title: Molecular epidemiology of human enterovirus 71 at the origin of an epidemic of fatal hand, foot and mouth disease cases in Cambodia.
abstract: Human enterovirus 71 (EV-A71) causes hand, foot and mouth disease (HFMD). EV-A71 circulates in many countries and has caused large epidemics, especially in the Asia-Pacific region, since 1997. In April 2012, an undiagnosed fatal disease with neurological involvement and respiratory distress occurred in young children admitted to the Kantha Bopha Children's Hospital in Phnom Penh, Cambodia. Most died within a day of hospital admission, causing public panic and international concern. In this study, we describe the enterovirus (EV) genotypes that were isolated during the outbreak in 2012 and the following year. From June 2012 to November 2013, 312 specimens were collected from hospitalized and ambulatory patients and tested by generic EV and specific EV-A71 reverse transcription PCR. EV-A71 was detected in 208 clinical specimens while other EVs were found in 32 patients. The VP1 gene and/or the complete genome were generated. Our phylogenetic sequencing analysis demonstrated that 80 EV-A71 strains belonged to the C4a subgenotype and 3 EV-A71 strains belonged to the B5 genotype. Furthermore, some lineages of EV-A71 were found to have appeared in Cambodia following separate introductions from neighboring countries. Nineteen EV A (CV-A6 and CV-A16), 9 EV B (EV-B83, CV-B3, CV-B2, CV-A9, E-31, E-2 and EV-B80) and 4 EV C (EV-C116, EV-C96, CV-A20 and Vaccine-related PV-3) strains were also detected. We found no molecular markers of disease severity. We report here that EV-A71 genotype C4 was the main etiological agent of a large outbreak of HFMD and particularly of severe forms associated with central nervous system infections. The role played by other EVs in the epidemic could not be clearly established.
authors.affiliation: Pasteur Institute in Cambodia, Phnom Penh 12000, Cambodia.
authors.forename: Veasna
authors.initials: V
authors.lastname: Duong
authors.affiliation: Pasteur Institute in Cambodia, Phnom Penh 12000, Cambodia.
authors.forename: Channa
authors.initials: C
authors.lastname: Mey
authors.affiliation: Pasteur Institute, Paris 75724, France.
authors.forename: Marc
authors.initials: M
authors.lastname: Eloit
authors.affiliation: State Key Laboratory of Emerging Infectious Diseases, School of Public Health, The University of Hong Kong, Hong Kong, China.
authors.forename: Huachen
authors.initials: H
authors.lastname: Zhu
authors.affiliation: Pasteur Institute in Cambodia, Phnom Penh 12000, Cambodia.
authors.forename: Lucie
authors.initials: L
authors.lastname: Danet
authors.affiliation: Institut Pasteur in Shanghai, Shanghai 200025, China.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Huang
authors.affiliation: Institut Pasteur in Shanghai, Shanghai 200025, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Zou
authors.affiliation: Pasteur Institute in Cambodia, Phnom Penh 12000, Cambodia.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Tarantola
authors.affiliation: PathoQuest, Paris 75015, France.
authors.forename: Justine
authors.initials: J
authors.lastname: Cheval
authors.affiliation: Pasteur Institute, Paris 75724, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Perot
authors.affiliation: Kantha Bopha Hospital, Phnom Penh 12000, Cambodia.
authors.forename: Denis
authors.initials: D
authors.lastname: Laurent
authors.affiliation: Kantha Bopha Hospital, Phnom Penh 12000, Cambodia.
authors.forename: Beat
authors.initials: B
authors.lastname: Richner
authors.affiliation: Kantha Bopha Hospital, Phnom Penh 12000, Cambodia.
authors.forename: Santy
authors.initials: S
authors.lastname: Ky
authors.affiliation: Kantha Bopha Hospital, Phnom Penh 12000, Cambodia.
authors.forename: Sothy
authors.identifier: http://orcid.org/0000-0003-4681-6681
authors.initials: S
authors.lastname: Heng
authors.affiliation: Ministry of Health, Phnom Penh 12000, Cambodia.
authors.forename: Sok
authors.initials: S
authors.lastname: Touch
authors.affiliation: Ministry of Health, Phnom Penh 12000, Cambodia.
authors.forename: Ly
authors.initials: L
authors.lastname: Sovann
authors.affiliation: Oxford University Clinical Research Unit, Hospital for Tropical Diseases, Ho Chi Minh P1Q5, Vietnam.
authors.forename: Rogier
authors.identifier: http://orcid.org/0000-0002-9807-1821
authors.initials: R
authors.lastname: van Doorn
authors.affiliation: Oxford University Clinical Research Unit, Hospital for Tropical Diseases, Ho Chi Minh P1Q5, Vietnam.
authors.forename: Thanh
authors.initials: T
authors.lastname: Tan Tran
authors.affiliation: Oxford University Clinical Research Unit, Hospital for Tropical Diseases, Ho Chi Minh P1Q5, Vietnam.
authors.forename: Jeremy J
authors.initials: JJ
authors.lastname: Farrar
authors.affiliation: J. Craig Venter Institute, Rockville, MD 92037, USA.
authors.forename: David E
authors.initials: DE
authors.lastname: Wentworth
authors.affiliation: J. Craig Venter Institute, Rockville, MD 92037, USA.
authors.forename: Suman R
authors.identifier: http://orcid.org/0000-0003-2496-9724
authors.initials: SR
authors.lastname: Das
authors.affiliation: J. Craig Venter Institute, Rockville, MD 92037, USA.
authors.forename: Timothy B
authors.initials: TB
authors.lastname: Stockwell
authors.affiliation: Pasteur Institute, Paris 75724, France.
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Manuguerra
authors.affiliation: Pasteur Institute, Paris 75724, France.|National Institute for Health and Medical Research, INSERM U994, Paris 75000, France.
authors.forename: Francis
authors.initials: F
authors.lastname: Delpeyroux
authors.affiliation: State Key Laboratory of Emerging Infectious Diseases, School of Public Health, The University of Hong Kong, Hong Kong, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Guan
authors.affiliation: Institut Pasteur in Shanghai, Shanghai 200025, China.
authors.forename: Ralf
authors.initials: R
authors.lastname: Altmeyer
authors.affiliation: Pasteur Institute in Cambodia, Phnom Penh 12000, Cambodia.|GlaxoSmithKline Vaccines R&D, Singapore 189720, Singapore.
authors.forename: Philippe
authors.initials: P
authors.lastname: Buchy
mesh_terms: D000293:Adolescent; D000328:Adult; D002160:Cambodia / Q000453:epidemiology; D002648:Child; D002675:Child, Preschool; D004196:Disease Outbreaks; D029821:Enterovirus A, Human / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D058872:Epidemics; D005260:Female; D016679:Genome, Viral; D005838:Genotype; D006232:Hand, Foot and Mouth Disease / Q000453:epidemiology* / Q000401:mortality* / Q000821:virology; D006760:Hospitalization; D006801:Humans; D007223:Infant; D008297:Male; D010802:Phylogeny; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D055815:Young Adult
chemical_list: D012367:RNA, Viral