We also employed a blast colony assay (Kennedy et al., 1997) to show that the development of the earliest stage of hematopoietic development, the hemangioblast, was not disturbed and similar numbers of colonies were formed (supplementary material Fig. S2C).
We have utilized the blast cell assay of Buick et al. (1977) to grow and subsequently cytogenetically analyze cultured colony forming cells (CFUs) from patients with acute and chronic myelogenous leukaemia (AML, CML).
Blast colony assays were performed as previously described [16].
Unlike previous studies[3], our blast colony assays were performed on EB6 cells instead of EB3.0 or 3.5 cells.
Blast colony assays revealed that most hemangioblast activity was detected apart from the pre-HSC population, implying the early divergence of pre-HSCs from hemangioblasts.
The blast cell colony assay, similar to the HPP-CFC assay, is another variant of the CFC assay that detects cells with an extensive proliferative capacity based, in part, on their potential to generate secondary colonies when replated (14).
For these 21 A/H5N1-positive specimens, the BLAST analyses of assay-generated M, NS and PB2 gene sequences exclusively represented subtype A/H5N1 avian influenza viruses, as 255 M gene, 146 NS gene and 310 PB2 gene sequence records.
Table 1 shows that the most similar sequence records returned from BLAST analysis of assay-generated HA1, NA1, HA3, NA2, M H1N1), M H3N2) gene sequences, as well as type B HA, NA and M gene sequences, all correctly correspond to the actual influenza virus strains used to represent these components in the 2004 2005 vaccine configuration.
In the present study, 63% of the positive samples detected in the hexon region were also positive using the fiber assay, and BLAST analysis was concordant with the hexon analysis for these sequences.
Conceptual translation of Irm revealed that it is a noncoding RNA as it only has several short ORFs that lack significant homology to any known protein as assayed by blast searches (data not shown).
In marked contrast to results from RPM-Flu assay-generated HA and NA gene sequences, there was poor correspondence of the actual specimen A/HN subtypes with the subtypes of sequence records returned from BLAST analysis of the assay-generated M, and NS gene sequences.
