scholarly journals Protein Quantification, Sandwich ELISA, and Real-Time PCR Used to Monitor Industrial Cleaning Procedures for Contamination with Peanut and Celery Allergens

2004 ◽  
Vol 87 (6) ◽  
pp. 1448-1457 ◽  
Author(s):  
Oliver Stephan ◽  
Nancy Weisz ◽  
Stefan Vieths ◽  
Tanja Weiser ◽  
Burghard Rabe ◽  
...  
Keyword(s):  
Real Time ◽  
Sandwich Elisa ◽  
The United States ◽  
Protein Quantification ◽  
Enzyme Linked Immunosorbent Assay ◽  
Allergic Reactions ◽  
Cleaning Process ◽  
Cleaning Procedures ◽  
Washing Water

Abstract In the United States, peanut is one of the main sources of food allergens. Similarly, celery is a common allergenic food in Western Europe. Severe allergic reactions to both foods are common. Unexpected allergic reactions can occur after the consumption of celery-and peanut-free foods as a result of inadvertent cross-contaminations during manufacturing. Therefore, in cooperation with a flavor manufacturer, we monitored the cleaning process of slurry preparation equipment with regard to contaminations of follow-up products with celery and peanut compounds. Washing water samples taken after different cleaning steps and follow-up products were analyzed for the presence of celery and peanut traces with a celery-specific real-time polymerase chain reaction (PCR) and a peanut-specific sandwich enzyme-linked immunosorbent assay (ELISA). PCR and ELISA were compared with a nonspecific protein assay to evaluate whether the detection of protein traces can be a fast and cost-effective method for monitoring the effectiveness of wet cleaning procedures. Additionally, the allergenic potential of the celery and peanut mush, which were used as source material, were measured by a mediator release assay using a rat basophilic leukemia (RBL) cell line. In conclusion, the quantification of total protein in washing water was suitable for monitoring the cleaning process. Our study also revealed evidence that, in cases where wet cleaning is applicable, allergenic traces can be removed with high efficiency.

scholarly journals First Report of Plum pox virus (Sharka Disease) in Prunus persica in the United States

Plant Disease ◽  
2000 ◽  
Vol 84 (2) ◽  
pp. 202-202 ◽  
Author(s):  
L. Levy ◽  
V. Damsteegt ◽  
R. Welliver
Keyword(s):  
Prunus Persica ◽  
Virus Disease ◽  
Sandwich Elisa ◽  
Pcr Amplification ◽  
The United States ◽  
Enzyme Linked Immunosorbent Assay ◽  
Plum Pox Virus ◽  
Rt Pcr ◽  
Peach Fruit ◽  
First Report

Plum pox (Sharka) is the most important virus disease of Prunus in Europe and the Mediterranean region and is caused by Plum pox potyvirus (PPV). In September 1999, PPV-like symptoms were observed in peach fruit culls in a packinghouse in Pennsylvania. All symptomatic fruit originated from a single block of peach (P. persica cv. Encore) in Adams County. Trees in the block exhibited ring pattern symptoms on their leaves. A potyvirus was detected in symptomatic fruit using the Poty-Group enzyme-linked immunosorbent assay (ELISA) test from Agdia (Elkhart, IN). Reactions for symptomatic peach fruit and leaves also were positive using triple-antibody sandwich ELISA with the PPV polyclonal antibody from Bioreba (Carrboro, NC) for coating, the Poty-Group monoclonal antibody (MAb; Agdia) as the intermediate antibody, and double-antibody sandwich ELISA with PPV detection kits from Sanofi (Sanofi Diagnostics Pasteur, Marnes-La-Coquette, France) and Agdia and the REAL PPV kit (Durviz, Valencia, Spain) containing universal (5B) and strain typing (4DG5 and AL) PPV MAbs (1). PPV also was identified by immunocapture-reverse transcription-polymerase chain reaction (IC-RT-PCR) amplification and subsequent sequencing of the 220-bp 3′ noncoding region (2) (>99% sequence homology to PPV) and by IC-RT-PCR amplification of a 243-bp product in the coat protein (CP) gene (1). The virus was identified as PPV strain D based on serological typing with strainspecific MAbs and on PCR-restriction fragment length polymorphism of the CP IC-RT-PCR product with Rsa1 and Alu1 (1). This is the first report of PPV in North America. References: (1) T. Candresse et al. Phytopathology 88:198, 1998. (2) L. Levy and A. Hadidi. EPPO Bull. 24:595, 1994.

scholarly journals Development of a Sandwich Enzyme-Linked Immunosorbent Assay for Detection and Quantification of Clam Residues in Food Products

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Stef J. Koppelman ◽  
Ashley L. Lardizabal ◽  
Lynn Niemann ◽  
Joe L. Baumert ◽  
Steve L. Taylor
Keyword(s):  
Sandwich Elisa ◽  
Food Product ◽  
Food Products ◽  
Elisa Test ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Allergic Reactions ◽  
Arctica Islandica ◽  
Limit Of Quantification ◽  
Detection And Quantification

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.

scholarly journals Serological and Molecular Assays for Rapid and Sensitive Detection of Iris yellow spot virus Infection of Bulb and Seed Onion Crops

Plant Disease ◽  
2008 ◽  
Vol 92 (4) ◽  
pp. 588-594 ◽  
Author(s):  
H. R. Pappu ◽  
I. M. Rosales ◽  
K. L. Druffel
Keyword(s):  
Real Time ◽  
Rapid Detection ◽  
The United States ◽  
Iris Yellow Spot Virus ◽  
Yellow Spot ◽  
Sensitive Detection ◽  
Detection Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Spot Virus

Iris yellow spot virus (IYSV) has spread rapidly in the United States and has become an important economic constraint to the production of both bulb and seed onion crops. Symptoms caused by IYSV may be confused with those caused by other fungal and bacterial pathogens and virus-specific, reliable, sensitive, and rapid detection methods would improve the diagnosis. Antiserum was produced to Escherichia coli-expressed nucleocapsid protein of IYSV and an indirect format of the enzyme-linked immunosorbent assay (ELISA) was developed. IYSV could be detected in onion tissue at dilutions of up to 1:1,000. An IYSV-specific primer pair was designed and used in a real-time reverse-transcription polymerase chain reaction (RT-PCR) assay for the rapid detection of IYSV. Compared with standard RT-PCR, real-time RT-PCR was more rapid and sensitive. A commercially available RNA extraction kit and a total nucleic acid extraction method were compared for the quality of the templates obtained for use in real-time RT-PCR and there was no difference in limits of detection. Availability of ELISA- and PCR-based rapid and sensitive detection methods would facilitate accurate virus diagnosis and aid in better understanding of the epidemiology of the disease and in development of management strategies.

Comparison of Real-Time Polymerase Chain Reaction and Enzyme-Linked Immunosorbent Assay for Sensitive and Quantitative Detection of Hazelnuts in Nut Pastes

2018 ◽  
Vol 101 (6) ◽  
pp. 1864-1867 ◽  
Author(s):  
Ľubica Piknová ◽  
Veronika Janská ◽  
Tomáš Kuchta ◽  
Peter Siekel
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Sandwich Elisa ◽  
Pcr Analysis ◽  
Quantitative Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Wide Range ◽  
Order Of Magnitude ◽  
Polymerase Chain

Abstract Background: Hazelnuts, being a frequent agent of allergenic reactions, need to be detected in food products. Thus, it is necessary to develop and further investigate appropriate methods for detection. Objective: The aim of the study was to compare the analysis of nut pastes (peanut paste spiked with different amounts of hazelnut paste) as a model of contamination of confectionery. Methods: Real-time PCR and sandwich ELISA (RidaScreen Hazelnut Fast Kit) were used. Results: For real-time PCR, LOQ of 2 mg/kg and a quantification range from 2 to 10 000 mg/kg were determined. For ELISA, LOQ of 1 mg/kg and a quantification range from 1 to 100 mg/kg were determined. Conclusions: The comparison shows that the methods had comparable sensitivity with LOQs in the same order of magnitude. Although ELISA was slightly more sensitive, it required dilution of samples at higher concentrations of the analyte because of its narrow quantification range. Results of this study suggest that real-time PCR and ELISA are both suitable methods for the analysis of nut pastes over a wide range of concentrations. Achieved results could be useful for control as well as for technological purposes. Highlights: Real-time PCR analysis of peanut paste spiked with different amounts of hazelnut paste as a model is proposed. Sandwich ELISA analysis of peanut paste spiked with different amounts of hazelnut paste as a model is proposed. The analytical parameters of real-time PCR and ELISA methods are compared.

scholarly journals Gamma Irradiation Can Be Used To Inactivate Bacillus anthracis Spores without Compromising the Sensitivity of Diagnostic Assays

2008 ◽  
Vol 74 (14) ◽  
pp. 4427-4433 ◽  
Author(s):  
Leslie A. Dauphin ◽  
Bruce R. Newton ◽  
Max V. Rasmussen ◽  
Richard F. Meyer ◽  
Michael D. Bowen
Keyword(s):  
Gamma Irradiation ◽  
Bacillus Anthracis ◽  
Real Time ◽  
Real Time Pcr ◽  
Limit Of Detection ◽  
Sandwich Elisa ◽  
Pcr Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chromosomal Dna ◽  
Unit Reduction

ABSTRACT The use of Bacillus anthracis as a biological weapon in 2001 heightened awareness of the need for validated methods for the inactivation of B. anthracis spores. This study determined the gamma irradiation dose for inactivating virulent B. anthracis spores in suspension and its effects on real-time PCR and antigen detection assays. Strains representing eight genetic groups of B. anthracis were exposed to gamma radiation, and it was found that subjecting spores at a concentration of 107 CFU/ml to a dose of 2.5 × 106 rads resulted in a 6-log-unit reduction of spore viability. TaqMan real-time PCR analysis of untreated versus irradiated Ames strain (K1694) spores showed that treatment significantly enhanced the detection of B. anthracis chromosomal DNA targets but had no significant effect on the ability to detect targets on the pXO1 and pXO2 plasmids of B. anthracis. When analyzed by an enzyme-linked immunosorbent assay (ELISA), irradiation affected the detection of B. anthracis spores in a direct ELISA but had no effect on the limit of detection in a sandwich ELISA. The results of this study showed that gamma irradiation-inactivated spores can be tested by real-time PCR or sandwich ELISA without decreasing the sensitivity of either type of assay. Furthermore, the results suggest that clinical and public health laboratories which test specimens for B. anthracis could potentially incorporate gamma irradiation into sample processing protocols without compromising the sensitivity of the B. anthracis assays.

scholarly journals Survey of Commercial Food Products for Detection of Walnut (Juglans regia) by Two ELISA Methods and Real Time PCR

Foods ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 440
Author(s):  
Raquel Madrid ◽  
Aina García-García ◽  
Pablo Cabrera ◽  
Isabel González ◽  
Rosario Martín ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Polyclonal Antibodies ◽  
Juglans Regia ◽  
Sandwich Elisa ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Kit ◽  
Specificity And Sensitivity ◽  
Direct Elisa

Labeling of food allergens in accordance with legal regulations is important to protect the health of allergic consumers. The requirements for detecting allergens in foods involve adequate specificity and sensitivity to identify very small amounts of the target allergens in complex food matrices and processed foods. In this work, one hundred commercial samples were analyzed for walnut detection using three different methods: a sandwich enzyme-linked immunosorbent assay (ELISA) kit based on polyclonal antibodies, a direct ELISA using a recombinant multimeric scFv, and a real time PCR. The most sensitive method was real time PCR followed by sandwich ELISA kit and multimeric scFv ELISA. There was agreement between the three methods for walnut detection in commercial products, except for some heat-treated samples or those that contained pecan. The walnut ELISA kit was less affected by sample processing than was the multimeric scFv ELISA, but there was cross-reactivity with pecan, producing some false positives that must be confirmed by real time PCR. According to the results obtained, 7.0 to 12.6% of samples (depending on the analytical method) contained walnut but did not declare it, confirming there is a risk for allergic consumers. Moreover, there was one sample (3.7%) labelled as containing walnut but that tested negative for this tree nut. Genetic and immunoenzymatic techniques offer complementary approaches to develop a reliable verification for walnut allergen labeling.

scholarly journals Incidence of Brucella infection in various livestock species raised under the pastoral production system in Isiolo County, Kenya

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Josiah Njeru ◽  
Daniel Nthiwa ◽  
James Akoko ◽  
Harry Oyas ◽  
Bernard Bett
Keyword(s):  
At Risk ◽  
Longitudinal Study ◽  
Real Time ◽  
Incidence Rate ◽  
Real Time Pcr ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cox Regression Analysis ◽  
Sheep And Goats ◽  
Isiolo County

Abstract Background We implemented a longitudinal study to determine the incidence of Brucella infection in cattle, camels, sheep and goats that were being raised in a pastoral area in Isiolo County, Kenya. An initial cross-sectional survey was implemented to identify unexposed animals for follow up; that survey used 141 camels, 216 cattle, 208 sheep and 161 goats. Sera from these animals were screened for Brucella spp. using the Rose Bengal Plate test (RBPT), a modified RBPT, and an indirect multispecies Enzyme Linked Immunosorbent Assay (iELISA). Results of RBPT and iELISA were interpreted in parallel to determine seroprevalence. A total of 30 camels, 31 cattle, 22 sheep and 32 goats that were seronegative by all the above tests were recruited in a subsequent longitudinal study for follow up. These animals were followed for 12 months and tested for anti-Brucella antibodies using iELISA. Seroconversion among these animals was defined by a positive iELISA test following a negative iELISA result in the previous sampling period. All seropositive samples were further tested using real-time PCR-based assays to identify Brucella species. These analyses targeted the alkB and BMEI1162 genes for B. abortus, and B. melitensis, respectively. Data from the longitudinal study were analysed using Cox proportional hazards model that accounted for within-herds clustering of Brucella infections. Results The overall incidence rate of Brucella infection was 0.024 (95% confidence interval [CI]: 0.014–0.037) cases per animal-months at risk. Brucella infection incidence in camels, cattle, goats and sheep were 0.053 (0.022–0.104), 0.028 (0.010–0.061), 0.013 (0.003–0.036) and 0.006 (0.0002–0.034) cases per animal-month at risk, respectively. The incidence rate of Brucella infection among females and males were 0.020 (0.009–0.036) and 0.016 (0.004–0.091), respectively. Real-time PCR analyses showed that B. abortus was more prevalent than B. melitensis in the area. Results of multivariable Cox regression analysis identified species (camels and cattle) as an important predictor of Brucella spp. exposure in animals. Conclusions This study estimated an overall brucellosis incidence of 0.024 cases per animal-months at risk with camels and cattle having higher incidence than sheep and goats. These results will inform surveillance studies in the area.

67-LB: Clinical Cost Offset Analysis Comparing Real-Time CGM (RTCGM) with SMBG Based on the COMISAIR 3-Year Follow-Up Study

Diabetes ◽  
2020 ◽  
Vol 69 (Supplement 1) ◽  
pp. 67-LB
Author(s):  
JAN SOUPAL ◽  
JOHN J. ISITT ◽  
GEORGE GRUNBERGER ◽  
MARTIN PRAZNY ◽  
CHRISTOPHER PARKIN ◽  
...  
Keyword(s):  
Real Time ◽  
Follow Up Study ◽  
Cost Offset

EVALUASI KESESUAIAN PELAKSANAAN PROBITY AUDIT PADA BPKP PUSAT DENGAN PEDOMAN PROBITY AUDIT PENGADAAN BARANG/JASA PEMERINTAH

INFO ARTHA ◽  
2017 ◽  
Vol 1 (1) ◽  
pp. 17-34
Author(s):  
Fadlil Usman
Keyword(s):  
Public Sector ◽  
Real Time ◽  
Procurement Process ◽  
Assessment Activity

Probity audit is an independence assessment activity to ensure the goods/services procurement processes have been implemented consistently appropriate with the principle of upholding integrity, uprightness, honesty and fulfill certain occur legislation aimed for improving the accountability for the use of public sector fund. Probity audit is done in real time simultaneously with the goods/services procurement process. This study aims to evaluate the suitability of the implementation of probity audit conducted by BPKP Headquarter as agency that initiated the implementation of probity audit in Indonesia compared with the Probity audit Guidelines for Procurement of Goods/Services as criteria. The results of this study indicate that the implementation of probity audit conducted by BPKP Headquarter has been implemented adequately, but there are activities that do not fit the criteria, especially in the activities of the determination of the scope of the audit, the preparation of working papers and the follow-up monitoring of the audit results. Probity audit merupakan kegiatan penilaian (independen) untuk memastikan bahwa proses pengadaan barang/jasa telah dilaksanakan secara konsisten sesuai dengan prinsip penegakan integritas, kebenaran, kejujuran dan memenuhi ketentuan perundangan yang berlaku yang bertujuan meningkatkan akuntabilitas penggunaan dana sektor publik. Probity audit dilakukan secara real time yaitu bersamaan dengan pelaksanaan pengadaan barang/jasa. Penelitian ini bertujuan untuk melakukan evaluasi kesesuaian pelaksanaan probity audit yang dilakukan oleh BPKP Pusat selaku instansi yang menginisiasi pelaksanaan probity audit di Indonesia dibandingkan dengan kriteria berupa Pedoman Probity audit Pengadaan Barang/Jasa Pemerintah. Hasil dari penelitian ini menunjukkan bahwa pelaksanaan probity audit yang dilakukan oleh BPKP Pusat sudah dilaksanakan secara memadai, namun masih terdapat hal yang belum sesuai dengan kriteria terutama dalam kegiatan penentuan ruang lingkup audit, penyusunan kertas kerja dan pemantauan terhadap tindak lanjut hasil audit.

Potential Urban Rainfall Prediction Measurement System

1984 ◽  
Vol 16 (8-9) ◽  
pp. 349-362 ◽  
Author(s):  
John L Vogel
Keyword(s):  
Real Time ◽  
The United States ◽  
Monitoring Systems ◽  
Climatic Region ◽  
Convective Storm ◽  
Rainfall Prediction ◽  
Water Quality Regulations ◽  
General Data ◽  
Real Time Information ◽  
Time Information

Continued growth of urban regions and more stringent water quality regulations have resulted in an increased need for more real-time information about past, present, and future patterns and intensities of precipitation. Detailed, real-time information about precipitation can be obtained using radar and raingages for monitoring and prediction of precipitation amounts. The philosophy and the requirements for the development of real-time radar prediction-monitoring systems are described for climatic region similar to the Midwest of the united States. General data analysis and interpretation techniques associated with rainfall from convective storm systems are presented.

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