Molecular Epidemiology and Evolution of Human Enterovirus Serotype 68 in Thailand, 2006–2011

BACKGROUND: Publications worldwide have reported on the re-occurrence of human enterovirus 68 (EV68), a rarely detected pathogen usually causing respiratory illness. However, epidemiological data regarding this virus in particular on the Asian continent has so far been limited. METHODOLOGY/FINDINGS: We investigated the epidemiology and genetic variability of EV68 infection among Thai children with respiratory illnesses from 2006-2011 (n = 1810). Semi-nested PCR using primer sets for amplification of the 5'-untranslated region through VP2 was performed for rhino-enterovirus detection. Altogether, 25 cases were confirmed as EV68 infection indicating a prevalence of 1.4% in the entire study population. Interestingly, the majority of samples were children aged >5 years (64%). Also, co-infection with other viruses was found in 28%, while pandemic H1N1 influenza/2009 virus was the most common co-infection. Of EV68-positive patients, 36% required hospitalizations with the common clinical presentations of fever, cough, dyspnea, and wheezing. The present study has shown that EV68 was extremely rare until 2009 (0.9%). An increasing annual prevalence was found in 2010 (1.6%) with the highest detection frequency in 2011 (4.3%). Based on analysis of the VP1 gene, the evolutionary rate of EV68 was estimated at 4.93 × 10(-3) substitutions/site/year. Major bifurcation of the currently circulating EV68 strains occurred 66 years ago (1945.31 with (1925.95-1960.46)95% HPD). Among the current lineages, 3 clusters of EV68 were categorized based on the different molecular signatures in the BC and DE loops of VP1 combined with high posterior probability values. Each cluster has branched off from their common ancestor at least 36 years ago (1975.78 with (1946.13-1984.97)95% HPD). CONCLUSION: Differences in epidemiological characteristic and seasonal profile of EV68 have been found in this study. Results from Bayesian phylogenetic investigations also revealed that EV68 should be recognized as a genetically diverse virus with a substitution rate identical to that of enterovirus 71 genotype B (4.2 × 10(-3 )s/s/y)

Infantile osteoarticular tuberculosis misdiagnosed as Bacillus Calmette-Guerin related osteomyelitis

Tuberculosis, a re-emerging public health problem, is uncommon in infancy. Two healthy completely immunized infants presenting with manifestations compatible with osteoarticular infection required surgical debridement. The cultures of the specimens were positive for M. tuberculosis (MTB) complex comprised multiple subspecies. One case was misdiagnosed as a Bacillus Calmette-Guerin (BCG) related osteomyelitis by a polymerase chain reaction (PCR) based on detection of genes at the region of difference 1. Genome extraction and PCR using the rimM gene and sequences analysis against MTB and BCG control samples confirmed that both specimens were infected by M. tuberculosis. The lesions were successfully healed within one year. Surgical debridement of suspected lesions is warranted in infants as a definitive treatment and to obtain tissues for further evaluation. Microbiological cultures only confirm nonspecific MTB complex infection. PCR kits may yield a false positive result. Identification of the pathogen by DNA extraction and sequence analysis should be recommended

Infantile osteoarticular tuberculosis misdiagnosed as Bacillus Calmette-Guerin related osteomyelitis

Tuberculosis, a re-emerging public health problem, is uncommon in infancy. Two healthy completely immunized infants presenting with manifestations compatible with osteoarticular infection required surgical debridement. The cultures of the specimens were positive for M. tuberculosis (MTB) complex comprised multiple subspecies. One case was misdiagnosed as a Bacillus Calmette-Guerin (BCG) related osteomyelitis by a polymerase chain reaction (PCR) based on detection of genes at the region of difference 1. Genome extraction and PCR using the rimM gene and sequences analysis against MTB and BCG control samples confirmed that both specimens were infected by M. tuberculosis. The lesions were successfully healed within one year. Surgical debridement of suspected lesions is warranted in infants as a definitive treatment and to obtain tissues for further evaluation. Microbiological cultures only confirm nonspecific MTB complex infection. PCR kits may yield a false positive result. Identification of the pathogen by DNA extraction and sequence analysis should be recommended

Enterovirus A71 Infection, Thailand, 2017

An outbreak of hand, foot and mouth disease among children in Thailand peaked in August 2017. Enterovirus A71 subgenogroup B5 caused most (33.8%, 163/482) cases. Severe disease (myocarditis and encephalitis) was observed in 1 patient. Coxsackievirus A6 was detected in 6.0% (29/482) of patients, and coxsackievirus A16 was detected in 2.7% (13/482) of patients

Climate factors influence seasonal influenza activity in Bangkok, Thailand.

Yearly increase in influenza activity is associated with cold and dry winter in the temperate regions, while influenza patterns in tropical countries vary significantly by regional climates and geographic locations. To examine the association between influenza activity in Thailand and local climate factors including temperature, relative humidity, and rainfall, we analyzed the influenza surveillance data from January 2010 to December 2018 obtained from a large private hospital in Bangkok. We found that approximately one in five influenza-like illness samples (21.6% or 6,678/30,852) tested positive for influenza virus. Influenza virus typing showed that 34.2% were influenza A(H1N1)pdm09, 46.0% were influenza A(H3N2), and 19.8% were influenza B virus. There were two seasonal waves of increased influenza activity. Peak influenza A(H1N1)pdm09 activity occurred in February and again in August, while influenza A(H3N2) and influenza B viruses were primarily detected in August and September. Time series analysis suggests that increased relative humidity was significantly associated with increased influenza activity in Bangkok. Months with peak influenza activity generally followed the most humid months of the year. We performed the seasonal autoregressive integrated moving average (SARIMA) multivariate analysis of all influenza activity on the 2011 to 2017 data to predict the influenza activity for 2018. The resulting model closely resembled the actual observed overall influenza detected that year. Consequently, the ability to predict seasonal pattern of influenza in a large tropical city such as Bangkok may enable better public health planning and underscores the importance of annual influenza vaccination prior to the rainy season

Respiratory syncytial virus genotypes NA1, ON1, and BA9 are prevalent in Thailand, 2012–2015

Respiratory syncytial virus (RSV) causes acute lower respiratory tract infection in infants and young children worldwide. To investigate the RSV burden in Thailand over four consecutive years (January 2012 to December 2015), we screened 3,306 samples obtained from children ≤5 years old with acute respiratory tract infection using semi-nested reverse-transcription polymerase chain reaction (RT-PCR). In all, 8.4% (277/3,306) of the specimens tested positive for RSV, most of which appeared in the rainy months of July to November. We then genotyped RSV by sequencing the G glycoprotein gene and performed phylogenetic analysis to determine the RSV antigenic subgroup. The majority (57.4%, 159/277) of the RSV belonged to subgroup A (RSV-A), of which NA1 genotype was the most common in 2012 while ON1 genotype became prevalent the following year. Among samples tested positive for RSV-B subgroup B (RSV-B) (42.6%, 118/277), most were genotype BA9 (92.6%, 87/94) with some BA10 and BA-C. Predicted amino acid sequence from the partial G region showed highly conserved N-linked glycosylation site at residue N237 among all RSV-A ON1 strains (68/68), and at residues N296 (86/87) and N310 (87/87) among RSV-B BA9 strains. Positive selection of key residues combined with notable sequence variations on the G gene contributed to the continued circulation of this rapidly evolving virus

Evolutionary and Genetic Recombination Analyses of Coxsackievirus A6 Variants Associated with Hand, Foot, and Mouth Disease Outbreaks in Thailand between 2019 and 2022

Coxsackievirus (CV)-A6 infections cause hand, foot, and mouth disease (HFMD) in children and adults. Despite the serious public health threat presented by CV-A6 infections, our understanding of the mechanisms by which new CV-A6 strains emerge remains limited. This study investigated the molecular epidemiological trends, evolutionary dynamics, and recombination characteristics of CV-A6-associated HFMD in Thailand between 2019 and 2022. In the HFMD patient samples collected during the 4-year study period, we identified enterovirus (EV) RNA in 368 samples (48.7%), of which CV-A6 (23.7%) was the predominant genotype, followed by CV-A4 (6%), EV-A71 (3.7%), and CV-A16 (3.4%). According to the partial viral protein (VP) 1 sequences, all these CV-A6 strains belonged to the D3 clade. Based on the viral-RNA-dependent RNA polymerase (RdRp) gene, four recombinant forms (RFs), RF-A (147, 84.5%), RF-N (11, 6.3%), RF-H (1, 0.6%), and newly RF-Y (15, 8.6%), were identified throughout the study period. Results from the similarity plot and bootscan analyses revealed that the 3D polymerase (3Dpol) region of the D3/RF-Y subclade consists of sequences highly similar to CV-A10. We envisage that the epidemiological and evolutionarily insights presented in this manuscript will contribute to the development of vaccines to prevent the spread of CV-A6 infection

Hand, Foot and Mouth Disease Caused by Coxsackievirus A6, Thailand, 2012

In Thailand, hand, foot, and mouth disease (HFMD) is usually caused by enterovirus 71 or coxsackievirus A16. To determine the cause of a large outbreak of HFMD in Thailand during June–August 2012, we examined patient specimens. Coxsackievirus A6 was the causative agent. To improve prevention and control, causes of HFMD should be monitored