Optimizing Release of Nucleic Acids of African Swine Fever Virus and Influenza A Virus from FTA Cards

Figures

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  Figure 1. (A) Comparison of 11 different releasing methods for the ASFV genome detection with and without nucleic acid extraction. The mean Ct values based on the PCR results of all four dilution steps tested with seven FTA cards are shown. The used methods are described in the legend of Table 1. SD analysis was carried out (number of replicates = 11, Supplementary Table S1) are shown. The standard deviation (SD) value for all methods by the direct qPCR amplification is 1.96 and for qPCR amplification with extraction is 1.72. The standard error of the mean value for all methods by direct amplification is 0.59 and for the amplification with extraction is 0.51. An unpaired multiple t-test was performed to test the significance between the different RNA releasing methods based on the both qPCR amplification direct and with extraction with a resulting p-value > 0.99 for the taken values, which is not statistically significant. (B) Comparison of 11 different releasing methods for the IAV genome detection with and without nucleic acid extraction. The mean Ct values based on RNA-releasing methods of all four dilution steps. SD analysis was carried out (number of replicates = 11, Supplementary…

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  Figure 2. (A) Direct qPCR amplification with and without nucleic acid extraction for ASFV detection. The mean Ct values based on different FTA cards. SD analysis was carried out (number of replicates = 7, Supplementary Table S2) are shown. Standard deviation (SD) value for all cards by the direct qPCR amplification is 0.78 and for qPCR amplification with extraction is 1.14. The standard error of the mean value for all methods by direct amplification is 0.29 and for the amplification with extraction is 0.43. An unpaired multiple t-test was performed to test the significance between the different FTA cards based on both the qPCR amplification direct and with extraction with a resulting p-value > 0.99 for the taken values, which is not statistically significant. (B) Direct RT-qPCR amplification with and without nucleic acid extraction for IAV detection. The mean Ct values are based on different FTA cards. SD analysis was carried out (number of replicates = 7, Supplementary Table S2) are shown. Standard deviation (SD) value for all cards by the direct RT-qPCR amplification is 0.76 and for RT-qPCR amplification with extraction is 1.74. The standard error of the mean value for all methods by…