Molecular Epidemiological Study of Hand, Foot, and Mouth Disease in a Kindergarten-Based Setting in Bangkok, Thailand
Abstract
:1. Introduction
2. Results
2.1. Characteristics of HFMD Cases
2.2. Identification of Enteroviruses in Index Cases and Contact Persons
2.3. Spreading Pattern of HFMD in Kindergartens
2.4. Phylogenetic Analysis of Enteroviruses
3. Discussion
4. Materials and Methods
4.1. Ethics Statement
4.2. Study Site
4.3. Study Design and Specimen Collection
4.4. Sample Preparation and RNA Extraction
4.5. Detection of Enteroviral 5′-UTR Sequences Using qRT-PCR
4.6. Amplification of Enteroviral 5′-UTR Sequences Using RT-PCR, DNA Sequencing, and Phylogenetic Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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School | Number | Sex | Grade | ||||
---|---|---|---|---|---|---|---|
M | F | Nursery | K 1 | K 2 | K 3 | ||
1 | 7 | 4 | 3 | 0 | 5 | 2 | 0 |
2 | 7 | 3 | 4 | 0 | 3 | 4 | 0 |
3 | 4 | 2 | 2 | 0 | 3 | 1 | 0 |
4 | 1 | 0 | 1 | 0 | 1 | 0 | 0 |
5 | 3 | 2 | 1 | 1 | 1 | 1 | 0 |
Total | 22 | 11 | 11 | 1 | 13 | 8 | 0 |
Primer Name | Sequence (5′–3′) |
---|---|
EQ1 (Forward) | 5′-ACATGGTGTGAAGAGTCTATTGAGCT-3′ |
EQ2 (Reverse) | 5′-CCAAAGTAGTCGGTTCCGC-3′ |
EPmod probe (FAM) | 5′-ATTAGCCGCATTCAGGGGCCGGA-3′ |
UniEV_5UTR-F76 (Forward) | 5′-GDAYCTTTGTGCGCCTGTT-3′ |
UniEv_5UTR-F644 (Reverse) | 5′-GCCAATCCAATAGCTATATGG-3′ |
UniEV_5UTR-F172 (Forward) | 5′-GRTCAAGCACTTCTGTHTCC -3′ |
UniEv_5UTR-F610 (Reverse) | 5′-ATTGTCACCATAAGCAGCCA-3′ |
RT-PCR | Nested PCR | ||
---|---|---|---|
Component | Final Concentration | Component | Final Concentration |
2× reaction mix | 1× | 2× PCR buffer (Vivantis) | 1× |
Forward primer | 0.25 µM | Forward primer | 0.25 µM |
Reverse primer | 0.25 µM | Reverse primer | 0.25 µM |
SS III RT/mix platinum Taq | 0.8 µL | Vivantis Taq DNA polymerase | 0.2 µL |
100 mM DTT | 5 mM | nuclease-free water | 6.8 µL |
nuclease-free water | 3.2 µL | 1:50 diluted template | 2 µL |
RNA template | 5 µL |
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Thammasonthijarern, N.; Kosoltanapiwat, N.; Nuprasert, W.; Sittikul, P.; Sriburin, P.; Pan-ngum, W.; Maneekan, P.; Hataiyusuk, S.; Hattasingh, W.; Thaipadungpanit, J.; et al. Molecular Epidemiological Study of Hand, Foot, and Mouth Disease in a Kindergarten-Based Setting in Bangkok, Thailand. Pathogens 2021, 10, 576. https://doi.org/10.3390/pathogens10050576
Thammasonthijarern N, Kosoltanapiwat N, Nuprasert W, Sittikul P, Sriburin P, Pan-ngum W, Maneekan P, Hataiyusuk S, Hattasingh W, Thaipadungpanit J, et al. Molecular Epidemiological Study of Hand, Foot, and Mouth Disease in a Kindergarten-Based Setting in Bangkok, Thailand. Pathogens. 2021; 10(5):576. https://doi.org/10.3390/pathogens10050576
Chicago/Turabian StyleThammasonthijarern, Nipa, Nathamon Kosoltanapiwat, Warisa Nuprasert, Pichamon Sittikul, Pimolpachr Sriburin, Wirichada Pan-ngum, Pannamas Maneekan, Somboon Hataiyusuk, Weerawan Hattasingh, Janjira Thaipadungpanit, and et al. 2021. "Molecular Epidemiological Study of Hand, Foot, and Mouth Disease in a Kindergarten-Based Setting in Bangkok, Thailand" Pathogens 10, no. 5: 576. https://doi.org/10.3390/pathogens10050576