Parameters Tested for VNA validation
Documentation of the data | Data was further processed by graph pad prism version 2.10. | |
Description of the data | Data consist of several csv files. Based on each csv file a statistical analysis was done described in the publication and in metadata set to the files. File names more or less mirrow the titles within the linked publication. | |
Type of the data | Dataset | |
Total size of the dataset | 8834 | |
Author | Harzer, Maxi | |
Upload date | 2025-12-03T14:18:31Z | |
Publication date | 2025-12-03T14:18:31Z | |
Data of data creation | 2021 | |
Publication date | 2025-12-03 | |
Abstract of the dataset | Five key protocol variables were systematically assessed: (i) the applied trypsin concentration, (ii) the incubation time for proteolytic activation of the RV, (iii) the infection dose applied, (iv) the duration of neutralization time and (v) sample-specific factors that affect assay performance. Furthermore, correlation studies was done between several sample materials and Virus neutralization (VNA) and immunofourescence assay titers. Moreover the sensitivity limit and the linearity of the VNA was determined as well as intra- and inter-assay repeatability. | |
Public reference to this page | https://opara.zih.tu-dresden.de/handle/123456789/1611 | |
Public reference to this page | https://doi.org/10.25532/OPARA-906 | |
Publisher | Universität Leipzig | |
Licence | Attribution-NonCommercial-NoDerivatives 4.0 International | en |
URI of the licence text | http://creativecommons.org/licenses/by-nc-nd/4.0/ | |
Specification of the discipline(s) | 3 | |
Title of the dataset | Parameters Tested for VNA validation | |
Underlying research object | Rotaviruses | |
Software | Graph pad prism 2.10 | |
Project abstract | We optimized and standardized a virus neutralization assay (VNA) assay for RVA to evaluate its applicability across serum, intestinal mucus, and saliva samples. Five key protocol variables were systematically assessed: (i) the applied trypsin concentration, (ii) the incubation time for proteolytic activation of the RV, (iii) the infection dose applied, (iv) the duration of neutralization time and (v) sample-specific factors that affect assay performance. The following parameters have proven to be optimal: (i) end concentration of 20 µg/ml Trypsin (ii) two hours for proteolytic activation of the RV, (iii) MOI of 0,5 – 3,0 x 10-3, (iv) two or eight hours of neutralization time for serum or saliva and mucus samples. Optimization of trypsin concentration and virus activation time significantly improved assay reproducibility across sample types. Analysis of the three validated sample materials of individual swine demonstrate that RV-specific neutralizing antibodies can be reliably detected in saliva. Saliva-based neutralization titers strongly correlated with those from intestinal mucus, indicating that salivary antibodies may serve as a non-invasive surrogate for gut mucosal immunity. | |
Funding Acknowledgement | The author(s) declare financial support was received for the research and/or publication of this article. Supported by the Open Access Publishing Fund of Leipzig University. | |
Project title | Validation of a Rotavirus virus neutralization assay |
Files
Original bundle
1 - 20 of 28
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- Name:
- repeated analysis_estimation of variation_saliva.CSV
- Size:
- 98 B
- Format:
- Comma-Separated Values
- Description:
- The file contains the raw data related to the saliva-specific analyses for assessing test repeatability presented in Table 5: “Repeatability parameters of the RVA G5P[7]9-specific VNA for saliva and gut mucus” of the paper. The data table contains the number of specific result outcomes obtained from 15 repetitions of the VNA. Column 1 shows the VNT titer, and Column 2 shows the number of corresponding results. The VNA starts at a sample dilution of 2 and follows a 2-fold dilution series. For results that do not follow this 2-fold dilution series, the intra-assay results were not identical, and the mean value was calculated. The coefficient of variation was determined after linearization of the results by dividing the standard deviation by the mean.
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- Name:
- adsorbtion time mucus low titer.csv
- Size:
- 68 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 2C. The VNA was performed for each sample material in triplicate. VNA titers are shown according to the adsorption time. Kruskal-Wallis test was performed.
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- Name:
- adsorption time mucus high titer.csv
- Size:
- 76 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 2C. The VNA was performed for each sample material in triplicate. VNA titers are shown according to the adsorption time. Kruskal-Wallis test was performed.
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- Name:
- adsorption time saliva low titer.csv
- Size:
- 52 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 2C. The VNA was performed for each sample material in triplicate. VNA titers are shown according to the adsorption time. Kruskal-Wallis test was performed.
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- Name:
- adsorption time saliva medium titer.csv
- Size:
- 57 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 2C. The VNA was performed for each sample material in triplicate. VNA titers are shown according to the adsorption time. Kruskal-Wallis test was performed.
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- Name:
- adsorption time serum high titer.csv
- Size:
- 82 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 2C. The VNA was performed for each sample material in triplicate. VNA titers are shown according to the adsorption time. Kruskal-Wallis test was performed.
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- Name:
- adsorption time serum low titer.csv
- Size:
- 67 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 2C. The VNA was performed for each sample material in triplicate. VNA titers are shown according to the adsorption time. Kruskal-Wallis test was performed.
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- Name:
- Correlation within sample material RVA.csv
- Size:
- 419 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to S3: “Correlation calculations between RVA-specific VNA titers within sample materials”. VNA titer within the different sample materials of 14 pigs were determined. Spearman correlation was calculated.
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- Name:
- Correlation within sample material RVC.csv
- Size:
- 616 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 5: “Correlation calculations between RVC-specific VNA titers within sample materials”. VNA titer within the different sample materials of 15 pigs were determined. Spearman correlation was calculated.
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- Name:
- G5P[7]9 trypsin activation time.csv
- Size:
- 279 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 2B. The VNA was performed for each timepoint and concentration in triplicate. VNA titers are shown according to the activation time within a fixed trypsin concentration. Kruskal-Wallis test was performed.
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- Name:
- G5P[7]9 trypsin concentration.csv
- Size:
- 300 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 2A. The VNA was performed for each timepoint and concentration in triplicate. VNA titers are shown according to the trypsin concentration within a fixed activation time. Kruskal-Wallis test was performed.
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- Name:
- G6P[1]6 trypsin activation time.csv
- Size:
- 304 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 2B. The VNA was performed for each timepoint and concentration in triplicate. VNA titers are shown according to the activation time within a fixed trypsin concentration. Kruskal-Wallis test was performed.
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- Name:
- G6P[1]6 trypsin concentration.csv
- Size:
- 325 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 2A. The VNA was performed for each timepoint and concentration in triplicate. VNA titers are shown according to the trypsin concentration within a fixed activation time. Kruskal-Wallis test was performed.
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- Name:
- G9P[32]x trypsin activation time.csv
- Size:
- 293 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 2B. The VNA was performed for each timepoint and concentration in triplicate. VNA titers are shown according to the activation time within a fixed trypsin concentration. Kruskal-Wallis test was performed.
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- Name:
- G9P[32]x trypsin concentration.csv
- Size:
- 314 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 2A. The VNA was performed for each timepoint and concentration in triplicate. VNA titers are shown according to the trypsin concentration within a fixed activation time . Kruskal-Wallis test was performed.
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- Name:
- saliva (2) medium titer unfiltered.csv
- Size:
- 32 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 3B. The VNA titers of saliva samples with a medium titer, read out at 100% (first column) and 80% (second column), are shown. A t-test was performed.
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- Name:
- saliva (2) medium titer.csv
- Size:
- 31 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 3B. The VNA titers of saliva samples with a medium titer, read out at 100% (first column) and 80% (second column), are shown. A t-test was performed.
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- Name:
- saliva low titer 80%I.csv
- Size:
- 52 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 3B. The VNA titers of a saliva sample (read out at 80%) at different adsorption time are shown. Analysis were run in triplicates. Absolute values are shown. Standard deviation of results were calculated and t-test was performed.
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- Name:
- saliva low titer 100%I.csv
- Size:
- 52 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 3B. The VNA titers of a saliva sample (read out at 100%) at different adsorption time are shown. Analysis were run in triplicates. Absolute values are shown. Standard deviation of results were calculated and t-test was performed.
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- Name:
- saliva low titer unfiltered 80%I.csv
- Size:
- 52 B
- Format:
- Comma-Separated Values
- Description:
- This dataset corresponds to Figure 3B. The VNA titers of a saliva sample (read out at 80%) at different adsorption time are shown. Analysis were run in triplicates. Absolute values are shown. Standard deviation of results were calculated and t-test was performed.
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