The number of cases and severity of disease associated with dengue infection in Sri Lanka has been increasing since 1989, when the first epidemic of dengue hemorrhagic fever was recorded. We identified a new dengue virus 1 strain circulating in Sri Lanka that coincided with the 2009 dengue epidemic

Abeysinghe, M.R. Nihal

Amarasinghe, Ananda

de Silva, Aravinda M.

De Silva, Aruna Dharshan

Gubler, Duane J.

Gunasena, Sunethra

Letson, G. William

Logendra, Barathy

Margolis, Harold S.

Ooi, Eng Eong

Palihawadana, Paba

Tam, Clarence C.

Tan, Ying

Tissera, Hasitha A.

Wahala, Wahala M.P.B.

Emerging Infectious Diseases

PubMed

Hasitha A. Tissera

Eng Eong Ooi

Duane J. Gubler

Ying Tan

Barathy Logendra

Wahala M.P.B. Wahala

Aravinda M. de Silva

M.R. Nihal Abeysinghe

Paba Palihawadana

Sunethra Gunasena

Clarence C. Tam

Ananda Amarasinghe

G. William Letson

Harold S. Margolis

Aruna Dharshan De Silva

Directory of Open Access Journals

New Dengue Virus Type 1 Genotype in Colombo, Sri Lanka

10.3201/eid1711.101893Emerging Infectious Diseases17112053-2055EIDI

Tissera, H.A.

Ooi, E.E.

Gubler, D.J.

Tan, Y.

Logendra, B.

Wahala, W.M.P.B.

de Silva, A.M.

Abeysinghe, M.R.N.

Palihawadana, P.

Gunasena, S.

Tam, C.C.

Amarasinghe, A.

Letson, G.W.

Margolis, H.S.

de Silva, A.D.

ScholarBank@NUS

New dengue virus type 1 genotype in Colombo, Sri Lanka

Hasitha Tissera

Eng Ooi

Duane Gubler

Wahala Wahala

Aravinda de Silva

Clarence Tam

Harold Margolis

Aruna De Silva

Crossref

Tissera, Hasitha

Ooi, Eng

Gubler, Duane

Wahala, Wahala

de Silva, Aravinda

Tam, Clarence

Margolis, Harold

De Silva, Aruna

Carolina Digital Repository

New Dengue Virus Type 1 Genotype in Colombo, Sri LankaHasitha A. Tissera, Eng Eong Ooi, Duane J. Gubler, Ying Tan, Barathy Logendra, Wahala M.P.B. Wahala, Aravinda M. de Silva, M.R. Nihal Abeysinghe, Paba Palihawadana, Sunethra Gunasena, Clarence C. Tam, Ananda Amarasinghe, G. William Letson, Harold S. Margolis, and Aruna Dharshan De SilvaThe number of cases and severity of disease associated with dengue infection in Sri Lanka has been increasing since 1989, when the fi rst epidemic of dengue hemorrhagic fever was recorded. We identifi ed a new dengue virus 1 strain circulating in Sri Lanka that coincided with the 2009 dengue epidemic.Dengue virus (DENV) is a fl avivirus transmitted by Aedes spp. mosquitoes. There are 4 distinct DENV serotypes (DENV-1–4). Infection with a single serotype leads to long-term protective immunity against the homologous serotype but not against other serotypes (1). Globally, dengue is an emerging disease that causes an estimated 50–100 million infections, 500,000 dengue hemorrhagic fever (DHF) cases, and 22,000 deaths annually (2,3).Epidemiologic and other studies indicate that risk factors for severe dengue include secondary infection with a heterologous serotype, the strain of infecting virus, and age and genetic background of the host. Studies are under way to further explore the role of these factors in severe disease (1,4).In Sri Lanka, serologically confi rmed dengue was fi rst reported in 1962 (5), but although all 4 virus serotypes were present and there were cases of DHF, only since 1989 has DHF been considered endemic to Sri Lanka (5). Dengue was made a reportable disease in Sri Lanka in 1996, and the largest epidemic (35,008 reported cases, 170 cases/100,000 population, and 346 deaths) occurred in 2009 (6). DHF epidemics in 1989 and 2002–2004 were associated with emergence of new clades of DENV-3 (7,8). We report a new DENV-1 genotype introduced to Sri Lanka before the 2009 epidemic.The StudyThe study was approved by the Ethical Review Committee of the Faculty of Medicine, University of Colombo, Sri Lanka, and the Institutional Research Board of the International Vaccine Institute, Seoul, South Korea. Serum samples were obtained in 2009 and early 2010 from patients as part of a Pediatric Dengue Vaccine Initiative (PDVI) fever surveillance study in Colombo, Sri Lanka. Samples were originally tested for dengue by reverse transcription PCR at Genetech Research Institute (Colombo, Sri Lanka). A random subset of dengue-positive samples of all 4 serotypes was sent to the Program in Emerging Infectious Diseases Laboratory at Duke–National University of Singapore Graduate Medical School, Singapore, for virus isolation and sequencing.RNA was extracted from virus isolates, subjected to standard reverse transcription PCR to confi rm the presence  Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 17, No. 11, November 2011 2053Author affi liations: Ministry of Health, Colombo, Sri Lanka (H.A. Tissera, M.R.N. Abeysinghe, P. Palihawadana); London School of Hygiene and Tropical Medicine, London, UK (H.A. Tissera, C.C. Tam); Duke–National University of Singapore Graduate Medical School, Singapore (E.E. Ooi, D.J. Gubler, Y. Tan); Genetech Research Institute, Colombo (B. Logendra, A.D. De Silva); University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA (W.M.P.B. Wahala, A.M. de Silva); Medical Research Institute, Colombo (S. Gunasena); and International Vaccine Institute, Seoul, South Korea (A. Amerasinghe, G.W. Letson, H.S. Margolis)DOI: http://dx.doi.org/10.3201/eid1711.101893Figure. Phylogenetic tree of dengue virus 1 (DENV-1) serotype viruses from Sri Lanka (SL), 2009–2010, and other DENV-1 viruses. The tree is based on a 498-bp (nt 2056–2554) fragment that encodes portions of the envelope protein and nonstructural protein 1. Phylogenetic analysis was conducted by using MEGA5 (10). Percentages of replicate trees in which the associated taxa clustered in the bootstrap test (1,000 replicates) are shown next to the branches. Genotype I (Asia) includes SL isolates from 2009–2010; genotype III (South Pacifi c) includes SL isolates from the early 1980s; genotype IV (Africa/Americas) includes SL isolates from the 1990s and early 2000s. Classifi cation and naming of DENV-1 genotypes are based on the report by Rico-Hesse (11). DV, dengue virus. Scale bar indicates nucleotide substitutions per site.of dengue virus, and serotyped as described (7). Samples processed at Duke–National University of Singapore underwent whole-genome sequencing as described (9). Using DENV-1 isolates from Sri Lanka obtained from dengue cases in 1983, 1984, 1997, 2003, and 2004 (7) and representative DENV-1 sequences for the 4 genotypes, we constructed a phylogenetic tree by using MEGA5 software (10) (Figure; Table).The 4 DENV serotypes found in Sri Lanka have been classifi ed into genotypes according to the nomenclature described by Rico-Hesse (11). The earliest isolates found in 1983 and 1984 belong to South Pacifi c genotype III. More recent isolates obtained during surveillance efforts during 1997–2004 belong to Africa/America genotype IV, indicating that at some point between the early 1980s and the mid 1990s, there was a DENV-1 genotype shift. Analysis of viruses isolated in 2009 indicated that another Asia genotype I of DENV-1 has been introduced into Sri Lanka (Figure) (7). This Asia genotype I virus appears to be responsible for the 2009 epidemic of dengue fever and DHF.ConclusionsA feature of the epidemiology of dengue in Sri Lanka was the lack of DHF in the early 1980s and the increase in the number of severe dengue cases since 1989, more so after 2000. This fi nding was observed despite seroprevalence rates remaining largely the same over time as reported in a previous study (12) and in the current PDVI study (13).Previous epidemics (1989 and 2002–2004) showed a correlation with evolution of DENV-3 genotype III in Sri Lanka, where emergence of new clades of DENV-3 genotype 3 showed a correlation with large increases in the number of reported cases and the geographic range of the virus (7,8). A similar observation was reported for Puerto Rico by Bennett et al., who compared data for DEN2 and DEN4 over 20 years and found that dominant clades were replaced by viral subpopulations existing within the population (14) and in the South Pacifi c region for DENV-2, where a similar clade replacement occurred (15). These clade changes were accompanied by positive selection in the nonstructural protein 2A (NS-2A) gene for DENV-4 and the envelope, premembrane, NS-2A, and NS-4A genes for DENV-2.Our results indicate that introduction of a new DENV-1 genotype coincided with the 2009 dengue epidemic in Sri Lanka. Studies are underway to determine if the proportion of DENV-1 cases in 2009 was greater than in previous years and to assess the role of this new DENV-1 genotype in the severe epidemic of 2009. Further studies are needed to determine if this new genotype has spread to other countries in the region.AcknowledgmentsWe thank the Public Health Department of the Colombo Municipal Council and the Lady Ridgeway Hospital for Children, Colombo, Sri Lanka, for support.This study was supported by the Bill and Melinda Gates Foundation through the PDVI (grant no. 23197).Dr Tissera is a medical epidemiologist at the Epidemiology Unit, Ministry of Health, Colombo, Sri Lanka. His research DISPATCHES2054 Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 17, No. 11, November 2011Table. Dengue virus type 1 strains used in analysis of new dengue virus genotype, Colombo, Sri Lanka Virus strain* Location Subtype Year isolated GenBank accession no. DV1_Aust_1983 Australia III 1983 AB074761 DV1_West_Pacific_1974 Western Pacific III 1974 U88535 DV1_Brazil_1990 Brazil IV 1990 AF226685 DV1_China_1980 People’s Republic of China I 1980 AF350498 DV1_Japan_43 Japan I 1943 AB074760 DV1_Malaysia_72 Malaysia Sylvatic 1972 EF457905 DV1_Seychelles_2004 Seychelles III 2004 DQ285561 DV1_SL_1983a Sri Lanka III 1983 FJ225443 DV1_SL_1984 Sri Lanka III 1984 FJ225444 DV1_SL_1992a Sri Lanka IV 1992 FJ225445 DV1_SL_1997a Sri Lanka IV 1997 FJ225446 DV1_SL_2003a Sri Lanka IV 2003 FJ225447 DV1_SL_2004a Sri Lanka IV 2004 FJ225448 DV1_SL_2004b Sri Lanka IV 2004 FJ225449 DV1_SL_2004c Sri Lanka IV 2004 FJ225450 DV1_Thailand_64 Thailand II 1964 AF180818 DV1_SL_2009a Sri Lanka I 2009 HQ891313 DV1_SL_2009b Sri Lanka I 2009 HQ891314 DV1_SL_2009c Sri Lanka I 2009 HQ891315 DV1_SL_2009e Sri Lanka I 2009 JN054256 DV1_SL_2010b Sri Lanka I 2010 JN054255 *Strains are indicated as genotype_location_year. New Dengue Virus Type 1 Genotype in Sri Lankainterests are the epidemiology, virology, surveillance, and control of dengue.References  1.  Halstead SB. Dengue. Lancet. 2007;370:1644–52. doi:10.1016/S0140-6736(07)61687-0  2.  Gubler DJ. The global emergence/resurgence of arboviral dis-ease as public health problems. Arch Med Res. 2002;33:330–42. doi:10.1016/S0188-4409(02)00378-8  3.  World Health Organization. Dengue. Guidelines for diagnosis, treat-ment, prevention, and control. TDR-WHO [cited 2011 Jul 29]. http://apps.who.int/tdr/svc/publications/training-guideline-publications/dengue-diagnosis-treatment  4.  Kurane I. Dengue hemorrhagic fever with special emphasis on immunopathogenesis. Comp Immunol Microbiol Infect Dis. 2007;30:329–40. doi:10.1016/j.cimid.2007.05.010  5.  Vitarana T, Jayakura WS, Withane N. Historical account of dengue hemorrhagic fever in Sri Lanka. Dengue Bulletin. 1997;21:117–8.  6.  Distribution of suspected DF/DHF by week, Sri Lanka 2004–2010. Epidemiology Unit, Ministry of Health, Sri Lanka [cited 2011 Jul 29]. http://www.epid.gov.lk/pdf/dhf-2010/week31-2010-updated-18-08-2010.pdf  7.  Kanakaratne N, Wahala MP, Messer WB, Tissera HA, Shahani A, Abeysinghe N, et al. Severe dengue epidemics in Sri Lanka, 2003–2006. Emerg Infect Dis. 2009;15:192–9. doi:10.3201/eid1502.080926  8.  Messer WB, Gubler DG, Harris E, Sivananthan K, de Silva AM. Emergence and global spread of a dengue serotype 3, subtype III virus. Emerg Infect Dis. 2003;9:800–9.  9.  Christenbury JG, Aw PP, Ong SH, Schreiber MJ, Chow A, Gubler DJ, et al. A method for full genome sequencing of all four sero-types of the dengue virus. J Virol Methods. 2010;169:202–6. doi:10.1016/j.jviromet.2010.06.01310.  Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: Molecular Evolutionary Genetics Analysis using maxi-mum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol. 2011:28:2731–9. doi:10.1093/molbev/msr12111.  Rico-Hesse R. Microevolution and virulence of dengue viruses. Adv Virus Res. 2003;59:315–41.  doi:10.1016/S0065-3527(03)59009-112.  Messer WB, Vitarana UT, Sivananthan K, Elvtigala J, Preethimala LD, Ramesh R, et al. Epidemiology of dengue in Sri Lanka before and after the emergence of epidemic dengue hemorrhagic fever. Am J Trop Med Hyg. 2002;66:765–73.13.  Tissera HA, De Silva AD, Abeysinghe MR, de Silva AM, Paliha-wadana P, Gunasena S, et al. Third Vaccine Global Congress, 2009. Dengue surveillance in Colombo, Sri Lanka: baseline seropreva-lence among children. Procedia in Vaccinology. 2010;2:109–12 [cit-ed 2011 Sep 29]. http://www.sciencedirect.com/science/article/pii/S1877282X1000021414.  Bennett SN, Holmes EC, Chirivella M, Rodriguez DM, Beltran M, Vorndam V, et al. Selection-driven evolution of emergent dengue virus. Mol Biol Evol. 2003;20:1650–8. doi:10.1093/molbev/msg18215.  Steel A, Gubler DJ, Bennett SN. Natural attenuation of dengue virus type-2 after a series of island outbreaks: a retrospective phylogenetic study of events in the South Pacifi c three decades ago. Virology. 2010;405:505–12. doi:10.1016/j.virol.2010.05.033Address for correspondence: Aruna Dharshan De Silva, Genetech Research Institute, 54 Kitulwatte Rd, Colombo 08, Sri Lanka; email: dslv90@yahoo.com Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 17, No. 11, November 2011 2055

Tissera, Hasitha A

Gubler, Duane J

Wahala, Wahala MPB

de Silva, Aravinda M

Abeysinghe, MR Nihal

Tam, Clarence C

Letson, G William

Margolis, Harold S

LSHTM Research Online

Tissera, HA; Ooi, EE; Gubler, DJ; Tan, Y; Logendra, B; Wahala,W; de Silva, AM; Abeysinghe, MRN; Palihawadana, P; Gunasena, S;Tam, CC; Amarasinghe, A; Letson, GW; Margolis, HS; de Silva, AD(2011) New Dengue Virus Type 1 Genotype in Colombo, Sri Lanka.Emerging infectious diseases, 17 (11). pp. 2053-2055. ISSN 1080-6040 DOI: 10.3201/eid1711.101893Downloaded from: http://researchonline.lshtm.ac.uk/38047/DOI: 10.3201/eid1711.101893Usage GuidelinesPlease refer to usage guidelines at http://researchonline.lshtm.ac.uk/policies.html or alterna-tively contact researchonline@lshtm.ac.uk.Available under license: http://creativecommons.org/licenses/by/2.5/New Dengue Virus Type 1 Genotype in Colombo, Sri LankaHasitha A. Tissera, Eng Eong Ooi, Duane J. Gubler, Ying Tan, Barathy Logendra, Wahala M.P.B. Wahala, Aravinda M. de Silva, M.R. Nihal Abeysinghe, Paba Palihawadana, Sunethra Gunasena, Clarence C. Tam, Ananda Amarasinghe, G. William Letson, Harold S. Margolis, and Aruna Dharshan De SilvaThe number of cases and severity of disease associated with dengue infection in Sri Lanka has been increasing since 1989, when the fi rst epidemic of dengue hemorrhagic fever was recorded. We identifi ed a new dengue virus 1 strain circulating in Sri Lanka that coincided with the 2009 dengue epidemic.Dengue virus (DENV) is a fl avivirus transmitted by Aedes spp. mosquitoes. There are 4 distinct DENV serotypes (DENV-1–4). Infection with a single serotype leads to long-term protective immunity against the homologous serotype but not against other serotypes (1). Globally, dengue is an emerging disease that causes an estimated 50–100 million infections, 500,000 dengue hemorrhagic fever (DHF) cases, and 22,000 deaths annually (2,3).Epidemiologic and other studies indicate that risk factors for severe dengue include secondary infection with a heterologous serotype, the strain of infecting virus, and age and genetic background of the host. Studies are under way to further explore the role of these factors in severe disease (1,4).In Sri Lanka, serologically confi rmed dengue was fi rst reported in 1962 (5), but although all 4 virus serotypes were present and there were cases of DHF, only since 1989 has DHF been considered endemic to Sri Lanka (5). Dengue was made a reportable disease in Sri Lanka in 1996, and the largest epidemic (35,008 reported cases, 170 cases/100,000 population, and 346 deaths) occurred in 2009 (6). DHF epidemics in 1989 and 2002–2004 were associated with emergence of new clades of DENV-3 (7,8). We report a new DENV-1 genotype introduced to Sri Lanka before the 2009 epidemic.The StudyThe study was approved by the Ethical Review Committee of the Faculty of Medicine, University of Colombo, Sri Lanka, and the Institutional Research Board of the International Vaccine Institute, Seoul, South Korea. Serum samples were obtained in 2009 and early 2010 from patients as part of a Pediatric Dengue Vaccine Initiative (PDVI) fever surveillance study in Colombo, Sri Lanka. Samples were originally tested for dengue by reverse transcription PCR at Genetech Research Institute (Colombo, Sri Lanka). A random subset of dengue-positive samples of all 4 serotypes was sent to the Program in Emerging Infectious Diseases Laboratory at Duke–National University of Singapore Graduate Medical School, Singapore, for virus isolation and sequencing.RNA was extracted from virus isolates, subjected to standard reverse transcription PCR to confi rm the presence  Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 17, No. 11, November 2011 2053Author affi liations: Ministry of Health, Colombo, Sri Lanka (H.A. Tissera, M.R.N. Abeysinghe, P. Palihawadana); London School of Hygiene and Tropical Medicine, London, UK (H.A. Tissera, C.C. Tam); Duke–National University of Singapore Graduate Medical School, Singapore (E.E. Ooi, D.J. Gubler, Y. Tan); Genetech Research Institute, Colombo (B. Logendra, A.D. De Silva); University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA (W.M.P.B. Wahala, A.M. de Silva); Medical Research Institute, Colombo (S. Gunasena); and International Vaccine Institute, Seoul, South Korea (A. Amerasinghe, G.W. Letson, H.S. Margolis)DOI: http://dx.doi.org/10.3201/eid1711.101893Figure. Phylogenetic tree of dengue virus 1 (DENV-1) serotype viruses from Sri Lanka (SL), 2009–2010, and other DENV-1 viruses. The tree is based on a 498-bp (nt 2056–2554) fragment that encodes portions of the envelope protein and nonstructural protein 1. Phylogenetic analysis was conducted by using MEGA5 (10). Percentages of replicate trees in which the associated taxa clustered in the bootstrap test (1,000 replicates) are shown next to the branches. Genotype I (Asia) includes SL isolates from 2009–2010; genotype III (South Pacifi c) includes SL isolates from the early 1980s; genotype IV (Africa/Americas) includes SL isolates from the 1990s and early 2000s. Classifi cation and naming of DENV-1 genotypes are based on the report by Rico-Hesse (11). DV, dengue virus. Scale bar indicates nucleotide substitutions per site.of dengue virus, and serotyped as described (7). Samples processed at Duke–National University of Singapore underwent whole-genome sequencing as described (9). Using DENV-1 isolates from Sri Lanka obtained from dengue cases in 1983, 1984, 1997, 2003, and 2004 (7) and representative DENV-1 sequences for the 4 genotypes, we constructed a phylogenetic tree by using MEGA5 software (10) (Figure; Table).The 4 DENV serotypes found in Sri Lanka have been classifi ed into genotypes according to the nomenclature described by Rico-Hesse (11). The earliest isolates found in 1983 and 1984 belong to South Pacifi c genotype III. More recent isolates obtained during surveillance efforts during 1997–2004 belong to Africa/America genotype IV, indicating that at some point between the early 1980s and the mid 1990s, there was a DENV-1 genotype shift. Analysis of viruses isolated in 2009 indicated that another Asia genotype I of DENV-1 has been introduced into Sri Lanka (Figure) (7). This Asia genotype I virus appears to be responsible for the 2009 epidemic of dengue fever and DHF.ConclusionsA feature of the epidemiology of dengue in Sri Lanka was the lack of DHF in the early 1980s and the increase in the number of severe dengue cases since 1989, more so after 2000. This fi nding was observed despite seroprevalence rates remaining largely the same over time as reported in a previous study (12) and in the current PDVI study (13).Previous epidemics (1989 and 2002–2004) showed a correlation with evolution of DENV-3 genotype III in Sri Lanka, where emergence of new clades of DENV-3 genotype 3 showed a correlation with large increases in the number of reported cases and the geographic range of the virus (7,8). A similar observation was reported for Puerto Rico by Bennett et al., who compared data for DEN2 and DEN4 over 20 years and found that dominant clades were replaced by viral subpopulations existing within the population (14) and in the South Pacifi c region for DENV-2, where a similar clade replacement occurred (15). These clade changes were accompanied by positive selection in the nonstructural protein 2A (NS-2A) gene for DENV-4 and the envelope, premembrane, NS-2A, and NS-4A genes for DENV-2.Our results indicate that introduction of a new DENV-1 genotype coincided with the 2009 dengue epidemic in Sri Lanka. Studies are underway to determine if the proportion of DENV-1 cases in 2009 was greater than in previous years and to assess the role of this new DENV-1 genotype in the severe epidemic of 2009. Further studies are needed to determine if this new genotype has spread to other countries in the region.AcknowledgmentsWe thank the Public Health Department of the Colombo Municipal Council and the Lady Ridgeway Hospital for Children, Colombo, Sri Lanka, for support.This study was supported by the Bill and Melinda Gates Foundation through the PDVI (grant no. 23197).Dr Tissera is a medical epidemiologist at the Epidemiology Unit, Ministry of Health, Colombo, Sri Lanka. His research DISPATCHES2054 Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 17, No. 11, November 2011Table. Dengue virus type 1 strains used in analysis of new dengue virus genotype, Colombo, Sri Lanka Virus strain* Location Subtype Year isolated GenBank accession no. DV1_Aust_1983 Australia III 1983 AB074761 DV1_West_Pacific_1974 Western Pacific III 1974 U88535 DV1_Brazil_1990 Brazil IV 1990 AF226685 DV1_China_1980 People’s Republic of China I 1980 AF350498 DV1_Japan_43 Japan I 1943 AB074760 DV1_Malaysia_72 Malaysia Sylvatic 1972 EF457905 DV1_Seychelles_2004 Seychelles III 2004 DQ285561 DV1_SL_1983a Sri Lanka III 1983 FJ225443 DV1_SL_1984 Sri Lanka III 1984 FJ225444 DV1_SL_1992a Sri Lanka IV 1992 FJ225445 DV1_SL_1997a Sri Lanka IV 1997 FJ225446 DV1_SL_2003a Sri Lanka IV 2003 FJ225447 DV1_SL_2004a Sri Lanka IV 2004 FJ225448 DV1_SL_2004b Sri Lanka IV 2004 FJ225449 DV1_SL_2004c Sri Lanka IV 2004 FJ225450 DV1_Thailand_64 Thailand II 1964 AF180818 DV1_SL_2009a Sri Lanka I 2009 HQ891313 DV1_SL_2009b Sri Lanka I 2009 HQ891314 DV1_SL_2009c Sri Lanka I 2009 HQ891315 DV1_SL_2009e Sri Lanka I 2009 JN054256 DV1_SL_2010b Sri Lanka I 2010 JN054255 *Strains are indicated as genotype_location_year. New Dengue Virus Type 1 Genotype in Sri Lankainterests are the epidemiology, virology, surveillance, and control of dengue.References  1.  Halstead SB. Dengue. Lancet. 2007;370:1644–52. doi:10.1016/S0140-6736(07)61687-0  2.  Gubler DJ. The global emergence/resurgence of arboviral dis-ease as public health problems. Arch Med Res. 2002;33:330–42. doi:10.1016/S0188-4409(02)00378-8  3.  World Health Organization. Dengue. Guidelines for diagnosis, treat-ment, prevention, and control. TDR-WHO [cited 2011 Jul 29]. http://apps.who.int/tdr/svc/publications/training-guideline-publications/dengue-diagnosis-treatment  4.  Kurane I. Dengue hemorrhagic fever with special emphasis on immunopathogenesis. Comp Immunol Microbiol Infect Dis. 2007;30:329–40. doi:10.1016/j.cimid.2007.05.010  5.  Vitarana T, Jayakura WS, Withane N. Historical account of dengue hemorrhagic fever in Sri Lanka. Dengue Bulletin. 1997;21:117–8.  6.  Distribution of suspected DF/DHF by week, Sri Lanka 2004–2010. Epidemiology Unit, Ministry of Health, Sri Lanka [cited 2011 Jul 29]. http://www.epid.gov.lk/pdf/dhf-2010/week31-2010-updated-18-08-2010.pdf  7.  Kanakaratne N, Wahala MP, Messer WB, Tissera HA, Shahani A, Abeysinghe N, et al. Severe dengue epidemics in Sri Lanka, 2003–2006. Emerg Infect Dis. 2009;15:192–9. doi:10.3201/eid1502.080926  8.  Messer WB, Gubler DG, Harris E, Sivananthan K, de Silva AM. Emergence and global spread of a dengue serotype 3, subtype III virus. Emerg Infect Dis. 2003;9:800–9.  9.  Christenbury JG, Aw PP, Ong SH, Schreiber MJ, Chow A, Gubler DJ, et al. A method for full genome sequencing of all four sero-types of the dengue virus. J Virol Methods. 2010;169:202–6. doi:10.1016/j.jviromet.2010.06.01310.  Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: Molecular Evolutionary Genetics Analysis using maxi-mum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol. 2011:28:2731–9. doi:10.1093/molbev/msr12111.  Rico-Hesse R. Microevolution and virulence of dengue viruses. Adv Virus Res. 2003;59:315–41.  doi:10.1016/S0065-3527(03)59009-112.  Messer WB, Vitarana UT, Sivananthan K, Elvtigala J, Preethimala LD, Ramesh R, et al. Epidemiology of dengue in Sri Lanka before and after the emergence of epidemic dengue hemorrhagic fever. Am J Trop Med Hyg. 2002;66:765–73.13.  Tissera HA, De Silva AD, Abeysinghe MR, de Silva AM, Paliha-wadana P, Gunasena S, et al. Third Vaccine Global Congress, 2009. Dengue surveillance in Colombo, Sri Lanka: baseline seropreva-lence among children. Procedia in Vaccinology. 2010;2:109–12 [cit-ed 2011 Sep 29]. http://www.sciencedirect.com/science/article/pii/S1877282X1000021414.  Bennett SN, Holmes EC, Chirivella M, Rodriguez DM, Beltran M, Vorndam V, et al. Selection-driven evolution of emergent dengue virus. Mol Biol Evol. 2003;20:1650–8. doi:10.1093/molbev/msg18215.  Steel A, Gubler DJ, Bennett SN. Natural attenuation of dengue virus type-2 after a series of island outbreaks: a retrospective phylogenetic study of events in the South Pacifi c three decades ago. Virology. 2010;405:505–12. doi:10.1016/j.virol.2010.05.033Address for correspondence: Aruna Dharshan De Silva, Genetech Research Institute, 54 Kitulwatte Rd, Colombo 08, Sri Lanka; email: dslv90@yahoo.com Emerging Infectious Diseases • www.cdc.gov/eid • Vol. 17, No. 11, November 2011 2055

New dengue virus type 1 genotype in Colombo, Sri Lanka.

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New Dengue Virus Type 1 Genotype in Colombo, Sri Lanka

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