Skip directly to site content Skip directly to page options Skip directly to A-Z link Skip directly to A-Z link Skip directly to A-Z link
Volume 21, Number 8—August 2015

Avian Gyrovirus 2 DNA in Fowl from Live Poultry Markets and in Healthy Humans, China

On This Page
Article Metrics
citations of this article
EID Journal Metrics on Scopus

Cite This Article

To the Editor: In 2011, a chicken anemia virus (CAV)–related sequence, designated avian gyrovirus 2 (AGV2), was first identified in serum samples from diseased chickens in Brazil (1). During the same year, a human gyrovirus (HGyV) sequence that had high identity to AGV2 was detected in the skin of humans in France (2). As with CAV, 3 open reading frames (ORFs) for encoding viral proteins (VP) 1–3 (2) overlapped in genome of AGV2. Recently, HGyV/AGV2 has been detected in Hong Kong in chicken meat for consumption by humans, in human blood samples from donors in France, and in HIV-positive persons and organ transplant recipients in Italy and the United States (35). However, the epidemiology, host range, transmission route, and pathogenesis of AGV2 remain poorly understood. Bullenkamp et al. found that AGV2 VP3 protein, like CAV VP3, can induce apoptosis of tumor cells (6). Also, Abolnik et al. reported the detection in Southern Africa of AGV2 in brain tissue of chickens that showed severe neurologic signs (7). These findings highlight the potential pathogenesis of AGV2.

So far, little is known about AGV2 in mainland China among chickens and humans. Because live poultry market (LPMs) play a critical role in the transmission of poultry pathogens to humans, we used PCR to investigate the presence of AGV2 in chickens (54 feather shaft samples) from 4 LPMs in Yangzhou and in 178 human blood samples from healthy persons living in Yangzhou. The DNA from the feather shafts and human blood were extracted as previously described (8). PCR was performed by using the following 2 primers: AGV2_F 5′−CGTGTCCGCCAGCAGAAACGAC-′3 and AGV2_R 5′-GGTAGAAGCCAAAGCGTCCACGA-′3. The PCR targets partial VP2 and VP3 genes that have an expected size of 346 bp. The parameters of the PCR were as follows: 1 cycle at 95°C for 5 min; then 30 cycles at 94°C for 30 s, 64°C for 30 s, and 72°C for 30 s; and 1 cycle at 72°C for 10 min. PCR showed that a band with the size of ≈346 bp could be amplified in 10 of 54 chicken feather samples and in 2 of 178 human blood samples.

We confirmed the AGV2 specificity of these PCR–amplified bands by direct sequencing using the Sanger method. The sequence assay showed that the 12 sequences identified here had 98.3%–100% homology to each other and 92.2%–99.1% aa identity to AGV2 samples previously deposited in GenBank (see Figure legend for accession numbers). The positive rates for samples from the 4 LPMs tested were 25%, 12.5%, 15.8%, and 20%; the positive rate for the 178 human blood samples was 1.1%. The low positive frequency of AGV2 in human blood detected in this study is consistent with that found by investigation in other countries (3,4). Because the limit of detection of PCR in this study was estimated to be 2.7 copies of AGV2 DNA using dilutions of a plasmid with partial AGV2 sequence, we determined that the copy number of AGV2 in the 2 positive human blood samples was 2.7 × 103 copies/mL plasma.


Thumbnail of Phylogenetic analysis of AGV2. The phylogenetic tree was constructed by using the neighbor-joining method (1,000 bootstraps) with MEGA6 (9). Black squares indicate the 10 AGV2 identified from live poultry markets; black triangles indicate the 2 AGV2 identifed from human blood; black dot indicates the prototype AGV2 sequence. Sequences and GenBank accession nos.: AGV2–2014YZ01, KP993124; AGV2–2014YZ03, KP993125; AGV2–2014YZ04, KP993126; AGV2–2014YZ05, KP993127; AGV2–2014YZ07, KP99312

Figure. Phylogenetic analysis of AGV2. The phylogenetic tree was constructed by using the neighbor-joining method (1,000 bootstraps) with MEGA6 (9). Black squares indicate the 10 AGV2 identified from live poultry...

We also constructed a phylogenetic tree using the neighbor-joining method (1,000 bootstrap replications) with MEGA6 (9). The tree analysis revealed that the 12 AGV2 isolates we identified and 7 AGV2 isolates from GenBank clustered into 2 subgroups on the basis of the PCR amplified fragment (Figure). The 12 AGV2 sequences we identified clustered together with gyrovirus sequences detected in ferret and human samples in subgroup I, and the prototype sequence Ave3 was located in subgroup II. The 12 AGV2 showed ≈92.2%–93% aa identity to Ave3, and <99.1% homology with isolates CL33, G13, and 915F06007 detected in ferret and human samples. The 12 AGV2 sequences also showed ≈93%–93.9% identities to ACV2 sequence that was previously identified in human fecal samples from mainland China (GenBank accession no. JQ690763). The China sequence also clustered with Ave3 in subgroup II. These findings indicate that ≥2 subgroups of AGV2 are circulating in mainland China.

Our results demonstrate the presence of AGV2 in LPMs and human blood in mainland China. The amplification and analysis of partial AGV2 sequences was the major limitation in our method. The high homology between sequences identified in LPMs and human blood indicates the LPMs are a potential source for AGV2 in humans. Unlike our 12 conserved AGV2, AGV2 identified by Santos et al. in southern Brazil varied <15.8%, and these variants of AGV2 were mainly detected in diseased chickens (8). However, little is known about the molecular epidemiology of these AGV2 variants in other countries. More recently, Varela et al. reported the detection of AGV2 in poultry vaccines, indicating the potential role of contaminated vaccines in the spread of AGV2 (10). Future studies should investigate the large geographic distribution of AGV2 and monitor the variants, the host range, and the associated diseases.



This work was supported by the National Natural Science Foundation of China (31402228), National College Student Innovation Training Project (201411117002), Key University Science Research Project of Jiangsu Province (14KJA230002), Jiangsu Province College Student Innovation training Projects (201411117002Z and 201411117056Y), and the Priority Academic Program Development of Jiangsu Higher Education Institutions.


Jianqiang Ye1Comments to Author , Xiaoyan Tian1, Quan Xie, Yu Zhang, Yuanzhao Sheng, Zhenwen Zhang, Zhangping Yang, Hong Zhu, Yumeng Wang, Hongxia Shao, and Aijian Qin

Author affiliations: Ministry of Education Key Laboratory for Avian Preventive Medicine and Key Laboratory of Jiangsu Preventive Veterinary Medicine, Yangzhou University, Yangzhou, China (J. Ye, X. Tian, Q. Xie, Y. Zhang, Y. Sheng, H. Zhu, Y. Wang, H. Shao, A. Qin); Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou (J. Ye, Z. Zhang, C. Wang, H. Shao, A. Qin); College of Medicine, Yangzhou University, Yangzhou (Z. Zhang)



  1. Rijsewijk  FA, Dos Santos  HF, Teixeira  TF, Cibulski  SP, Varela  AP, Dezen  D, Discovery of a genome of a distant relative of chicken anemia virus reveals a new member of the genus Gyrovirus. Arch Virol. 2011;156:1097100 . DOIPubMed
  2. Sauvage  V, Cheval  J, Foulongne  V, Gouilh  MA, Pariente  K, Manuguerra  JC, Identification of the first human gyrovirus, a virus related to chicken anemia virus. J Virol. 2011;85:794850 . DOIPubMed
  3. Maggi  F, Macera  L, Focosi  D, Vatteroni  ML, Boggi  U, Antonelli  G, Human gyrovirus DNA in human blood, Italy. Emerg Infect Dis. 2012;18:9569. DOIPubMed
  4. Biagini  P, Bedarida  S, Touinssi  M, Galicher  V, de Micco  P. Human gyrovirus in healthy blood donors, France. Emerg Infect Dis. 2013;19:10145 . DOIPubMed
  5. Chu  DK, Poon  LL, Chiu  SS, Chan  KH, Ng  EM, Bauer  I, Characterization of a novel gyrovirus in human stool and chicken meat. J Clin Virol. 2012;55:20913 . DOIPubMed
  6. J Bullenkamp DC. Malik F, Alkhatabi H, Kulasekararaj A, Odell EW, Farzaneh F, Gäken J, et al. Human gyrovirus apoptin shows a similar subcellular distribution pattern and apoptosis induction as the chicken anaemia virus derived VP3/Apoptin. Cell Death Dis. 2012;3:e296.
  7. Abolnik  C, Wandrag  DB. Avian gyrovirus 2 and a virulent Newcastle disease virus coinfection in a chicken flock with neurologic symptoms and high mortalities. Avian Dis. 2014;58:904 . DOIPubMed
  8. dos Santos  HF, Knak  MB, de Castro  FL, Slongo  J, Ritterbusch  GA, Klein  TA, Variants of the recently discovered avian gyrovirus 2 are detected in southern Brazil and the Netherlands. Vet Microbiol. 2012;155:2306. DOIPubMed
  9. Tamura  K, Stecher  G, Peterson  D, Filipski  A, Kumar  S. MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol. 2013;30:27259. DOIPubMed
  10. Varela  AP, Dos Santos  HF, Cibulski  SP, Scheffer  CM, Schmidt  C, Sales Lima  FE, Chicken anemia virus and avian gyrovirus 2 as contaminants in poultry vaccines. Biologicals. 2014;42:34650. DOIPubMed




Cite This Article

DOI: 10.3201/eid2108.150203

1These authors contributed equally to this article.

Related Links


Table of Contents – Volume 21, Number 8—August 2015


Please use the form below to submit correspondence to the authors or contact them at the following address:

Jianqiang Ye, Yangzhou University, 12 Wenhui East Rd, Yangzhou, Jiangsu, 225009, China; or ,

Send To

character(s) remaining.

Comment submitted successfully, thank you for your feedback.


Page created: July 15, 2015
Page updated: July 15, 2015
Page reviewed: July 15, 2015
The conclusions, findings, and opinions expressed by authors contributing to this journal do not necessarily reflect the official position of the U.S. Department of Health and Human Services, the Public Health Service, the Centers for Disease Control and Prevention, or the authors' affiliated institutions. Use of trade names is for identification only and does not imply endorsement by any of the groups named above.