lupo ◽  
Chris Roebuck ◽  
Monica Dutcher ◽  
Justina Kennedy ◽  
Mohamed Abouzied

Abstract Neogen Corp. developed the Veratox aflatoxin test kit for the detection of total aflatoxin. The purpose of this study was to validate the method under the requirements of the AOAC Research Institute Performance Tested MethodsSM (PTM) program. There are several AOAC Official MethodsSM for total aflatoxin detection in corn (994.08, 990.33, 979.18, 993.17, 990.32, 993.16, 991.31, and 990.74), varying between rapid and analytical-based methods and one rapid method that has been performance tested by the AOAC Research Institute (PTM 030701). However, the widely used reference method is AOAC Official MethodSM994.08, which is an HPLC method and is referred to as the reference method in this paper. Although considered the reference method, the HPLC procedure is complicated and requires the investment of both expensive equipment and a highly skilled technician. A rapid (e.g., ELISA) test kit to be validated by the AOAC Research Institute is needed.


Author(s):  
Brady R Cunningham ◽  
Rebecca M Coleman ◽  
Adam M Schaefer ◽  
Elizabeth I Hamelin ◽  
Rudolph C Johnson

Abstract Florida red tides have become more common and persistent in and around the Gulf of Mexico. When in bloom, red tides can produce brevetoxins in high concentrations, leading to human exposures primarily through contaminated food and ocean spray. The research described here includes adapting and validating a commercial brevetoxin water test kit for human plasma testing. Pooled plasma was fortified with a model brevetoxin, brevetoxin 3, at concentrations from 0.00500 to 3.00 ng/mL to generate calibration curves and quality control samples. The quantitative detection range was determined to be 0.0400–2.00 ng/mL brevetoxin 3 equivalents with inter- and intraday accuracies ranging from 94.0% to 109% and relative standard deviations <20%, which is within the US Food and Drug Administration guidelines for receptor-binding assays. Additionally, cross-reactivity was tested using 4 of the 10 known brevetoxins and 12 paralytic shellfish toxins. The cross-reactivity varied from 0.173% to 144% for the commercially available brevetoxin standards and 0% for the commercially available paralytic shellfish toxin standards. Fifty individual unexposed human plasma samples were measured to determine the limit of detection and endogenous interferences to the test. The validated method was used to test 31 plasma samples collected from humans potentially exposed to brevetoxins, detecting 11 positives. This method has been proven useful to measure human exposure to brevetoxins and can be applied to future exposure events.


2018 ◽  
Vol 101 (1) ◽  
pp. 102-107 ◽  
Author(s):  
Markus Lacorn ◽  
Stella Lindeke ◽  
Susanne Siebeneicher ◽  
Thomas Weiss

Abstract During the last decade, results from ELISA test kits have often been criticized as being flawed. Therefore, it may appear counterintuitive that ELISAs are used for most food allergen and gluten analytical needs. One reason, in addition to the nonavailability of comparable alternative methods, is the fact that the methods used underwent a long validation and learning period in the market. This publication presents several case studies on interference, cross-reactivity issues, calibrators, fragmented allergens and gluten, matrix influences, and misunderstood intended-use statements. Afterward, the relevant validation parameter LOD, LOQ, selectivity, and precision are discussed. Finally, a comprehensive list of practical recommendations for ELISA test kit users is presented.


Diagnostics ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 819
Author(s):  
Monique da Rocha Queiroz Lima ◽  
Raquel Curtinhas de Lima ◽  
Elzinandes Leal de Azeredo ◽  
Flavia Barreto dos Santos

In Brazil, chikungunya emerged in 2014, and by 2016, co-circulated with other arbovirosis, such as dengue and zika. ELISAs (Enzyme-Linked Immunosorbent Assays) are the most widely used approach for arboviruses diagnosis. However, some limitations include antibody cross reactivities when viruses belong to the same genus, and sensitivity variations in distinct epidemiological scenarios. As chikungunya virus (CHIKV) is an alphavirus, no serological cross reactivity with dengue virus (DENV) should be observed. Here, we evaluated a routinely used chikungunya commercial IgM (Immunoglobulin M) ELISA test (Anti-Chikungunya IgM ELISA, Euroimmun) to assess its performance in confirming chikungunya in a dengue endemic area. Samples (n = 340) representative of all four DENV serotypes, healthy individuals and controls were tested. The Anti-CHIKV IgM ELISA test had a sensitivity of 100% and a specificity of 25.3% due to the cross reactivities observed with dengue. In dengue acute cases, the chikungunya test showed an overall cross-reactivity of 31.6%, with a higher cross-reactivity with DENV-4. In dengue IgM positive cases, the assay showed a cross-reactivity of 46.7%. Serological diagnosis may be challenging and, despite the results observed here, more evaluations shall be performed. Because distinct arboviruses co-circulate in Brazil, reliable diagnostic tools are essential for disease surveillance and patient management.


Animals ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 8
Author(s):  
Marcin Śmiałek ◽  
Michał Gesek ◽  
Daria Dziewulska ◽  
Jowita Samanta Niczyporuk ◽  
Andrzej Koncicki

Transmissible viral proventriculitis (TVP) of chickens is manifested in decreased body weight gains, poor feed conversion and weight diversity. Although TVP etiology has not been defined, a Birnaviridae family member, named chicken proventricular necrosis virus (CPNV) is considered as a potential factor of a disease. This study was undertaken in order to reproduce TVP and to evaluate its etiology. Broiler chickens of the TVP-infected group were inoculated with TVP positive proventriculi homogenate on the 24th day of life. Samples were collected, on infection day and 14 days post-infection (dpi). The 14 dpi anatomo- and histopathological evaluation, revealed that we have succeeded to reproduce TVP. TVP-infected birds gained 30.38% less body weight. In the TVP-infected group a seroconversion against picornaviruses, fowl adenoviruses (FAdV) and infectious bursal disease viruses (IBDV) was recorded with an ELISA test. Using RT-PCR and PCR, CPNV was detected in proventriculi and FAdV in spleens and livers of infected birds, 14 dpi. Our study supports that CPNV is involved in the development of TVP. We did not record the presence of IBDV in TVP or control birds, despite our recording of a seroconversion against IBDV in TVP infected birds. CPNV and IBDV belong to the same family, which allows us to assume serological cross-reactivity between them. The role of FAdV needs further evaluation.


2014 ◽  
Vol 955-959 ◽  
pp. 3377-3380 ◽  
Author(s):  
Qing Chen ◽  
Te Wang ◽  
Jin Dan Zhao ◽  
Fu Hui Kang ◽  
Yi Ming Zhang ◽  
...  

The simultaneously co-production of Poly-β-hydroxybutyrate (PHB) and ectoine in a process (PHB/Ect co-production) and co-products extraction have great significant for reducing the manufacture cost and promoting industrialization of PHB and ectoine. The pure aqueous extraction method based on osmotic downshock was used for the extraction of PHB/Ect co-products byH. salina. The effects of osmotic pressure, extraction temperature and extraction time on the extraction rate of PHB were investigated. The ectoine was extracted and purified by the techniques of hollow fiber and ion exchange. The optimal conditions for PHB extraction were osmotic downshock in pure water, extraction temperature at 60 °C and extraction for 4 h. The extraction rate of PHB was 87.5%. The extraction rate of ectoine was 84.2%.


2013 ◽  
Vol 3 (5) ◽  
pp. 390-393 ◽  
Author(s):  
SR Kandel ◽  
P Ghimire ◽  
BR Tiwari ◽  
M Rajkarnikar

Background: HIV and Hepatitis B infections are public health problems in Nepal. This study was conducted based at NRCS/CBTS, with the objective of determining the HIV and HBsAg sero-prevalence in non-remunerated volunteer blood donors. Materials and Methods: A total of 66,904 units of blood collected, following donor recruitment criteriaduring March 2009-Sept. 2010 was included for analysis. All donated blood samples were subjected to screening for Transfusion transmitted infections including HIV and Hepatitis B surface antigen using standard ELISA test kits (Dade Behring, Germany). Initial reactive sera were re-tested for reconfi rmation with same test kits plus another test kit (Detect-HIV, Adaltis Inc, and Qualisa). Results: Out of 66,904 units of blood collected, 56,973 units were from male and 9,931 were from female donors. Among the total screened samples, 73 (0.10%) were found to be positive for HIV, {0.11% (64/56973) in male and 0.09% (9/9931) in female}; the difference between male and female donors (?2<3.841) was statistically signifi cant. The seroprevalence of HIV was highest in age group of 30- 39 both in male and female (p<0.001). Similarly, for HBsAg, overall seroprevalence was found to be 0.47% (316/66904 {0.42% (242/56973) in male and 0.74% (74/9931) in female}. The difference was statistically signifi cant (?2<3.841). The highest HBsAg sero-prevalence(0.65%) was also observed in same age group i.e. 30-39 (p<0.001) in male but highest seroprevalence (2.63%) was observed inage group of ?50 in female. Conclusion: Both HIV and HBV sero-prevalence is high in adult voluntary blood donors. Journal of Pathology of Nepal (2013) Vol. 3, No.1, Issue 5, 390-393 DOI: http://dx.doi.org/10.3126/jpn.v3i5.7864


Author(s):  
Omid Nekouei ◽  
Paolo Martelli ◽  
Sophie St-Hilaire ◽  
Hui Suk Wai ◽  
Karthiyani Krishnasamy ◽  
...  

Rabies is a fatal zoonotic disease that can affect all mammals. Following the directives of the rabies ordinance of the Government of Hong Kong, all wild macaques captured under an ongoing sterilization program (since 2000) were vaccinated against rabies. The main objective of this study was to assess the serological response to rabies vaccination in the population of Hong Kong macaques. An inactivated rabies vaccine was subcutaneously administered to captured macaques under anesthesia. In a 2015 field survey, blood samples from the animals were collected and stored in -80℃ freezer. In July 2021, all frozen sera from vaccinated animals were prepared and tested for antibodies against rabies virus using a commercial enzyme-linked immunosorbent assay (ELISA) test. The test results were dichotomized at the recommended cut-off point of the test kit. Sixty-five samples from the vaccinated macaques were available for this study. All of these animals had received at least one dose of vaccine (1 vaccination) between 2008 and 2015. The interval between the 1 vaccination and blood sampling dates ranged from 21 to 2,779 days. Only five of the 65 macaques had a second vaccination record at the time of sampling; all five had high antibody levels. Among the remaining macaques, 77% (46/60) were positive for rabies antibodies. No specific association was observed between the post-vaccination period and the antibody titer of these macaques and no adverse reactions to vaccination were reported. The current vaccination strategy in Hong Kong macaques appears to effectively elicit rabies antibodies in a high proportion of macaque populations in the wild (78-87%). However, reaching the precise level of protection against a potential challenge with the virus should further be investigated.


Author(s):  
Sabina A. Seidova

In paper the results of the carried out analysis of literary data on preparation of motor fuels of high quality by extraction method of purification of the corresponding oil distillates with use of compounds of various class as a selective solvent have been presented. In particular, the results of comparative analysis of existing methods of the extraction purification of distillates of motor fuels from unnecessary components – aromatic hydrocarbons, sulphur-containing compounds and resinous substances with use of organic solvents and ion-liquid compositions as a selective solvent have been presented. The advantage of the extraction method of purification of motor fuels determined by possibility of the process at low temperature and pressure, by absence of necessity of application of the expensive catalysts, by possibility of regeneration and reuse of solvent, etc. in comparison with widely used hydrogenation method has been shown. The lacks of the organic solvents used as an extractant have been also listed and due to the ecological problems the use of non-volatile, thermally stable ion-liquid compositions as a selective solvent in the processes of purification of the distillates, intended for preparation a high quality target products, such as diesel fuel, gasoline, base oils for various purposes has been substantiated. In paper the results of systematic investigations carried out at the Institute of Petrochemical Processes of Azerbaijan National Academy of Sciences with the participation of the authors themselves, concerning the selective purification of the oil fractions of various composition and viscosity with use of ionic liquids synthesized on the basis of formic and acetic acids composition have been also presented. By carried out analysis it has been shown the perspectivity of application of the ion-liquid compositions as an extractant in the processes of the selective purification of the oil distillates.


2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Stef J. Koppelman ◽  
Ashley L. Lardizabal ◽  
Lynn Niemann ◽  
Joe L. Baumert ◽  
Steve L. Taylor

Seafood is a frequent cause of allergic reactions to food globally. The presence of undeclared trace amounts of clam can cause allergic reactions in sensitive individuals. Limited tools are available to test food products for the presence of traces of clam. We report on the development of a sandwich ELISA that can detect and quantify clam protein in food. Antisera against a mix of two commercially important clam species, Atlantic Surf (Spisula solidissima) and ocean quahog (Arctica islandica), were raised in rabbit and sheep. A sandwich ELISA was constructed with this antisera, and sensitivity and specificity were evaluated. Also, model food products spiked with clam protein were analyzed to assess the performance of the ELISA. Comparison was made with a commercially available ELISA for crustacea. The lower limit of quantification of the sandwich ELISA is 2.5 ppm clam protein in food samples, allowing the detection of low amounts of clam that may trigger a reaction in clam allergic patients. The sandwich ELISA was highly specific with cross-reactivity only noted for other molluscan shellfish (mussel and scallop). Clam protein in tomato juice and potato cream soup was detected well with recoveries ranging from 65 to 74% and from 74 to 113%, respectively. However when potato cream soup was retorted, the recover fell to 20%, imposing the risk of underestimating the clam content of a food product. A commercially available crustacean ELISA test was not suitable to detect clam protein. The sandwich ELISA described here is suitable for detection and quantification of clam protein in food products. Care should be taken with food products that have been retorted as the results may be underestimated.


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