Detection of human bocavirus mRNA in respiratory secretions correlates with high viral load and concurrent diarrhea

Abstract

Human bocavirus (HBoV) is a parvovirus recently identified in association with acute respiratory infections (ARI). Despite its worldwide occurrence, little is known on the pathogenesis of HBoV infections. In addition, few systematic studies of HBoV in ARI have been conducted in Latin America. Therefore, in order to test whether active viral replication of human bocavirus is associated with respiratory diseases and to understand the clinical impact of this virus in patients with these diseases, we performed a 3-year retrospective hospital-based study of HBoV in outpatients and inpatients with symptoms of Acute Respiratory Infections (ARI) in Brazil. Nasopharyngeal aspirates (NPAs) from 1015 patients with respiratory symptoms were tested for HBoV DNA by PCR. All samples positive for HBoV were tested by PCR for all other respiratory viruses, had HBoV viral loads determined by quantitative real time PCR and, when possible, were tested by RT-PCR for HBoV VP1 mRNA, as evidence of active viral replication. HBoV was detected in 4.8% of patients, with annual rates of 10.0%, 3.0% and 3.0% in 2005, 2006 and 2007, respectively. The range of respiratory symptoms was similar between HBoV-positive and HBoV-negative ARI patients. However, a higher rate of diarrhea was observed in HBoV-positive patients. High HBoV viral loads (>10⁸ copies/mL) and diarrhea were significantly more frequent in patients with exclusive infection by HBoV and in patients with detection of HBoV VP1 mRNA than in patients with viral co-infection, detected in 72.9% of patients with HBoV. In summary, our data demonstrated that active HBoV replication was detected in a small percentage of patients with ARI and was correlated with concurrent diarrhea and lack of other viral co-infections.

Figure 1. Monthly distribution of samples positive for HBoV and HRSV in the years 2005 through 2007 in Ribeirão Preto, Brazil in relation to monthly accumulated rainfall and average temperauture (http://www.ciiagro.sp.gov.br).

Figure 2. Viral loads of HBoV (copies of genomic DNA/mL) in NPAs by qPCR.

A. HBoV viral loads in NPAs from patients with HBoV as single agent (black circles), and with simultaneous detection of other respiratory viruses (white circles). B. HBoV viral loads in NPAs from patients with (black circles) and without (white circles) detectable shedding of HBoV VP1 mRNA by real time PCR.