Table 1.
List of primers for the assemblage-specific PCRs and sizes of the amplicons.
Table 2.
Selected clones from the library of Giardia duodenalis Ad-28 strain and their genome localizations on both assemblages B (GS) and A (WB).
Figure 1.
Primer design for the 4E1-HP assay.
Alignment of the sequence of the 4E1 clone (assemblage B) with the homologous sequence of HP 13988 (assemblage A). Dots correspond to identical nucleotides. The primers designed for assemblage A amplification are underlined with single lines, whereas the primers designed for assemblage B amplification are underlined with double lines.
Figure 2.
PCR assays at six independent genetic loci.
Locus names are shown at the upper left corner of each panel. Capital letters in the upper lane indicate the reference DNA in each sample: A for assemblage A, B for assemblage B. Neg indicates the negative control (no DNA). Lower cases in the lower lane indicate the specific primer pair used for each reaction: a for the assemblage A specific pair; b for the assemblage B specific pair; a+b for the two primer pairs combined. Sizes of the assemblage-specific bands are indicated on the right. Size markers are indicated on the left.
Figure 3.
PCR amplification of the 4E1-HP and 5C1-P21 markers from 15 human fecal samples positive for G. duodenalis.
Samples 1–5: assemblage A; samples 6–10: assemblage B; samples 11–15: mixed assemblage A+B.
Figure 4.
Sensitivity of 4E1-HP and 5C1-P21 PCR assays.
Panel A: sensitivity test on serial dilutions of reference DNAs (WB for assemblage A and Ad28 for assemblage B). Left images show results obtained with assemblage A; right images show results obtained with assemblage B. The amount of DNA template, reported as the number of cysts equivalent, is shown at the top of the images. Panel B: results of assays tested on different proportions of the same reference DNAs. The following A to B ratios were tested: 6∶4 (1); 7.5∶2.5 (2,); 9∶1 (3); 1∶9 (4); 2.5∶7.5 (5); and 4∶6 (6). N indicates the negative control.
Table 3.
Genotyping results at the 4E1 and 5C1 markers from 51 human samples from Sweden previously characterized by MLG analysis [16].