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In vitro evaluation of primer specificity and comparison of protocols’ sensitivity. Red blocks represent serial dilution that provided positive results in all repetitions. The higher dilution at which all the replicas yielded positive results identified the LOD of the test. Red/white patterns show cases in which even lower dilution resulted in weak positivity or inconsistent results among the different repetitions. † Antigen suspension with initial titer 9 ± 1 log2 HA.
