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. 2016 Nov 8;6:234. doi: 10.3389/fonc.2016.00234

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Copyright © 2016 Wu, Park, Deighan, Amaya, Miller, Pan, Zborowski, Lustberg and Chalmers.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Integrated RNA ISH and ICC staining on HPV markers. Integrated RNA ISH and ICC staining with customized conjugated CK8, 18, 19-AF488 antibody (ICC), HPV16 E6, E7-type 1 (RNA ISH), and EGFR-AF647 antibody (ICC). The HPV16 E6, E7-type 1 probe is the 1:1 mixture of HPV16 E6-type 1 and HPV16 E7-type 1. (A,B) presents the staining of blood cells spiked with SCC90 (A) or SCC47 (B) cells at a ratio of 4:1 in cell number. (C) shows the staining of the NME cells from head and neck cancer patients. Among these NME cells shown in (C), HPV+, EGFR+, CK+, or CK− cells are found. (D) is a specificity test with the staining of blood cells spiked with BT474 cells at a ratio of 4:1 in cell number. Both BT474 cells and blood cells are negative on HPV. The specificity of this test is hence proved.

Integrated RNA ISH and ICC staining on HPV markers. Integrated RNA ISH and ICC staining with customized conjugated CK8, 18, 19-AF488 antibody (ICC), HPV16 E6, E7-type 1 (RNA ISH), and EGFR-AF647 antibody (ICC). The HPV16 E6, E7-type 1 probe is the 1:1 mixture of HPV16 E6-type 1 and HPV16 E7-type 1. (A,B) presents the staining of blood cells spiked with SCC90 (A) or SCC47 (B) cells at a ratio of 4:1 in cell number. (C) shows the staining of the NME cells from head and neck cancer patients. Among these NME cells shown in (C), HPV+, EGFR+, CK+, or CK− cells are found. (D) is a specificity test with the staining of blood cells spiked with BT474 cells at a ratio of 4:1 in cell number. Both BT474 cells and blood cells are negative on HPV. The specificity of this test is hence proved.