Norovirus RNA in serum associated with increased fecal viral load in children: Detection, quantification and molecular analysis

Abstract

Worldwide, norovirus (NoV) is a major cause of acute gastroenteritis (AGE) responsible for pandemics every ~3 years, and over 200,000 deaths per year, with the majority in children from developing countries. We investigate the incidence of NoV in children hospitalized with AGE from Belém, Pará, Brazil, and also correlated viral RNA levels in their blood and stool with clinical severity. For this purpose, paired stool and serum samples were collected from 445 pediatric patients, ≤9 years between March 2012 and June 2015. Enzyme-linked immunosorbent assay (EIA) was used to detect NoV in stool and reverse transcription quantitative PCR (RT-qPCR) used to quantify NoV RNA levels in sera (RNAemia) and in the positive stool. Positives samples were characterized by the partial ORF1/2 region sequence of viral genome. NoV antigen was detected in 24.3% (108/445) of stool samples, with RNAemia also present in 20.4% (22/108). RNAemia and a high stool viral load (>107 genome copies/gram of faeces) were associated with longer hospitalizations. The prevalent genotypes were GII.4 Sydney_2012 (71.6%-58/81) and New Orleans_2009 (6.2%-5/81) variants. Eight other genotypes belonging to GII were detected and four of them were recombinant strains. All sera were characterized as GII.4 and shared 100% similarity with their stool. The results suggest that the dissemination of NoV to the blood stream is not uncommon and may be related to increased faecal viral loads and disease severity.

Detection and genotyping of enteric viruses in hospitalized children with acute gastroenteritis in Belém, Brazil: Occurrence of adenovirus viremia by species F, types 40/41

“…All samples (stools and sera) were collected as soon as possible after hospitalization and sent to the Evandro Chagas Institute, where they were processed and kept frozen (−20°C) until use. Initially, they were tested for RV and NoV by enzyme immunoassay, and only samples with negative results (N = 219 paired samples) were selected for current study. Serum and fecal samples were analyzed following the same methodology of detection, as described below.…”

Evolutionary and Molecular Analysis of Complete Genome Sequences of Norovirus From Brazil: Emerging Recombinant Strain GII.P16/GII.4

“…This study on the emergence of the GII.P16/GII.4 strain started from previous data from studies conducted with samples from the cities Belém and Manaus in northern Brazil (Hernandez et al, 2018;Reymão et al, 2018).…”

“…GII.4 was the genotype most frequently detected, with the emergence of the GII.P31/GII.4-Sydney 2012, GII.17_2014 and GII.P16/GII.4 strains. Reports about GII.P31/GII.4 and GII.17_2014, as well as the temporal distribution and emergence period of these strains, have already been described (Silva et al, 2013;Hernandez et al, 2018;Reymão et al, 2018;Costa et al, 2019).…”

“…0017/2014 update No. 1.318.103 of 2015) as described by Hernandez et al (2018) and Reymão et al (2018).…”

Development of CdSe–ZnO Flower-Rod Core-Shell Structure Based Photoelectrochemical Biosensor for Detection of Norovirous RNA

“…There is a high incidence rate, so it is very important to develop rapid and accurate detection techniques for the prevention and control of the transmission of norovirus. Conventional detection methods of NV RNA include immunochromatography [ 3 ] and reverse transcription-polymerase chain reaction (RT-PCR) [ 4 , 5 ]. However, they have certain limitations, such as contrasting sensitivities, post-amplification specimen handling, assay complexity, and the requirement for accurate performance tests [ 6 ].…”