Evidence of human coronavirus HKU1 and human bocavirus in Australian children

Abstract

Undiagnosed cases of respiratory tract disease suspected of an infectious aetiology peak during the winter months. Since studies applying molecular diagnostic assays usually report reductions in the number of undiagnosed cases of infectious disease compared to traditional techniques, we applied PCR assays to investigate the role of two recently described viruses, namely human coronavirus (HCoV) HKU1 and human bocavirus (HBoV), in a hospital-based paediatric population. Both viruses were found among Australia children with upper or lower respiratory tract disease during the autumn and winter of 2004, contributing to 21.1% of all microbial diagnoses, with individual incidences of 3.1% (HCoV-HKU1) and 5.6% (HBoV) among 324 specimens. HBoV was found to coincide with another virus in more than half of all instances and displayed a single genetic lineage, whilst HCoV-HKU1 was more likely to occur in the absence of another microbe and strains could be divided into two genetic lineages which we propose be termed HCoV-HKU1 type A and type B. Children under the age of 2 years were most at risk of infection by these viruses which contribute significantly to the microbial burden among patients with respiratory tract disease during the colder months.

ELISAs using human bocavirus VP2 virus-like particles for detection of antibodies against HBoV

“…In addition, a high positive rate of HBoV IgG antibody in sera from healthy U.S. blood donors (aged 18-65 years) was observed in this study, indicating HBoV infection is widespread in the U.S.. Together, these results also indicate that HBoV is likely first encountered in childhood, which is consistent with previous suggestions (Allander et al, 2005;Arnold et al, 2006;Bastien et al, 2006;Choi et al, 2006;Foulongne et al, 2006;Lin et al, 2007;Ma et al, 2006;Qu et al, 2007;Sloots et al, 2006;Weissbrich et al, 2006). Empty VP2 VLPs of B19 have been used effectively as antigen in ELISAs for evaluating B19 antibodies (Soderlund et al, 1995).…”

“…Although IgG antibody present in a single serum sample does not reflect an acute infection, a high rate of IgG positive sera from healthy children does suggest that HBoV infection is more common in children than genomic detection has shown previously (Allander et al, 2005;Arnold et al, 2006;Bastien et al, 2006;Choi et al, 2006;Foulongne et al, 2006;Lin et al, 2007;Ma et al, 2006;Qu et al, 2007;Sloots et al, 2006;Weissbrich et al, 2006). Infection with B19 occurs in children in a high incidence asymptomatically.…”

Human bocavirus 1 and 3 infection in children with acute gastroenteritis in Brazil

“…The viral DNA was extracted using a commercial kit (Wizard Genomic DNA Purification Kit, Promega) according to the manufacturer's instructions. The extracted material was amplified using the primer pairs HBoV 01.2 and HBoV 02.2 (Sloots et al 2006) to specifically target a segment of the NS1 gene of HBoV-1 and HBoV-3. The reaction mixture was prepared for a final volume of 25 µL using 5 x reaction buffer (PCR buffer -20 mM TRIS-HCL pH 8, 4/20 mM KCL 500m mM), 10 mM of each deoxynucleotide triphosphate, 25 mM MgCl 2 , 0.2 µM of each specific primer and 2 U/µL of Taq DNA polymerase (Invitrogen/ Life Technologies).…”

Determination of the Frequency of Human Bocavirus and Other Respiratory Viruses Among 0-2 Years Age Group Children Diagnosed as Acute Bronchiolitis

“…HBoV saptanan örneklerde HBoV DNA'sının gösterilmesi için, prototip HBoV st1 suşu-nun NS1 geninin (1545-1835 nt) 290 baz çift (bç)'lik kısmını hedefleyen NS1F (5'-TAT GGC CAA GGC AAT CGT CCA AG-3') ve NS1R (GCC GCG TGA ACA TGA GAA ACA GA-3') primerleri ile PCR amplifikasyonu gerçekleştirildi 13 . Her bir örneğin PCR amplifikasyonu 50 μl'lik reaksiyon hacimlerinde uygulandı.…”