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Localisation and length of B19 amplicons

This method involves quantifying amplicons, assessing inhibition efficacy by adjusting pre-amplification cycles, analyzing adduct formation, and comparing DNA/virion inactivation. Explore the effects of amplicon lengths and exposure time, with detailed protocols.

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Localisation and length of B19 amplicons

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  1. Localisation and length of B19 amplicons

  2. Principle of the method of quantification Adduct formation is a random phenomenon related to S59 concentration The longer the amplicon the more chance of being inhibited Assessing efficacy of inhibition by pre-amplifying amplicons of different length and measuring amplified template by short QPCR Pre-amplification number of cycle adjusted to provide approximately same number of templates: 244 nt 8 cycles 926 nt 10 cycles quantification by 3011 nt 16 cycles QPCR 5300 nt 30 cycles

  3. Inhibition of HEV amplification according to amplicon length and time of exposure to S19 3 2.5 5 min 2 15 min 30 min Log inhibition 1.5 45 min 60 min 75 min 1 90 min 0.5 0 80 250 900 2500 5000 Amplicon length

  4. B19-Inhibition for different strains 3 2.5 127 2 131 1.5 129 log inhibition 1 133 0.5 0 80 250 900 2500 5000 length

  5. Comparison of B19 DNA or virion inactivation with S59 Sample 2 7 6 5 4 DNA Log inhibition of amplification Virions 3 2 1 0 0 1000 2000 3000 4000 5000 6000 Amplified genome length

  6. Comparison of B19 DNA or virions inactivation with S59 Sample 1 4.5 4 3.5 3 2.5 DNA Log inhibition of amplification Virions 2 1.5 1 0.5 0 0 1000 2000 3000 4000 5000 6000 Length of genome amplified

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