Limits...
Detection of Zika Virus Infection in Thailand, 2012-2014.

Buathong R, Hermann L, Thaisomboonsuk B, Rutvisuttinunt W, Klungthong C, Chinnawirotpisan P, Manasatienkij W, Nisalak A, Fernandez S, Yoon IK, Akrasewi P, Plipat T - Am. J. Trop. Med. Hyg. (2015)

Bottom Line: Zika virus (ZIKV) is an emerging mosquito-borne pathogen with reported cases in Africa, Asia, and large outbreaks in the Pacific.However, there have been several cases reported in travelers returning from Thailand.These endemic cases, combined with previous reports in travelers, provide evidence that ZIKV is widespread throughout Thailand.

View Article: PubMed Central - PubMed

Affiliation: Department of Disease Control, Bureau of Epidemiology, Ministry of Public Health, Nonthaburi, Thailand; Department of Virology, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand; Department of Medicine, University of Toronto, Toronto, Ontario, Canada.

Show MeSH

Related in: MedlinePlus

Maximum likelihood phylogenetic trees of fragments of Zika virus nonstructural protein 5 (ZIKV NS5) utilizing GTR+G+I model with 14 reference ZIKV strains from GenBank. The contig sequences, obtained from de novo assembly and BLASTn, of KM851039/Thailand/2014 were analyzed against eight references from the African lineage (Accession numbers: KF268948, KF268950, KF268949, LC002520, AY632535, NC012532, HQ234500, and HQ234501) and six references from the Asian lineage (Accession numbers: KJ776791, JN860885, EU545988, HQ234499, KF993678, and KM851038). The scale of genetic distance equal to 0.02 is indicated in the bottom left of the panel. Bootstrap ≥ 70 is demonstrated next to the node of the tree. The year of collection is unknown for a number of African strains. Drawing is not to scale.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4530765&req=5

Figure 1: Maximum likelihood phylogenetic trees of fragments of Zika virus nonstructural protein 5 (ZIKV NS5) utilizing GTR+G+I model with 14 reference ZIKV strains from GenBank. The contig sequences, obtained from de novo assembly and BLASTn, of KM851039/Thailand/2014 were analyzed against eight references from the African lineage (Accession numbers: KF268948, KF268950, KF268949, LC002520, AY632535, NC012532, HQ234500, and HQ234501) and six references from the Asian lineage (Accession numbers: KJ776791, JN860885, EU545988, HQ234499, KF993678, and KM851038). The scale of genetic distance equal to 0.02 is indicated in the bottom left of the panel. Bootstrap ≥ 70 is demonstrated next to the node of the tree. The year of collection is unknown for a number of African strains. Drawing is not to scale.

Mentions: Three samples taken from Ratchaburi province, Thailand, were positive for ZIKV infection based on serologic studies whereas ZIKV RNA was detected in the other four samples. The 77 base pair RT-PCR amplicon, used for ZIKV diagnosis from a sample from Sisaket province, was sequenced and found to have high nucleotide identity (between 90% and 100%) to the prM/E region of ZIKV sequences reported in GenBank. Subsequently, three serum samples collected in Phetchabun province that were positive or equivocal for ZIKV by PCR were sent for virus isolation by intrathoracic inoculation of Toxorhynchites splendens mosquitoes followed by inoculation of mosquito-derived C6/36 cells. Typing ELISA using the universal flavivirus (4G2 monoclonal) antibody and DENV serotypes 1, 2, 3, and 4 specific monoclonal antibodies was also completed on the supernatants from the C6/36 cells.18,19 One supernatant sample positive for 4G2 antibody, but negative for DENV antibodies, underwent next-generation sequencing (Illumina MiSeq Platform, Illumina, Hayward, CA). De novo assembly produced a 10,685 nucleotide (nt) consensus sequence (depth of coverage greater than 10,000). Maximum likelihood (ML) phylogenetic analysis of a 789-bp partial sequence of ZIKV nonstructural protein 5 (ZIKV NS5) (GenBank accession no. KM851039) indicated that the isolate belonged to the ZIKV Asian lineage, which includes strains from the Canadian traveler to Thailand (2013)11 and strains from Cambodia (2010),15 Micronesia (2007),2 and French Polynesia (2013)20 (Figure 1). Pairwise genetic distance calculation illustrated that the isolate was most closely related to the French Polynesia 2013 strain (p-distance = 0.009) with approximately 99% nucleotide identity (782/789).


Detection of Zika Virus Infection in Thailand, 2012-2014.

Buathong R, Hermann L, Thaisomboonsuk B, Rutvisuttinunt W, Klungthong C, Chinnawirotpisan P, Manasatienkij W, Nisalak A, Fernandez S, Yoon IK, Akrasewi P, Plipat T - Am. J. Trop. Med. Hyg. (2015)

Maximum likelihood phylogenetic trees of fragments of Zika virus nonstructural protein 5 (ZIKV NS5) utilizing GTR+G+I model with 14 reference ZIKV strains from GenBank. The contig sequences, obtained from de novo assembly and BLASTn, of KM851039/Thailand/2014 were analyzed against eight references from the African lineage (Accession numbers: KF268948, KF268950, KF268949, LC002520, AY632535, NC012532, HQ234500, and HQ234501) and six references from the Asian lineage (Accession numbers: KJ776791, JN860885, EU545988, HQ234499, KF993678, and KM851038). The scale of genetic distance equal to 0.02 is indicated in the bottom left of the panel. Bootstrap ≥ 70 is demonstrated next to the node of the tree. The year of collection is unknown for a number of African strains. Drawing is not to scale.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4530765&req=5

Figure 1: Maximum likelihood phylogenetic trees of fragments of Zika virus nonstructural protein 5 (ZIKV NS5) utilizing GTR+G+I model with 14 reference ZIKV strains from GenBank. The contig sequences, obtained from de novo assembly and BLASTn, of KM851039/Thailand/2014 were analyzed against eight references from the African lineage (Accession numbers: KF268948, KF268950, KF268949, LC002520, AY632535, NC012532, HQ234500, and HQ234501) and six references from the Asian lineage (Accession numbers: KJ776791, JN860885, EU545988, HQ234499, KF993678, and KM851038). The scale of genetic distance equal to 0.02 is indicated in the bottom left of the panel. Bootstrap ≥ 70 is demonstrated next to the node of the tree. The year of collection is unknown for a number of African strains. Drawing is not to scale.
Mentions: Three samples taken from Ratchaburi province, Thailand, were positive for ZIKV infection based on serologic studies whereas ZIKV RNA was detected in the other four samples. The 77 base pair RT-PCR amplicon, used for ZIKV diagnosis from a sample from Sisaket province, was sequenced and found to have high nucleotide identity (between 90% and 100%) to the prM/E region of ZIKV sequences reported in GenBank. Subsequently, three serum samples collected in Phetchabun province that were positive or equivocal for ZIKV by PCR were sent for virus isolation by intrathoracic inoculation of Toxorhynchites splendens mosquitoes followed by inoculation of mosquito-derived C6/36 cells. Typing ELISA using the universal flavivirus (4G2 monoclonal) antibody and DENV serotypes 1, 2, 3, and 4 specific monoclonal antibodies was also completed on the supernatants from the C6/36 cells.18,19 One supernatant sample positive for 4G2 antibody, but negative for DENV antibodies, underwent next-generation sequencing (Illumina MiSeq Platform, Illumina, Hayward, CA). De novo assembly produced a 10,685 nucleotide (nt) consensus sequence (depth of coverage greater than 10,000). Maximum likelihood (ML) phylogenetic analysis of a 789-bp partial sequence of ZIKV nonstructural protein 5 (ZIKV NS5) (GenBank accession no. KM851039) indicated that the isolate belonged to the ZIKV Asian lineage, which includes strains from the Canadian traveler to Thailand (2013)11 and strains from Cambodia (2010),15 Micronesia (2007),2 and French Polynesia (2013)20 (Figure 1). Pairwise genetic distance calculation illustrated that the isolate was most closely related to the French Polynesia 2013 strain (p-distance = 0.009) with approximately 99% nucleotide identity (782/789).

Bottom Line: Zika virus (ZIKV) is an emerging mosquito-borne pathogen with reported cases in Africa, Asia, and large outbreaks in the Pacific.However, there have been several cases reported in travelers returning from Thailand.These endemic cases, combined with previous reports in travelers, provide evidence that ZIKV is widespread throughout Thailand.

View Article: PubMed Central - PubMed

Affiliation: Department of Disease Control, Bureau of Epidemiology, Ministry of Public Health, Nonthaburi, Thailand; Department of Virology, Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand; Department of Medicine, University of Toronto, Toronto, Ontario, Canada.

Show MeSH
Related in: MedlinePlus