Additional file 2 is a schematic representation of the different possible outcomes in the event of an assemblage B Giardia infection. Moreover, the data presented here strongly highlights the necessity of re-evaluating the current molecular epidemiological methods used for sub-genotyping of assemblage B Giardia. The concurrence of ASH at the EX 527 datasheet single cell level, and the seemingly high frequency of mixed sub-genotype infections in clinical samples makes it profoundly difficult to verify specific assemblage B sub-genotypes in clinical samples, using the current genotyping tools. Acknowledgements This study was sponsored by grants
from SIDA/SAREC, The Swedish Medical Research Council (VR-M) and Formas. selleck chemicals We thank Görel Allestam for technical assistance. We also thank Professor Mats Wahlgren for generously providing us access to his micromanipulator. Electronic supplementary material Additional file 1: Single Giardia cells were isolated by micromanipulation, using micro capillaries with a 6 – 8 μm inner diameter (panel A). Picked cells were transferred to a 2 μl pure drop of 1X PBS for re-verification (panel B), and subsequently transferred to the PCR reaction mixture. (PPT 2 MB) Additional file 2: A schematic representation of a mixed infection, where the red and blue bars represent different alleles of the same gene in different G. intestinalis sub-assemblages (a), and a single parasite harboring ASH, where red and blue bars indicate different
alleles of the same gene within a single cell (b). This is a simplistic, schematic representation of different almost modes of infection in a giardiasis patient with parasites of different assemblage B sub-assemblages, bringing forth the topics addressed in this study where mixed infection of different sub-assemblages, the occurrence of ASH in a clonal Giardia strain, or a mixture of the two may be present in a patient. Thus selleck chemicals llc highlighting an important biological phenomenon in
Giardia, as well as suggesting a revision of the current strategy used in assemblage B Giardia epidemiology. (PPT 160 KB) References 1. Lasek-Nesselquist E, Welch DM, Sogin ML: The identification of a newGiardia duodenalisassemblage in marine vertebrates and a preliminary analysis ofG. duodenalispopulation biology in marine systems. Int J Parasitol 2010,40(9):1063–1074.PubMedCrossRef 2. Ankarklev J, Jerlstrom-Hultqvist J, Ringqvist E, Troell K, Svard SG: Behind the smile: cell biology and disease mechanisms ofGiardiaspecies. Nat Rev Microbiol 2010,8(6):413–422.PubMed 3. Bernander R, Palm JE, Svard SG: Genome ploidy in different stages of theGiardia lamblialife cycle. Cell Microbiol 2001,3(1):55–62.PubMedCrossRef 4. Caccio SM, Ryan U: Molecular epidemiology of giardiasis. Mol Biochem Parasitol 2008,160(2):75–80.PubMedCrossRef 5. Lebbad M, Ankarklev J, Tellez A, Leiva B, Andersson JO, Svard S: Dominance ofGiardiaassemblage B in Leon, Nicaragua. Acta Trop 2008,106(1):44–53.PubMedCrossRef 6.