jueves, 5 de marzo de 2015

Ahead of Print -Bat Coronavirus in Brazil Related to Appalachian Ridge and Porcine Epidemic Diarrhea Viruses - Volume 21, Number 4—April 2015 - Emerging Infectious Disease journal - CDC

Ahead of Print -Bat Coronavirus in Brazil Related to Appalachian Ridge and Porcine Epidemic Diarrhea Viruses - Volume 21, Number 4—April 2015 - Emerging Infectious Disease journal - CDC



CDC. Centers for Disease Control and Prevention. CDC 24/7: Saving Lives. Protecting People.

Volume 21, Number 4—April 2015

Letter

Bat Coronavirus in Brazil Related to Appalachian Ridge and Porcine Epidemic Diarrhea Viruses

Technical Appendicies

Downloads

To the EditorTadarida brasiliensis (I. Geoffroy, 1824) is a species of free-tailed bat that has resident and migratory populations in Brazil (1). This species has adapted to urban areas, occupying roofs, ceilings, and other human constructions, and often coexists with other bat species and humans (2), enabling epidemiologic risks (3). In recent studies, an alphacoronavirus has been detected in urban bat species Molossus molossusM. rufus, andTadarida brasiliensi in Brazil (4,5). Evidence suggests that alphacoronaviruses may use bats as hosts to spread human coronavirus (HCoV) NL63, which originated by evolution of Appalachian Ridge CoV strain 2 (ARCoV.2) (6).
In this study, a total of 20 anal and tracheal swab samples from 10 bats (T. brasiliensis) were collected at the Jequitibás Wood, in Campinas, São Paulo State, Brazil (22°54′31.34′′S 47°02′58.01′′W). We extracted viral genetic material using the RNA Extraction Mini Kit (QIAGEN, Hilden, Germany) and synthesized cDNA using random primers from the High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA, USA), following the manufacturer’s protocol.
Samples were analyzed by conventional reverse transcription PCR assays using panCoV primers targeting a 215-bp replicase fragment as previously described (7) with slight modifications to include more cycles and less extension time in order to obtain more PCR products. Sequencing reactions on a Pancoronavirus-positive anal swab sample (7) were performed at Central Laboratory of High Performance Technologies in Life Sciences (LaCTAD) at UNICAMP (http://www.lactad.unicamp.br) using an automated sequencer (3730xl DNA Analyzer; Applied Biosystems).
The chromatograms were edited using the program UGENE version 1.14 (UGENE, http://ugene.unipro.ru/forum/YaBB.pl?num=1407749393) and evaluated using Phred scores for base calling. Alignment was made with ClustalW v.2.1 software (http://www.clustal.org) implemented on Linux command line interface, and a similarity matrix was generated with sequences retrieved from the GenBank database. A 144-nt fragment of the replicase gene was obtained after editing, and phylogenetic analysis was performed after determining the best evolution model by using the jModelTest2 software (https://code.google.com/p/jmodeltest2/). Different CoV sequences were included to represent the genera Alpha-, Beta-, Gamma-, andDeltacoronavirus. Clustering with the ARCoV.2 and porcine epidemic diarrhea virus (PEDV) was obtained using the maximum-likelihood (ML) method after 1,000 Shimodaira-Hasegawa–like support values with the general time-reversible model and category approximation in 20 rates category in a gamma distribution (Technical Appendix[PDF - 57 KB - 2 pages] Figure, panel A) and neighbor-joining methods under Kimura-2-parameter and 1,000 replicates of bootstrap (Technical Appendix[PDF - 57 KB - 2 pages] Figure, panel B).
Subsequently, metagenomic analysis was made by creating a pool of the 10 bat samples. Samples were resuspended in Dulbecco Modified Eagle Medium (Life Technologies-GIBCO, Grand Island, NY, USA) and filtered through 0.22 μm. The recovered sample was then treated with DNase (Invitrogen, Carlsbad, CA, USA) to remove contaminating DNA and with Proteinase K (Invitrogen) to eliminate inhibitors and to disrupt viral capsids. Samples were then subjected to RNA extraction (QIAGEN) and sent to the sequencing core facility. Sequencing was performed on Illumina HiSeq2500 instrument by using the 2×100 bp kit according to manufacturer’s instructions.
Through these analyses, we obtained 34,409,110 reads, of which 76.47% had quality index ≥30. The contigs were assembled by de novo genome assembly (blastx E-value ≤1–5) (8) generating 10.742 scaffolds: 35 matches for coronaviruses (using the Coronavirus Database, http://covdb.microbiology.hku.hk), 3 matches for PEDV, and 2 matches for HCoV-NL63 (both using the UniProt database, http://www.uniprot.org) (Technical Appendix[PDF - 57 KB - 2 pages]). The sequences obtained had 87.5% (126/144) nucleotide identity with ARCoV.2, an unclassified alphacoronavirus (GenBank accession no. JX537912) for which a zoonotic role has been suggested (6). Preliminary analysis indicated good coverage of the polymerase region of the ARCoV.2 reference sequence by the reads (quality index >40) by using reference assembly against CoV complete genomes. This finding reinforces the hypothesis of this viral agent in the specimens analyzed. Moreover, molecular assays are underway in our laboratory to elucidate the alternative hypothesis of PEDV presence in bats in Brazil.
In summary, we found that a CoV detected in T. brasiliensis bats in Brazil has close phylogenetic relationships to ARCoV.2 and PEDV. Considering the zoonotic impact of these viral agents on the emergence of new diseases in animal and human populations, we believe that both results may strongly contribute to a better understanding of the molecular eco-epidemiology of these alphacoronaviruses. The reconstruction of their evolutionary history to trace their occurrence in humans and in bat populations as well as in other animals is being conducted to clarify their evolutionary pathway.
Paulo Vitor Marques Simas1, Ana Caroline de Souza Barnabé1, Ricardo Durães-Carvalho1, Daniel Ferreira de Lima Neto1, Leonardo Cardia Caserta1, Luiza Artacho, Fábio André Facco Jacomassa, Matheus Cavalheiro Martini, Márcia Mercês Aparecida Bianchi dos Santos, Paulo Anselmo Nunes Felippe, Helena Lage Ferreira, and Clarice Weis ArnsComments to Author 
Author affiliations: State University of Campinas, Campinas, São Paulo, Brazil (P.V.M. Simas, A.C.S. Barnabé, R. Durães-Carvalho, D.F. Lima-Neto, L.C. Caserta, L. Artacho, M.C. Martini, P.A.N. Felippe)São Paulo State University, Rio Claro, São Paulo (F.A.F. Jacomassa)Federal University of Juiz de Fora, Juiz de Fora, Brazil (M.M.A. Bianchi dos Santos)São Paulo University, Pirassununga, Brazil (H.L. Ferreira).

Acknowledgment

This work was supported by FAPESP (grant 2011/50919-5) and CNPq (grant 307738/2011-6).

References

  1. Armstrong KBrazilian free-tailed bat (Tadarida brasiliensis). [cited 2014 Dec 12]Mamm Species2008;4:16http://www.cfr.msstate.edu/wildlife/mammals/pdf/Brazilianfree-tailedbat.pdf.
  2. Wilkins KT. Tadarida brasiliensis. Mamm Species. Oxford University Press. 1989;331:1–10.
  3. Scheffer KCCarrieri MLAlbas ASantos HCPKotait IIto FHRabies virus in naturally infected bats in the state of São Paulo, southeastern Brazil.Rev Saude Publica2007;41:38995 . DOIPubMed
  4. Góes LGBRuvalcaba SGCampos AAQueiroz LHDe Carvalho CJerez JANovel Bat Coronaviruses, Brazil and Mexico. Emerg Infect Dis.2013;19:17113 .PubMed
  5. Lima FECampos FSFilho HBatista HBJunior PCibulski SPDetection of Alphacoronavirus in velvety free-tailed bats (Molossus molossus) and Brazilian free-tailed bats (Tadarida brasiliensis) from urban areas of Southern Brazil. Virus Genes2013;47:1647 . DOIPubMed
  6. Huynh JLi SYount BSmith ASturges LOlsen JCEvidence Supporting a Zoonotic Origin of Human Coronavirus Strain NL63. J Virol.2012;86:1281625 . DOIPubMed
  7. Vijgen LMoes EKeyaerts ELi SVan Ranst MA pancoronavirus RT-PCR assay for detection of all known coronaviruses. Methods Mol Biol.2011;454:312DOIPubMed
  8. Peng YHenry CMLeung, S. M. Yiu, Francis Y. L. Chin. IDBA-UD: a de novo assembler for single-cell and metagenomic sequencing data with highly uneven depth. Bioinformatics2012;28:14208 . DOIPubMed

Technical Appendix

Suggested citation for this article: Simas PVM, Barnabé ACS, Durães-Carvalho R, de Lima Neto DF, Caserta LC, Artacho L, et al. Bat coronavirus in Brazil Related to Appalachian Ridge and Porcine Epidemic Diarrhea Viruses [letter]. Emerg Infect Dis. 2015 Apr [date cited].http://dx.doi.org/10.3201/eid2104.141783
DOI: 10.3201/eid2104.141783


1These authors contributed equally to this article.
Medline cannot find the journal "Mamm Species" (in reference 1 "Armstrong, 2008"). Please check the journal name.
CrossRef reports the year should be "2008" not "2011" in reference 7 "Vijgen, Moes, Keyaerts, Li, Van Ranst, 2011".
Medline reports the year should be "2008" not "2011" in reference 7 "Vijgen, Moes, Keyaerts, Li, Van Ranst, 2011".

Ahead of Print -Enterovirus A71 Subgenotype B5, France, 2013 - Volume 21, Number 4—April 2015 - Emerging Infectious Disease journal - CDC

Ahead of Print -Enterovirus A71 Subgenotype B5, France, 2013 - Volume 21, Number 4—April 2015 - Emerging Infectious Disease journal - CDC



CDC. Centers for Disease Control and Prevention. CDC 24/7: Saving Lives. Protecting People.

Volume 21, Number 4—April 2015

Letter

Enterovirus A71 Subgenotype B5, France, 2013

On This Page

Figures

Downloads

To the Editor: We report the detection of human enterovirus A71 (EV-A71) subgenotype B5 in France, 6 years after it was first detected in Europe. EV-A71 belongs to the Enterovirus A species (genus Enterovirus, familyPicornaviridae) and is a major cause of hand, foot and mouth disease (HFMD), sometimes associated with severe neurologic complications (1). EV-A71 strains are classified in 6 genotypes, A–F, (2) but most of the circulating strains belong to genotypes B and C and to 11 subgenotypes (B0−B5, C1−C5) (1). Genotypes B and C have been reported in HFMD epidemics in the Asia-Pacific region; different subgenotypes cause nationwide epidemics that usually occur every 2−3 years (1). During 1963−1986 in Europe, Australia, and the United States, enterovirus infections were caused by viruses from subgenotypes B0, B1, and B2, but since 2000, infections with C1 and C2 viruses have begun to predominate (3,4). The other subgenotypes have been reported rarely in Europe; the C4b and C4a strains were identified in France, Germany, Austria, and Denmark in 2004 and 2012, respectively (46), and the B5 subgenotype was reported in Denmark in 2007 (7).
In November 2013, a 3-week-old boy was admitted to the emergency unit of a hospital in Compiègne, France, with a 48-hour history of fever and irritability. He was born at term after an uneventful pregnancy and delivery. On admission, he had normal vital signs. He had had contact with a cousin with oral ulcerations, but no information was available about the source of the cousin’s infection.
Laboratory testing revealed moderate cytolytic hepatitis. Complete blood count results were within reference values. Cerebrospinal fluid showed pleiocytosis (38 leukocytes, 81% polymorphonuclear cells) with protein and glucose levels with reference ranges. Bacterial cultures of blood and cerebrospinal fluid were negative. Enterovirus genome was detected in serum samples and cerebrospinal fluid by reverse transcription PCR. The infant made a steady recovery and was discharged 10 days after admission, with no apparent adverse outcome. Final diagnosis was neonatal enterovirus infection with meningitis.
Genotyping was performed on the serum specimen by using seminested reverse transcription PCR amplification and sequencing of the viral protein 1 gene. Phylogenetic investigation with sequences of reference strains representing all subgenotypes indicated that the isolate from the patient, designated PAR024102_FRA13, belonged to the EV-A71 B5 subgenotype. We investigated the putative origin of the strain by comparing 248 nonredundant complete 1D sequences of EV-A71 B5 strains following a Bayesian phylogenetic approach. PAR024103_FRA13 shared a most recent common ancestor (posterior probability = 1) with virus strains sampled in China in 2009 and in Taiwan during the 2011–2012 outbreak, (8) but was only distantly related to them (data not shown).
Thumbnail of Phylogeny of enterovirus A71 (EV-A71) subgenogroups B4 and B5 inferred with 274 partial 1D gene sequences, France. Black diamond indicates strain PAR024103_FRA13 from this study. The phylogenetic relationships were inferred following a Bayesian method by using a relaxed molecular clock model with an uncorrelated exponential distribution of evolution rates estimated with a general time reversible substitution model and a Bayesian skyline plot as a population model (BEAST version 1.7.
Figure. Phylogeny of enterovirus A71 (EV-A71) subgenogroups B4 and B5 inferred with 274 partial 1D gene sequences, France. Black diamond indicates strain PAR024103_FRA13 from this study. The phylogenetic relationships were inferred following...
Further analyses with 274 partial 1D gene sequences from GenBank (on March 19, 2014) indicated close genetic relationships (posterior probability = 1) with strains isolated in Thailand in 2012 (9) (Figure). The complete genome of PAR024103_FRA13 was determined by nucleotide sequencing of 4 overlapping segments obtained by gene amplification (GenBank accession no. LK985324). Sequence comparisons were performed with 13 available EV-A71 B5 complete genomes; the virus strain isolated in France exhibited 92%−99.5% nt similarity (98.9%−99.4% aa similarity) throughout the genome.
The EV-A71 B5 subgenotype was first detected in 1999 in Malaysia and spread to several other countries in Asia during the 2000s. Outbreaks causing severe illness and deaths were reported in Japan (2003), Brunei (2006), and Taiwan (2008 and 2012) (1,8). The first detection of subgenotype B5 in Europe was associated with a recrudescence of EV-A71 infections associated with meningitis and HFMD in Denmark in 2007 (7). The overall phylogenetic data are consistent with an introduction of EV-A71 B5 in France by importation of a strain from Asia, possibly from Thailand. Transmission of EV-A71 strains has been shown to occur in Europe as discrete and temporally defined virus introductions, occasionally followed by sustained dissemination (C. Hassel, unpub. data).
The emergence of the Asiatic lineage EV-A71 C4a in Denmark in 2012 is a recent event (6). The reemergence of EV-A71 subgenotype B5 in 2008 in Taiwan resulted in the largest outbreak of EV-A71 infection in the past 11 years (8). To our knowledge, the B5 subgenotype has not previously been detected in Europe. Global herd immunity produced by circulation of the C2 genotype may protect the European population from the spread of other subgenotypes (3). However, given that most countries in Europe do not perform specific surveillance for HFMD and most enterovirus infections are asymptomatic, this particular subgenotype could be circulating more widely without detection.
Enterovirus infections in neonates and infants are a frequent cause of hospitalization, which may contribute to EV-A71 detection (5). However, the development of a national syndromic surveillance targeting HFMD would enable early detection of HFMD outbreaks and any new EV-A71 subgenotype. Attention should also be paid to the potential risks of epidemic spread of EV-A71 outside Asia posed by international travelers.
Audrey MirandComments to Author , Lucie Molet, Chervin Hassel, Hélène Peigue-Lafeuille, Flore Rozenberg, Jean-Luc Bailly, and Cécile Henquell
Author affiliations: Clermont Université, Clermont-Ferrand, France (A. Mirand, C. Hassel, H. Peigue-Lafeuille, J.-L. Bailly, C. Henquell)Centre National de Référence des Entérovirus-Parechovirus, Clermont-Ferrand (A. Mirand, H. Peigue-Lafeuille, J.-L. Bailly, C. Henquell)Hôpital Cochin, Paris, France (L. Molet, F. Rozenberg)Université Paris Descartes, Paris (L. Molet, F. Rozenberg)

Acknowledgments

We are grateful to Nathalie Rodde and Gwendoline Jugie for excellent technical assistance in enterovirus genotyping and complete genome sequencing. We thank Jeffrey Watts for revising the English manuscript.
The Centre National de Référence des Enterovirus-Parechovirus is supported by an annual grant from the French national public health network (Institut de Veille Sanitaire).

References

  1. Solomon TLewthwaite PPerera DCardosa MJMcMinn POoi MHVirology, epidemiology, pathogenesis, and control of enterovirus 71. Lancet Infect Dis2010;10:77890DOIPubMed
  2. Bessaud MRazadindratsimandresy RNougairède AJoffret MLDeshpande JMDubot-Pérès AMolecular comparison and evolutionary analyses of VP1 nucleotide sequences of new African human enterovirus 71 isolates reveal a wide genetic diversity. PLoS One2014;9:e90624.DOIPubMed
  3. van der Sanden Svan der Avoort, Lemey P, Uslu G, Koopmans M. Evolutionary trajectory of the VP1 gene of human enterovirus 71 genogroup B and C viruses. J Gen Virol2010;91:194958DOIPubMed
  4. Mirand ASchuffenecker IHenquell CBillaud GJugie GFalcon DPhylogenetic evidence of a recent spread of two populations of human enterovirus 71 in European countries. J Gen Virol2010;91:226377DOIPubMed
  5. Schuffenecker IHenquell CMirand ACoste-Burel MMarque-Juillet SDesbois DNew introductions of enterovirus 71 subgenogroup C4, France, 2012. Emerg Infect Dis. 2014;20:1343–6.
  6. Fischer TKNielsen AYSydenham TVAndersen PHAndersen BMidgley SEEmergence of enterovirus 71 C4a in Denmark, 2009 to 2013. Euro Surveill2014;19;20911.PubMed
  7. Badran SAMidgley SAndersen PBöttinger BClinical and virological features of enterovirus 71 infections in Denmark, 2005 to 2008. Scand J Infect Dis2011;43:6428DOIPubMed
  8. Wu WHKuo TCLin YTHuang SWLiu HF, Wang J, et al. Molecular epidemiology of enterovirus 71 infection in the central region of Taiwan from 2002 to 2012. PLoS One2013;8:e83711DOIPubMed
  9. Linsuwanon PPuenpa JHuang SWWang YFMauleekoonphairoj JWang JREpidemiology and seroepidemiology of human enterovirus 71 among Thai populations. J Biomed Sci2014;21:16DOIPubMed

Figure



DOI: 10.3201/eid2104.141093

Ahead of Print -Enterovirus D68 Infection, Chile, Spring 2014 - Volume 21, Number 4—April 2015 - Emerging Infectious Disease journal - CDC

Ahead of Print -Enterovirus D68 Infection, Chile, Spring 2014 - Volume 21, Number 4—April 2015 - Emerging Infectious Disease journal - CDC



CDC. Centers for Disease Control and Prevention. CDC 24/7: Saving Lives. Protecting People.



Volume 21, Number 4—April 2015

Letter

Enterovirus D68 Infection, Chile, Spring 2014

Tables

Downloads

To the Editor: Enterovirus D68 (EV-D68) is an emergent viral pathogen associated with severe respiratory illness, especially in children with asthma (1). The ongoing epidemic in the United States has expanded to 47 states; as of November 25, 2014, a total of 1,121 persons were affected (http://www.cdc.gov/non-polio-enterovirus/outbreaks/EV-D68-outbreaks.html).
In Chile, because of clinical suspicion of infection caused by the same EV-D68 reported in the United States, we conducted further testing. We selected 6 children who were at Clinica Las Condes (CLC), Santiago, during September–October 2014, for whom nasopharyngeal samples were enterovirus positive according to multiplex PCR (CLART PneumoVir; Genomica, Madrid, Spain) or rhinovirus/enterovirus positive according to the FilmArray Respiratory Panel (BioFire Diagnostics, Salt Lake City, UT, USA). The CLC ethics committee authorized the study. CLC is the second largest private hospital in eastern Santiago, which has a population of nearly 400,000. Since 2008, detection of respiratory viruses by CLART PneumoVir has been performed for all hospitalized CLC patients with respiratory disease. This test detects 17 respiratory viruses, including enterovirus (generic detection). Since 2014, CLC also incorporated testing with the FilmArray Respiratory Panel, which does not distinguish enterovirus from rhinovirus.
We sent 6 nasopharyngeal samples from the children to the Centers for Disease Control and Prevention for detection of EV-D68 by real-time reverse transcription PCR (http://www.cdc.gov/non-polio-enterovirus/downloads/ev-d68-rt-pcr-protocol.pdf) and sequencing (2). Of the 6 patients, EV-D68 was confirmed for 2 patients.
Patient 1 was a boy, 7.5 years of age, who was hospitalized on September 21, 2014, with a history of asthma since 3 years of age. During the previous 3 months, his father had frequently traveled to the United States. The patient’s current episode began with upper respiratory symptoms and low-grade fever (38°C) and was followed by intense vomiting. On admission, the child exhibited respiratory distress and an oxygen saturation of 88%, which led to his admission to the pediatric intensive care unit and management with noninvasive mechanical ventilation (bilevel positive airway pressure) for 48 hours. He was discharged home after 5 days of hospitalization, having required supplemental oxygen for 4 of those days.
Patient 2 was a 9-year-old boy who was hospitalized on October 1, 2014, with a history of severe asthma since 7 years of age. He had visited the emergency department multiple times for asthma crises. Both parents and his 2 brothers also had asthma. The episode reported here began 4 days before hospitalization, with a dry cough and progressive breathing difficulty, requiring 3 emergency department visits. During the third visit, low oxygen saturation (91%) led to hospitalization. The child had no fever during this entire episode. He received up to 50% fraction of inspired oxygen, intravenous methylprednisolone, intravenous magnesium sulfate, and inhaled salbutamol; he was discharged in good medical condition after 8 days of hospitalization, having received supplemental oxygen for 7 of those days.
These 2 EV-D68–positive patients had marked pulmonary hyperinsufflation and required prolonged oxygen therapy. Sequence analysis of the viral protein 1 gene revealed that both of these viruses clustered with the major outbreak strain from the United States. Partial gene sequences of viral protein 1 were deposited in the GenBank database under accession nos. KP247599 and KP247600.
We next used CLART PneumoVir to retest samples that had been positive for enterovirus/rhinovirus by FilmArray during September–October 2013 and 2014. The number of overall samples tested for respiratory viruses did not increase from 2013 to 2014 (227 and 218, respectively), but the percentage of enteroviruses detected increased strikingly (from 2.6% to 14.6%). We then compared clinical characteristics and their frequency of occurrence among enterovirus-positive patients hospitalized during September–October 2013 and September–October 2014. The clinical features of 24 enterovirus-positive patients hospitalized during 2014 differed from those of 5 enterovirus-positive children hospitalized during 2013. Hospitalization in 2014 was mostly for asthmatic crisis in children 2.5 to 7 years of age; this pattern is less clear for the few patients hospitalized in 2013 (Table). A substantial proportion of patients hospitalized in 2014 required oxygen support and admission to the pediatric intensive care unit.
In conclusion, we report 2 confirmed cases of EV-D68 in a Southern Hemisphere country during the 2014 outbreak reported in the United States. That these cases are virologically and clinically related to those reported in the United States documents that the virus had been introduced to the Southern Hemisphere during the spring of 2014. A substantial increase in enterovirus cases displaying a notably similar clinical pattern (asthmatic crisis in children) strongly suggests that EV-D68 infections are increasingly rapidly. This virus has been previously identified in the region (3) but only sporadically. The virus could spread to other areas in Santiago and to other cities, and similar situations could occur in other Latin American countries, especially those with many residents who travel to the United States. Public health officials need to be notified of this potential, and appropriate surveillance and treatment strategies need to be implemented.
Juan P. TorresComments to Author , Mauricio J. Farfan, Giannina Izquierdo, Paula Piemonte, Juan Henriquez, and Miguel L. O’Ryan
Author affiliations: Clinica Las Condes, Santiago, Chile (J.P. Torres, M.J. Farfan, G. Izquierdo, P. Piemonte, J. Henriquez, M.L. O’Ryan)University of Chile, Santiago (J.P. Torres, M.J. Farfan, G. Izquierdo, M.L. O’Ryan)

Acknowledgment

We are indebted to and pleased to acknowledge W. Allan Nix and his team for their analysis of the samples from Chile; we also thank Grupo Bios Chile for their valuable support and for providing a CLART PneumoVir kit.

References

  1. Midgley CMJackson MASelvarangan RTurabelidze GObringer EJohnson DSevere respiratory illness associated with enterovirus D68—Missouri and Illinois, 2014. MMWR Morb Mortal Wkly Rep2014;63:7989 .PubMed
  2. Nix WAOberste MSPallansch MASensitive, seminested PCR amplification of VP1 sequences for direct identification of all enterovirus serotypes from original clinical specimens. J Clin Microbiol2006;44:2698704DOIPubMed
  3. Garcia JEspejo VNelson MSovero MVillaran MVGomez JHuman rhinoviruses and enteroviruses in influenza-like illness in Latin America. Virol J2013;10:305DOIPubMed

Table

Suggested citation for this article: Torres JP, Farfan MJ, Izquierdo G, Piemonte P, Henriquez J, O’Ryan ML. Enterovirus D68 infection, Chile, spring 2014 [letter]. Emerg Infect Dis. 2015 Apr [date cited]. http://dx.doi.org/10.3201/eid2104.141766
DOI: 10.3201/eid2104.141766


Related Links

Ahead of Print -Human Parvovirus 4 Infection among Mothers and Children in South Africa - Volume 21, Number 4—April 2015 - Emerging Infectious Disease journal - CDC

Ahead of Print -Human Parvovirus 4 Infection among Mothers and Children in South Africa - Volume 21, Number 4—April 2015 - Emerging Infectious Disease journal - CDC



CDC. Centers for Disease Control and Prevention. CDC 24/7: Saving Lives. Protecting People.

Volume 21, Number 4—April 2015

Letter

Human Parvovirus 4 Infection among Mothers and Children in South Africa

Figures

Technical Appendicies

Downloads

To the Editor: Human parvovirus 4 (PARV4) is a single-stranded DNA virus in the family Parvoviridae (1). In Western countries, IgG against PARV4 is largely found only in persons with risk factors for parenteral infection and is strongly associated with co-infection with bloodborne viruses (24). In Africa, transmission seems to be more complicated; reported PARV4 seroprevalence is 4%–37%, even among persons at low risk and with no evidence of HIV or hepatitis C virus (HCV) co-infection (1,5,6).
The clinical significance of PARV4 infection remains uncertain. Infections may be asymptomatic, but a variety of clinical associations have been reported (1,7), including an increased rate of progression to AIDS in persons co-infected with HIV (8). This association raises particular concerns for many African populations in which these viruses are co-endemic.
To characterize the epidemiology of PARV4 infection in South Africa, we studied adults and children from pediatric outpatient clinics in Kimberley, South Africa, during May 2009–August 2013. Of the 157 participants, 90 were HIV-1–infected children, 24 their HIV-negative siblings, and 43 HIV-1–infected mothers (of whom 4 had >1 child enrolled). Approval was given by the Ethics Committee of the Faculty of Health Science, University of Free State, South Africa. Written consent was given by all adults and parents/guardians on behalf of their children.
Blood samples were collected from participants, and serum was tested for evidence of PARV4 infection by using ELISA (in duplicate) to detect IgG against PARV4 viral protein 2 (3,6) and by using PCR to detect PARV4 DNA (9). For 92 patients, HIV RNA loads were available; testing was performed by using the Abbott Laboratories m2000 platform (Abbott Park, IL, USA). For 118 of the HIV-infected patients, CD4+ T-cell counts were ascertained by flow cytometry. Statistical analyses were undertaken by using Prism version 6.0f and online software (http://graphpad.com/quickcalcs/). Confidence intervals were calculated by using the adjusted Wald method (http://www.measuringusability.com/wald.htm).
We detected IgG against PARV4 in 58 (37%) of 157 patients; this proportion is broadly comparable with that reported from other settings in sub-Saharan Africa, including Burkina Faso, the Democratic Republic of the Congo, and a previous cohort of HIV-infected persons in South Africa (5). Although routes of transmission in Africa remain to be characterized, these high seroprevalence rates support the possibility that some PARV4 transmission may be occurring by nonparenteral routes, as suggested by others (5,10).
Thumbnail of Relationship between age and seroprevalence of IgG against human parvovirus 4 (PARV4) among 157 mothers and children in Kimberley, South Africa, 2009–2013. A) Number and proportion of children and adults seropositive for IgG against PARV4; the number in each group is shown above the bar. p value calculated by using the Fisher exact test. B) Proportion of population seropositive for IgG against PARV4 according to age; the number in each group is shown above the bar. Data are shown fo
Figure. Relationship between age and seroprevalence of IgG against human parvovirus 4 (PARV4) among 157 mothers and children in Kimberley, South Africa, 2009–2013. A) Number and proportion of children and adults seropositive...
PARV4 IgG seroprevalence was higher among adults (49%) than children (33%), although this difference did not reach statistical significance (p = 0.07, Fisher exact test; Figure, panel A). We found a significant relationship between increasing age and PARV4 IgG serostatus (R2 = 0.59 by linear regression, p = 0.025; Figure, panel B). The numbers in each group are small, and further work is needed to define this association with more confidence. We did not detect any cases of PARV4 viremia, suggesting that chronic viremia or reactivation are probably uncommon, even among HIV-infected patients.
On the basis of previously reported data demonstrating PARV4 viremia in neonates (7), we hypothesized that vertical transmission is possible. To investigate further, we sought evidence of concordance between IgG serostatus of mothers and their children. Maternal PARV4 IgG status did not differ between IgG-positive and IgG-negative children (p = 1.00, Fisher exact test; Technical Appendix[PDF - 20 KB - 1 page] Table 1). The absence of correlation between the IgG statuses of mothers and children suggests that vertical transmission is probably not a major contributor to new infections, although it remains plausible that it may sometimes occur.
Data from Europe that suggest an association between PARV4 infection and progression to Centers for Disease Control and Prevention B-syndromes in HIV-positive persons are problematic because of confounding high rates of HCV infection and injection drug use in the PARV4-positive group (8). We sought evidence for this effect in our cohort, in which rates of HCV infection and injection drug use were likely to be negligible. We found no evidence of a PARV4 serostatus effect on HIV RNA load or CD4+ T cells in children (p = 0.13, p = 0.68, respectively; Technical Appendix[PDF - 20 KB - 1 page] Table 1) or adults (p = 0.15, p = 0.77, respectively;Technical Appendix[PDF - 20 KB - 1 page] Table 2).
We found an unexpected negative correlation between PARV4 IgG and HIV status in children (p = 0.05, Fisher exact test; Technical Appendix[PDF - 20 KB - 1 page] Table 1). One possible explanation is that a detectable PARV4 IgG response is not mounted or maintained in the context of HIV infection; however, this theory is not supported by previous studies in which PARV4 IgG seems to be more prevalent in HIV-infected populations (5,8).
Our analysis was limited by small numbers tested and the retrospective approach to sample testing. Demographic data were not recorded for this cohort, so we are unable to explore further possible social or demographic risk factors that might correlate with PARV4 infection.
This study contributes to an evolving body of data suggesting that PARV4 is highly endemic to different settings across Africa. The unknown clinical effects and transmission routes of this virus remain pressing questions for future research.
Philippa C. MatthewsComments to Author , Colin P. Sharp, Amna Malik, William F. Gregory, Emily Adland, Pieter Jooste, Philip J. R. Goulder, Peter Simmonds, and Paul Klenerman
Author affiliations: University of Oxford, Oxford, UK (P.C. Matthews, A. Malik, E. Adland, P.J.R. Goulder, P. Klenerman)Oxford University Hospitals, Oxford (P.C. Matthews, P. Klenerman)The University of Edinburgh, Midlothian, Scotland, UK (C.P. Sharp, W.F. Gregory, P. Simmonds)University of Free State, Kimberley, South Africa (P. Jooste)John Radcliffe Hospital, Oxford (P. Klenerman)

Acknowledgment

P.C.M. is a National Institute for Health Research Clinical Lecturer and received funding from Oxford University Clinical Academic Graduate School to cover the cost of this work. C.P.S., W.F.G., and P.S. were funded by the Biotechnology and Biological Sciences Research Council. P.J.R.G. is a Wellcome Trust Senior Investigator. P.K. is funded by the National Institute for Health Research Biomedical Research Centre, Oxford Martin Centre, and Wellcome Trust grant no. 091663.

References

  1. Matthews PCMalik ASimmons RSharp CSimmonds PKlenerman PPARV4: an emerging tetraparvovirus. PLoS Pathog2014;10:e1004036.DOIPubMed
  2. Sharp CPLail ADonfield SGomperts EDSimmonds PVirologic and clinical features of primary infection with human parvovirus 4 in subjects with hemophilia: frequent transmission by virally inactivated clotting factor concentrates. Transfusion2012;52:14829 . DOIPubMed
  3. Sharp CPLail ADonfield SSimmons RLeen CKlenerman PHigh frequencies of exposure to the novel human parvovirus PARV4 in hemophiliacs and injection drug users, as detected by a serological assay for PARV4 antibodies. J Infect Dis2009;200:111925DOIPubMed
  4. Simmonds PManning AKenneil RCarnie FWBell JEParenteral transmission of the novel human parvovirus PARV4. Emerg Infect Dis.2007;13:13868DOIPubMed
  5. Sharp CPVermeulen MNebie YDjoko CFLeBreton MTamoufe UEpidemiology of human parvovirus 4 infection in sub-Saharan Africa. Emerg Infect Dis2010;16:16057DOIPubMed
  6. Lavoie MSharp CPPepin JPennington CFoupouapouognigni YPybus OGHuman parvovirus 4 infection, Cameroon. Emerg Infect Dis.2012;18:6803DOIPubMed
  7. Chen MYYang SJHung CCPlacental transmission of human parvovirus 4 in newborns with hydrops, Taiwan. Emerg Infect Dis2011;17:19546.DOIPubMed
  8. Simmons RSharp CMcClure CPRohrbach JKovari HFrangou EParvovirus 4 infection and clinical outcome in high-risk populations. J Infect Dis.2012;205:181620DOIPubMed
  9. Sharp CPLeBreton MKantola KNana ADiffo Jle DDjoko CFWidespread infection with homologues of human parvoviruses B19, PARV4, and human bocavirus of chimpanzees and gorillas in the wild. J Virol2010;84:1028996DOIPubMed
  10. May JDrexler JFReber USarpong NAdjei OPanning MHuman parvovirus 4 viremia in young children, Ghana. Emerg Infect Dis.2012;18:16902DOIPubMed

Figure

Technical Appendix

Suggested citation for this article: Matthews PC, Sharp CP, Malik A, Gregory WF, Adland E, Jooste P, et al. Human parvovirus 4 infection among mothers and children in South Africa. [letter]. Emerg Infect Dis. 2015 Apr [date cited]. http://dx.doi.org/10.3201/eid2104.141545


DOI: 10.3201/eid2104.141545