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. 2013 Apr;19(4):574-80.
doi: 10.3201/eid1904.121775.

Detection of spliced mRNA from human bocavirus 1 in clinical samples from children with respiratory tract infections

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Detection of spliced mRNA from human bocavirus 1 in clinical samples from children with respiratory tract infections

Andreas Christensen et al. Emerg Infect Dis. 2013 Apr.

Abstract

Human bocavirus 1 (HBoV1) is a parvovirus associated with respiratory tract infections (RTIs) in children, but a causal relation has not yet been confirmed. To develop a qualitative reverse transcription PCR to detect spliced mRNA from HBoV1 and to determine whether HBoV1 mRNA correlated better with RTIs than did HBoV1 DNA, we used samples from HBoV1 DNA-positive children, with and without RTIs, to evaluate the test. A real-time reverse transcription PCR, targeting 2 alternatively spliced mRNAs, was developed. HBoV1 mRNA was detected in nasopharyngeal aspirates from 33 (25%) of 133 children with RTIs but in none of 28 controls (p<0.001). The analytical sensitivity and specificity of the test were good. Our data support the hypothesis that HBoV1 may cause RTIs, and we propose that HBoV1 mRNA could be used with benefit, instead of HBoV1 DNA, as a diagnostic target.

Keywords: Human bocavirus; children; mRNA; parvovirus; polymerase chain reaction; respiratory tract infections; splicing; viruses.

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Figures

Figure 1
Figure 1
Schematic representation of the 2 human bocavirus 1 (HBoV1) mRNA PCR products, illustrating alternative splicing. Positions of primers and probe are shown. The total length of the upper product is 242 bp, and the length of the lower is 363 bp (reference sequence: GenBank accession no. NC007455).
Figure 2
Figure 2
Agarose gel stained with ethidium bromide. Reverse transcription PCR products from 4 patients are shown in lanes 2–5 and 1-kb DNA Ladder (Life Technologies Corp., Carlsbad, CA, USA) in lane 1. Arrows indicate 2 bands corresponding to ≈250 and ≈400 bp.
Figure 3
Figure 3
Distribution of human bocavirus 1 (HBoV1) DNA loads in nasopharyngeal aspirates either positive (n = 33) or negative (n = 128) for HBoV1 mRNA. Each dot indicates 1 sample.

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