Multiple chikungunya virus introductions in Lao PDR from 2014 to 2020.

The first documented chikungunya virus (CHIKV) outbreak in Lao People's Democratic Republic (Lao PDR) occurred in 2012-2013. Since then, several imported and a few autochthonous cases were identified by the national arbovirus surveillance network. The present study aimed to summarize the main genetic features of the CHIKV strains detected in Lao PDR between 2014 and 2020. Samples from Lao patients presenting symptoms compatible with a CHIKV infection were centralized in Vientiane Capital city for real-time RT-PCR screening. Molecular epidemiology was performed by sequencing the E2-6K-E1 region. From 2014 to 2020, two Asian lineage isolates (e.g. French Polynesia; Indonesia), one ECSA-IOL lineage isolate (e.g. Thailand) and one unclassified (e.g. Myanmar) were imported in Vientiane Capital city. Sequences from the autochthonous cases recorded in the Central and Southern parts of the country between July and September 2020 belonged to the ECSA-IOL lineage and clustered with CHIKV strains recently detected in neighboring countries. These results demonstrate the multiple CHIKV introductions in Lao PDR since 2014 and provide evidence for sporadic and time-limited circulation of CHIKV in the country. Even if the circulation of CHIKV seems to be geographically and temporally limited in Lao PDR, the development of international tourism and trade may cause future outbreaks of CHIKV in the country and at the regional level.

Low Transmission of Chikungunya Virus by Aedes aegypti from Vientiane Capital, Lao PDR

In 2012−2013, chikungunya virus (CHIKV) was the cause of a major outbreak in the southern part of Lao People's Democratic Republic (Lao PDR). Since then, only a few imported cases, with isolates belonging to different lineages, were recorded between 2014 and 2020 in Vientiane capital and few autochthonous cases of ECSA-IOL lineage were detected in the south of the country in 2020. The CHIKV epidemiological profile contrasts with the continuous and intensive circulation of dengue virus in the country, especially in Vientiane capital. The study's aim was to investigate the ability of the local field-derived Aedes aegypti population from Vientiane capital to transmit the Asian and ECSA-IOL lineages of CHIKV. Our results revealed that, for both CHIKV lineages, infection rates were low and dissemination rates were high. The transmission rates and efficiencies evidenced a low vector competence for the CHIKV tested. Although this population of Ae. aegypti showed a relatively modest vector competence for these two CHIKV lineages, several other factors could influence arbovirus emergence such as the longevity and density of female mosquitoes. Due to the active circulation of CHIKV in Southeast Asia, investigations on these factors should be done to prevent the risk of CHIKV emergence and spread in Lao PDR and neighboring countries.

Using Background Sequencing Data to Anticipate DENV-1 Circulation in the Lao PDR.

Since its first detection in 1979, dengue fever has been considered a major public health issue in the Lao People's Democratic Republic (PDR). Dengue virus (DENV) serotype 1 was the cause of an epidemic in 2010-2011. Between 2012 and 2020, major outbreaks due successively to DENV-3, DENV-4 and recently DENV-2 have been recorded. However, DENV-1 still co-circulated in the country over this period. Here, we summarize epidemiological and molecular data of DENV-1 between 2016 and 2020 in the Lao PDR. Our data highlight the continuous circulation of DENV-1 in the country at levels ranging from 16% to 22% among serotyping tests. In addition, the phylogenetic analysis has revealed the circulation of DENV-1 genotype I at least since 2008 with a co-circulation of different clusters. Sequence data support independent DENV-1 introductions in the Lao PDR correlated with an active circulation of this serotype at the regional level in Southeast Asia. The maintenance of DENV-1 circulation over the last ten years supports a low level of immunity against this serotype within the Lao population. Thereby, the risk of a DENV-1 epidemic cannot be ruled out in the future, and this emphasizes the importance of maintaining an integrated surveillance approach to prevent major outbreaks.

First probable case of congenital Zika syndrome in Lao People's Democratic Republic.

Zika virus (ZIKV) is a Flavivirus transmitted by Aedes mosquitoes, and was responsible for a worldwide outbreak between 2013 and 2016. However, no ZIKV outbreak has been described in Southeast Asia since 2017. In this study, we report the first microcephaly case with probable ZIKV infection during pregnancy in Lao People's Democratic Republic.

Trends of the Dengue Serotype-4 Circulation with Epidemiological, Phylogenetic, and Entomological Insights in Lao PDR between 2015 and 2019.

Dengue outbreaks have regularly been recorded in Lao People's Democratic Republic (PDR) since the first detection of the disease in 1979. In 2012, an integrated arbovirus surveillance network was set up in Lao PDR and an entomological surveillance has been implemented since 2016 in Vientiane Capital. Here, we report a study combining epidemiological, phylogenetic, and entomological analyzes during the largest DENV-4 epidemic ever recorded in Lao PDR (2015-2019). Strikingly, from 2015 to 2019, we reported the DENV-4 emergence and spread at the country level after two large epidemics predominated by DENV-3 and DENV-1, respectively, in 2012-2013 and 2015. Our data revealed a significant difference in the median age of the patient infected by DENV-4 compared to the other serotypes. Phylogenetic analysis demonstrated the circulation of DENV-4 Genotype I at the country level since at least 2013. The entomological surveillance showed a predominance of Aedesaegypti compared to Aedesalbopictus and high abundance of these vectors in dry and rainy seasons between 2016 and 2019, in Vientiane Capital. Overall, these results emphasized the importance of an integrated approach to evaluate factors, which could impact the circulation and the epidemiological profile of dengue viruses, especially in endemic countries like Lao PDR.

Rapid genotyping protocol to improve dengue virus serotype 2 survey in Lao PDR.

Dengue fever is one of the major public health problems in Lao PDR. Over the last decade, dengue virus (DENV) epidemics were characterized by a novel predominant serotype accompanied by at least two other serotypes. Since 2008, DENV-2 circulated at a low level in Lao PDR but its epidemiologic profile changed at the end of 2018. Indeed, the number of confirmed DENV-2 cases suddenly increased in October 2018 and DENV-2 became predominant at the country level in early 2019. We developed a Genotype Screening Protocol (GSP) to determine the origin(s) of the Lao DENV-2 and study their genetic polymorphism. With a good correlation with full envelope gene sequencing data, this molecular epidemiology tool evidence the co-circulation of two highly polymorphic DENV-2 genotypes, i.e. Asian I and Cosmopolitan genotypes, over the last five years, suggesting multiple introductions of DENV-2 in the country. GSP approach provides relevant first line information that may help countries with limited laboratory resources to reinforce their capabilities to DENV-2 and to follow the epidemics progresses and assess situations at the regional level.