domingo, 22 de abril de 2012

Enterovirus 104 Infection in Adult, Japan, 2011 - Vol. 18 No. 5 - May 2012 - Emerging Infectious Disease journal - CDC

Enterovirus 104 Infection in Adult, Japan, 2011 - Vol. 18 No. 5 - May 2012 - Emerging Infectious Disease journal - CDC

Volume 18, Number 5—May 2012


Enterovirus 104 Infection in Adult, Japan, 2011

Suggested citation for this article
To the Editor: Human enterovirus (HEV) C (family Picornaviridae, genus Enterovirus) consists of 3 types of poliovirus (1, 2, and 3), 9 types of coxsackievirus A (CV-A1, 11, 13, 17, 19, 20, 21, 22, and 24), and 9 types of enterovirus (EV) (95, 96, 99, 102, 104, 105, 109, 113, and 116)

( EV-104 was first identified in 2009 in Switzerland in 8 children who had pneumonia or acute otitis media (1). To our knowledge, there has been only 1 other report of EV-104, detected in Italy in 3 adults and 2 children who had upper respiratory tract infection (RTI) (2). We report the detection of a novel EV-104 strain in an adult with upper RTI in Japan.

In February 2011, a nasal swab specimen was collected from a 36-year-old immunocompetent man in Japan who had rhinorrhea, cough, pharyngitis, and fever (38.3°C). The sample underwent viral nucleic acid extraction and cDNA synthesis (3) and was PCR screened for HEV and human rhinovirus [HRV] by using primers EVP4 and OL68-1, which detect viral protein (VP) 4/VP2 gene in HEV and HRV as amplicons of ≈650 and 530 bp, respectively (4). Unexpectedly, an amplicon of ≈600 bp was generated.

To identify this amplicon, sequencing and BLAST analysis ( was conducted and yielded a 522-nt sequence with sequence similarity to EV-104 (94.4% identity with the prototype strain CL-12310945; 7,229 nt [GenBank accession no. EU840733]). The sequence similarity corresponded to nt 633–1154 of the novel strain, which was designated AK11 (7,408 nt; GenBank accession no. AB686524).

The complete genome sequence of AK11 was determined as follows: cDNA was synthesized by using sequence-specific primers and amplified as 4 fragments (nt 1–494, 65–3852, 1975–3852, and 3284–7408 with poly A). End-specific nucleotide sequences were determined by using the 5′ RACE system (Rapid Amplification of cDNA Ends; Invitrogen, Life Technologies Corp., Carlsbad, CA, USA) and 3′ RACE by using primer TX30SXN (5).

The genome and predicted amino acid sequences of AK11 were compared with those of CL-12310945, the only EV-104 strain for which a large part of the genome sequence is available. This analysis showed that CL-12310945 is shorter than AK11 at both termini. Specifically, nt 1–64 (corresponding to the 5′ untranslated region [UTR]) and nt 7291–7408 (corresponding to part of the 3D gene and the 3′ UTR) of AK11 were not sequenced from CL-12310945.

The identities between the strains were calculated by using BioEdit version 7.09 (, with results as follows: 5′ UTR (partial sequence 95.0% nt identity; amino acid identity not applicable), VP4 (95.2% nt, 100% aa), VP2 (95.6% nt, 96.3%), VP3 (95.2% nt, 99.6% aa), VP1 (96.2% nt, 99.0% aa), 2A (96.2% nt, 99.3% aa), 2B (94.8% nt, 100% aa), 2C (92.0% nt, 99.1% aa), 3A (83.3% nt, 94.3% aa), 3B (84.8% nt, 90.9% aa), 3C (84.5% nt, 94.5% aa), and 3D (partial sequence 84.7% nt, 93.3% aa). The 3′ UTR was not analyzed.

Thumbnail of Phylogenetic tree of human enterovirus C and viral protein (VP) 1 sequences constructed by using the neighbor-joining method. Complete VP1 gene sequences in enterovirus (EV) 104 (888 nt, corresponding to nt 2461–3348 of novel strain AK11) and other human enterovirus C viruses were aligned by using ClustalX version 2 ( Sequences in EV-95, EV-105, EV-113, and EV-116 were not available from the database. Genetic distances between sequences were calculated by using the Figure. . . Phylogenetic tree of human enterovirus C and viral protein (VP) 1 sequences constructed by using the neighbor-joining method. Complete VP1 gene sequences in enterovirus (EV) 104 (888 nt, corresponding...

Phylogenetic analysis of VP1 sequences among HEV-C viruses showed that AK11 clusters with CL-12310945 and is genetically close to CV-A1, CV-A19, CV-A22, and EV-109 (Figure). These results are consistent with reported results (6). Virus isolation, attempted by using Vero and RD-18S cells, was unsuccessful. This result is consistent with previous EV-104 reports, wherein the virus could not be grown or isolated (1,2).
To determine the presence of other respiratory viruses in this patient, the EV-104–positive specimen was tested by using real-time PCR for any of 17 other viruses (human metapneumovirus, respiratory syncytial virus, human parainfluenza virus types 1–4, human bocavirus, human coronavirus [229E, OC43, HKU1, NL63], influenza virus [A, pandemic (H1N1) 2009, B, C], human adenovirus, and HRV). No other viruses were detected (data not shown). This result indicates that EV-104 was associated with upper RTI in this patient. During the 2 months in which the EV-104–positive sample was collected, influenza A virus, HRV, and respiratory syncytial virus were most frequently detected in other patients, and no enterovirus was observed in other specimens from persons with RTI.
EV-104 detection is rare (5/1,500 [0.3%] for a 1-year study in Italy [1]; 8/1,592 [0.5%] for a 10-year study in Switzerland [2]). As part of a virus surveillance program in Osaka City, Japan, during November 2010–October 2011, a total of 645 respiratory tract specimens were collected from children with RTI (360 male, 285 female; age 0–59 months, mean ± SD 18.9 ± 13.8 months) and subjected to PCR by using EVP4 and OL68-1 primers. No EV-104 was detected. In 2 previous studies in Japan, we detected no EV-104 in 764 specimens from patients with RTI during November 2008 and October 2010 (3,7); therefore, we have found EV-104 in only 1 (0.07%) of 1,410 samples tested.
Infrequent detection and insensitivity to cell culture contribute to the rarity of EV-104 identification. However, given the lack of contact between EV-104–positive patients in Italy and Switzerland, more RTI patients might actually be carrying EV-104 than testing has indicated. The collection of additional EV-104 strains and associated epidemiologic and virologic information will help clarify the role of this virus in RTI.
Atsushi KaidaComments to Author , Hideyuki Kubo, Jun-ichiro Sekiguchi, Atsushi Hase, and Nobuhiro Iritani
Author affiliations: Osaka City Institute of Public Health and Environmental Sciences, Osaka, Japan


This work was supported in part by a grant-in-aid for Young Scientists (B) (23790720) from The Ministry of Education, Culture, Sports, Science, and Technology of Japan.


  1. Tapparel C, Junier T, Gerlach D, Van-Belle S, Turin L, Cordey S, New respiratory enterovirus and recombinant rhinoviruses among circulating picornaviruses. Emerg Infect Dis. 2009;15:71926. DOIExternal Web Site IconPubMedExternal Web Site Icon
  2. Piralla A, Rovida F, Baldanti F, Gerna G. Enterovirus genotype EV-104 in humans, Italy, 2008–2009. Emerg Infect Dis. 2010;16:101821.PubMedExternal Web Site Icon
  3. Kaida A, Kubo H, Sekiguchi J, Kohdera U, Togawa M, Shiomi M, Enterovirus 68 in children with acute respiratory tract infections, Osaka, Japan. Emerg Infect Dis. 2011;17:14947.PubMedExternal Web Site Icon
  4. Ishiko H, Miura R, Shimada Y, Hayashi A, Nakajima H, Yamazaki S, Human rhinovirus 87 identified as human enterovirus 68 by VP4-based molecular diagnosis. Intervirology. 2002;45:13641. DOIExternal Web Site IconPubMedExternal Web Site Icon
  5. Katayama K, Shirato-Horikoshi H, Kojima S, Kageyama T, Oka T, Hoshino F, Phylogenetic analysis of the complete genome of 18 Norwalk-like viruses. Virology. 2002;299:22539. DOIExternal Web Site IconPubMedExternal Web Site Icon
  6. Yozwiak NL, Skewes-Cox P, Gordon A, Saborio S, Kuan G, Balmaseda A, Human enterovirus 109: a novel interspecies recombinant enterovirus isolated from a case of acute pediatric respiratory illness in Nicaragua. J Virol. 2010;84:904758. DOIExternal Web Site IconPubMedExternal Web Site Icon
  7. Kaida A, Kubo H, Takakura K, Togawa M, Shiomi M, Kohdera U, Molecular epidemiology of human rhinovirus C in patients with acute respiratory tract infections in Osaka City, Japan. Jpn J Infect Dis. 2011;64:48892.PubMedExternal Web Site Icon


Suggested citation for this article: Kaida A, Kubo H, Sekiguchi J, Hase A, Iritani N. Enterovirus 104 infection in adult, Japan, 2011 [letter]. Emerg Infect Dis [serial on the internet]. 2012 May [date cited]. Web Site Icon
DOI: 10.3201/eid1805.111890

No hay comentarios:

Publicar un comentario