Human Bocavirus DNA in Paranasal Sinus Mucosa

LETTERS

7. Allander T, Tammi MT, Eriksson M, sinusitis for respiratory viruses and rhinoviruses; human enteroviruses Bjerkner A, Tiveljung-Lindell A, An- atypical bacteria. and parechoviruses; and adenoviruses. dersson B. Cloning of a human parvo- virus by molecular screening of respira- A total of 102 tissue samples Mycoplasma pneumoniae, Chlamydia tory tract samples. Proc Natl Acad Sci were obtained from 88 patients pneumoniae, Legionella pneumophila, U S A. 2005;102:12891–6. doi:10.1073/ (median age 48.5 years, range 13.3– and Bordetella pertussis were analyzed pnas.0504666102 88.1 years) from July 2009 through as described (6–8). 8. Liang Z, Kumar AS, Jones MS, Knowles NJ, Lipton HL. Phylogenetic analy- September 2010 after elective surgery. A single virus was detected in sis of the species Theilovirus: emerg- Indication for surgery was established 22/102 (21.5%) tissue specimens, ing murine and human pathogens. J by otorhinolaryngologists. The most with HBoV being the most frequent Virol. 2008;82:11545–54. doi:10.1128/ common indication was chronic (18/102, 17.6%), followed by JVI.01160-08 9. Chua KB, Khairullah NS, Hooi PS. Sero- sinusitis. No patients displayed acute rhinovirus (2/102, 1.9%), coronavirus epidemiology of human herpesvirus 6 in a respiratory symptoms at the time 229E, and infl uenza A pandemic population seen in the University Hospital, of surgery. To detect asymptomatic (H1N1) 2009 virus (1/102, 0.9%). Kuala Lumpur, Malaysia. Southeast Asian shedding in the upper respiratory HBoV was detected in specimens J Trop Med Public Health. 1996;27:91–5. tract and viremia, we collected nasal collected during July–September Address for correspondence: Lin-Fa Wang, swabs and EDTA-blood samples (13/18) and during February and CSIRO Livestock Industries, Australian Animal concurrently. The study protocol was March (5/18). All positive results Health Laboratory, Geelong, Victoria, Australia; approved by the Ethics Committee of were confi rmed by single real-time email: [email protected] the University of Freiburg. Informed PCR (9). For 14 patients, 2 different written consent was obtained from all mucosal samples were tested and study participants. gave identical results. No multiple Approximately 25 mg of each viral infections and no bacteria were tissue specimen was used for nucleic detected. Median patient age was acid extraction by using an RNeasy 51.2 years (range 14.4–74.2 years) for Mini Kit, as described (5) (QIAGEN, HBoV-positive and 47.6 years (range Hamburg, Germany). To provide 13.3–88.1 years) for HBoV-negative

Human Bocavirus evidence that the QIAGEN RNeasy samples. Ct analysis in single real- kit is also suitable for DNA extraction, time PCR revealed a median C of DNA in Paranasal t we spiked HBoV negative samples 31 (range 28–38), corresponding to Sinus Mucosa with different amounts of HBoV DNA 200 genome equivalents/106 cells 4 6 To the Editor: Human bocavirus before extraction of nucleic acids was (range 3–1.8 × 10 copies/10 cells). done with either the QIAGEN RNeasy No correlation between C value (HBoV) is a newly described parvovirus t for which pathogenic potential has Kit or DNA Blood Kit (QIAGEN). and patients’ age was observed 2 not clearly been elucidated (1). Extracted nucleic acids were then (r = 0.008; data not shown). No Recent fi ndings suggest that HBoV subjected to real-time PCR by using underlying disease was diagnosed may establish persistent infection of primers specifi c for HBoV. Minimal for 13/18 HBoV-positive patients, mucosal lymphocytes or contribute differences (±1 cycle threshold whereas 2/18 and 3/18 patients had [C ] value) in the HBoV PCR were chronic obstructive pulmonary and to tonsillar hyperplasia in children t (2). In previous reports, we described detected; the QIAGEN RNeasy Kit oncologic disease, respectively. prolonged HBoV DNA detection in was therefore used throughout the Nasal swabs and EDTA-blood immunocompromised children (3,4). study (data not shown). Nasal swabs samples were obtained from 17/18 Partial sequencing of the VP1 gene of and EDTA-blood were purifi ed by and 7/18 HBoV-positive patients, HBoV from bronchoalveolar lavage using a QIAamp MinElute Virus respectively. No HBoV was detected fl uid, plasma, and sphenoid sinus Spin Kit (QIAGEN). Multiplex PCR in any swabs or EDTA-blood samples samples showed 100% identity, which for respiratory viruses (Fast-track available, indicating no virus shedding suggested persistence of the same Diagnostics, Junglinster, Luxembourg) in the respiratory tract and no HBoV strain over a 5-month period was conducted to detect infl uenza A viremia. However, the 2 patients with (3). It remains speculative, however, (including pandemic [H1N1] 2009) rhinovirus-positive samples obtained whether paranasal sinus mucosa and B viruses; respiratory syncytial from biopsy also had rhinovirus RNA represents a site of HBoV persistence. virus; human metapneumovirus; detectable in nasal swabs, suggesting To clarify this, we analyzed samples HBoV; parainfl uenza virus 1–4; rhinovirus infection. Unfortunately, of paranasal mucosal tissue and nasal human coronaviruses HKU1, no nasal swab was available from the polyps from patients with chronic NL63, 229E, and OC43; human 2 patients whose sinus biopsy samples

1564 Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 17, No. 8, August 2011 LETTERS were positive for HCoV 229E and Author affi liation: Freiburg University Address for correspondence: Valeria Falcone, infl uenza A virus. Medical Center, Freiburg, Germany Department of Virology, Freiburg University In this study, we simultaneously Medical Center, Hermann-Herder-Strasse 11, DOI: 10.3201/eid1708.101944 analyzed tissue specimens of paranasal 79104 Freiburg, Germany; email: valeria. sinuses and nasal polyps, as well as [email protected] References nasal swabs and blood samples, for a broad panel of viruses and atypical 1. Chow BD, Esper FP. The human bocavi- bacteria. To avoid seasonal bias, ruses: a review and discussion of their role specimens were collected over a 1-year in infection. Clin Lab Med. 2009;29:695– 713. doi:10.1016/j.cll.2009.07.010 period and exclusively obtained from 2. Lu X, Gooding LR, Erdman DD. Human patients undergoing elective surgery bocavirus in tonsillar lymphocytes. Emerg in the absence of acute respiratory Infect Dis. 2008;14:1332–4. doi:10.3201/ symptoms. eid1408.080300 Mixed Genotype 3. Schenk T, Maier B, Hufnagel M, Strahm The fi nding that HBoV was B, Kontny U, Neumann-Haefelin D, et Infections with present as a single virus in 18/22 virus- al. Persistence of human bocavirus DNA Hepatitis C Virus, positive biopsy samples is intriguing. in immunocompromised children. Pediatr Moreover, the fact that no HBoV DNA Infect Dis J. 2011;30:82–4. Pakistan 4. Schenk T, Strahm B, Kontny U, Hufna- was detected in nasal swabs or EDTA- gel M, Neumann-Haefelin D, Falcone V. To the Editor: The prevalence of blood samples indicates no active Disseminated bocavirus infection after hepatitis C virus (HCV) infection is HBoV infection. In previous studies, stem cell transplant. Emerg Infect Dis. high (8% of the population) in Pakistan HBoV DNA was frequently identifi ed 2007;13:1425–7. 5. Herberhold S, Eis-Hübinger AM, Pan- (1). HCV is an RNA virus that has a in the adenoids and tonsils of children ning M. Frequent detection of respiratory high mutation rate. This high rate results (2,5,10). However, in contrast with viruses by real-time PCR in adenoid sam- in extensive genetic heterogeneity, our fi ndings, detection of HBoV was ples from asymptomatic children. J Clin and HCV isolates are found as either mostly associated with other viruses, Microbiol. 2009;47:2682–3. doi:10.1128/ JCM.00899-09 quasispecies or genotypes (2). Humans suggesting that co-virus–induced 6. Welti M, Jaton K, Altwegg M, Sahli R, can be co-infected with >1 genotype cellular damage might contribute Wenger A, Bille J. Development of a mul- (mixed genotype infection) of this to bocavirus reactivation and tiplex real-time quantitative PCR assay to virus (3). The rate of HCV mixed replication (5). Our fi ndings indicate detect Chlamydia pneumoniae, Legionella pneumophila and Mycoplasma pneumoni- genotype infections is extremely that persistence of viral nucleic acid ae in respiratory tract secretions. Diagn variable for different regions and for in sinus mucosa might be a special Microbiol Infect Dis. 2003;45:85–95. the same group of patients tested by advantage of HBoV, although the doi:10.1016/S0732-8893(02)00484-4 using different assays (4). Thus, it is relevance of this observation remains 7. Probert WS, Ely J, Schrader K, Atwell J, Nossoff A, Kwan S. Identifi cation diffi cult to determine the prevalence of unclear. Whether this presence as and evaluation of new target sequences mixed genotype infections by currently a single virus means a dead end for specifi c detection of Bordetella per- available assays, including direct DNA for HBoV infection, true latency tussis by real-time PCR. J Clin Micro- sequencing, because they are designed including the potential of reactivation, biol. 2008;46:3228–31. doi:10.1128/ JCM.00386-08 to identify only the HCV genotype or a role in the pathogenesis of clinical 8. Dumke R, Schurwanz N, Lenz M, Sch- dominant in that particular population. conditions requiring surgery warrants uppler M, Lück C, Jacobs E. Sensitive Consequently, genotypes present at future studies. detection of Mycoplasma pneumoniae in lower frequencies could be missed or human respiratory tract samples by opti- mized real-time PCR approach. J Clin Mi- mistyped (5). Acknowledgment crobiol. 2007;45:2726–30. doi:10.1128/ To determine the prevalence of We are grateful to Gudrun Woywodt JCM.00321-07 HCV mixed genotype infections, we for excellent technical assistance. 9. Neske F, Blessing K, Tollmann F, Schubert retrospectively analyzed genotyping J, Rethwilm A, Kreth HW, et al. Real- This work was supported by the time PCR for diagnosis of human boca- data for paired serum samples from Federal Ministry of Education and virus infections and phylogenetic analy- 22,125 HCV-infected patients during Research (contract no. 01ES0830). sis. J Clin Microbiol. 2007;45:2116–22. the past 11 years (March 2000–May doi:10.1128/JCM.00027-07 2010) for all regions in Pakistan by 10. Sato M, Li H, Ikizler MR, Werkhaven JA, Valeria Falcone, Gerd J. Ridder, Williams JV, Chappell JD, et al. Detection using molecular-based genotype- Marcus Panning, of viruses in human adenoid tissues by specifi c methods (6,7). A total of Sibylle Bierbaum, use of multiplex PCR. J Clin Microbiol. 12,036 (54.4%) were male patients 2009;47:771–3. doi:10.1128/JCM.02331- and 10,089 (45.6%) were female Dieter Neumann-Haefelin, 08 and Daniela Huzly patients.

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