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Figure 1 | BMC Infectious Diseases

Figure 1

From: Norovirus-related chronic diarrhea in a patient treated with alemtuzumab for chronic lymphocytic leukemia

Figure 1

Evolution of norovirus (NoV) loads and number of stools after alemtuzumab treatment. NoV loads are represented in black dashed lines and expressed in log copies per g of stool. Stools are represented in gray solid lines and expressed in number per day. First, fecal samples have been weighed and then suspended in phosphate-buffered saline (pH 7.5) at a concentration of 10%. Viral RNA was extracted with a QIAamp viral RNA kit (Qiagen, Hilden, Germany). The NoVs were detected by RT-PCR on an ABI Prism 7500 Fast detector (Life Technologies) using the TaqMan One-Step PCR Master Mix reagent (Life Technologies) and previously published primers and probes for GI [18] and GII [10]. For quantitative assessment of fecal viral load, the number of NoV RNA copies was estimated by comparing the sample CT value with standard curves. Among the five available samples, four were positive for GII.7 in the polymerase gene and for GII.6 in the capsid gene. Then, as described previously [10], the ORF1/ORF2 junction was sequenced using the same primers. The nucleotide sequences were aligned and compared to corresponding sequences of NoV strains available in the GenBank database using Fasta program. These sequences showed a homology of 92% and 98% for GII.7 and GII.6, respectively, and a 100% homology between them. Abbreviations: Ig, immunoglobulins; IV, intravenous; NoV, norovirus; W, week.

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