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. 2013 Oct 18;8(10):e78125. doi: 10.1371/journal.pone.0078125

Figure 4. Influence of swabs from chickens on the real-time PCR assay for receptor binding specificity.

Figure 4

A microplate was immobilized with 200 ng/well of α2,3PGA or α2,6PGA and then blocked with 1% lipid-free BSA in PBS at 4 °C overnight. Avian IAV D313 (A and B) or human IAV M71 (C and D) (2° HAU, 25 μl/well) was mixed with a 5-dilution suspension of a trachea swab (A and C) or cloaca swab (B and D) (25 μl/well) from healthy chickens. The mixtures (50 μl/well) were incubated for 2 h at 4 °C on microplates that were immobilized with α2,3PGA or α2,6PGA. Viral RNAs of viruses bound to α2,3PGA or α2,6PGA were extracted. Viruses that bound to α2,3PGA or α2,6PGA were measured as copy number of HA cDNA by using real-time PCR. The data (S.E.) is the average from three independent experiments. The trachea swab and cloaca swab had no influence on this assay.