Norwalk virus shedding after experimental human infection

Abstract

Noroviruses are the most common cause of viral gastroenteritis in the United States. To determine the magnitude and duration of virus shedding in feces, we evaluated persons who had been experimentally infected with Norwalk virus. Of 16 persons, clinical gastroenteritis (watery diarrhea and/or vomiting) developed in 11; symptomatic illness lasted 1-2 days. Virus shedding was first detected by reverse transcription-PCR (RT-PCR) 18 hours after participant inoculation and lasted a median of 28 days after inoculation (range 13-56 days). The median peak amount of virus shedding was 95 x 10(9) (range 0.5-1,640 x 10(9)) genomic copies/g feces as measured by quantitative RT-PCR. Virus shedding was first detected by antigen ELISA approximately 33 hours (median 42 hours) after inoculation and lasted 10 days (median 7 days) after inoculation. Understanding of the relevance of prolonged fecal norovirus excretion must await the development of sensitive methods to measure virus infectivity.

Figure 1

Shedding of Norwalk virus in feces. The quantity of viral RNA measured by quantitative reverse transcription–PCR (qRT-PCR; black line) and of virus antigen measured by ELISA (optical density; blue line) is shown for 2 participants: no. 703, who did not have clinical gastroenteritis (panel A), and no. 721, who had clinical gastroenteritis (panel B). Vertical lines represent the period of clinical symptoms; N, nausea; V, vomiting. Panels C, D, and E show the virus titers as measured by qRT-PCR in fecal samples collected from participants who had no clinical gastroenteritis, had gastroenteritis with vomiting only, and had gastroenteritis with vomiting and diarrhea, respectively.

Figure 2

Correlation of viral RNA titer with antigen ELISA (optical density [OD]). Titers of viral RNA are correlated with the OD measured by antigen ELISA for the 148 fecal samples with positive quantitative reverse transcription–PCR (qRT-PCR) results (r² = 0.823, Spearman correlation, p<0.001).