Serologic testing of randomly selected children after hepatitis B vaccination: a cross-sectional population-based study in Lao People's Democratic Republic.

BACKGROUND: Population immunity against hepatitis B virus (HBV) in Lao People's Demographic Republic (PDR) has not been examined since the national HBV vaccination program was started in 2002. Vaccine has been observed to be frozen at times during cold-chain transport in vaccination programs in Lao PDR and other developing countries, which will inactivate the vaccine. Therefore, this study used post-vaccination serologic testing to evaluate the effects of HBV immunization in Lao PDR. METHODS: A cross-sectional serologic study was conducted among children (age range, 5-9 years) and mothers (15-45 years) who were randomly selected using probability-proportional-to-size sampling from central Lao PDR. Blood samples were collected as dried blood spots (DBS) and analyzed using chemiluminescent microparticle immunoassay to detect anti-hepatitis B surface (HBs) titers. We also evaluated the correlation between anti-HBs levels measured in DBS and serum among healthy healthcare workers in Vientiane. RESULTS: Anti-HBs titers from DBS were strongly correlated with serum levels (correlation coefficient = 0.999) in all 12 healthcare workers evaluated. A linear regression model showed that 10 mIU/mL of serum anti-HBs was equivalent to 3.45 mIU/mL (95% CI: 3.06-3.85) of DBS. Among 911 mother-child pairs tested, 171 children had documentation of vaccination. Of the 147 children who had received ≥3 doses of the hepatitis B vaccine, 1 (0.7%) was positive for anti-HBs. The remaining 24 children received the hepatitis B vaccine only twice, once or no dose. CONCLUSIONS: The results showed extremely low positivity for anti-HBs among vaccinated children in central Lao PDR. Therefore, post-vaccination serologic testing is important to evaluate population immunity against HBV infection. DBS testing is a potential low-cost tool to evaluating the effectiveness of HBV vaccination programs.

Analysing the characteristics of a measles outbreak in Houaphanh province to guide measles elimination in the Lao People's Democratic Republic.

INTRODUCTION: In recent years, the incidence of measles has declined in the Lao People's Democratic Republic. However, an outbreak was reported in August 2014 in Houaphanh province, which was the biggest outbreak in the country since 2008. We describe the characteristics of this outbreak and outline critical interventions for the Lao People's Democratic Republic to achieve measles elimination. METHODS: Fever and rash cases in the Khouan and Samtai districts with an onset date from 1 September to 25 October 2014 were investigated. Active case finding and health facility record reviews were carried out. Appropriate samples from the individuals with suspected measles were tested to confirm the diagnosis. RESULTS: A total of 265 suspected cases including 12 deaths were reported from eight villages in the Khouan and Samtai districts. Forty-five individuals tested positive for measles IgM. Most of the confirmed patients were male (n = 28, 62%), less than 5 years old (n = 23, 51%) and from the Hmong ethnic community (n = 44, 98%). The majority of the people with suspected measles (n = 213, 80%) and all the confirmed ones were unvaccinated. A measles vaccination campaign conducted in the eight affected villages resulted in 76% coverage of the targeted population. DISCUSSION: Low routine coverage and measles occurrence among unvaccinated individuals indicate underimmunized areas. The geographical and sociodemographic characteristics of this outbreak highlight the need for tailored vaccination strategies to close the immunity gap. A sensitive surveillance system that is able to detect, notify, investigate and guide response measures, including a second measles dose in the routine immunization schedule, will be essential for the Lao People's Democratic Republic to attain its measles elimination status.

Evaluation of nationwide supplementary immunization in Lao People's Democratic Republic: Population-based seroprevalence survey of anti-measles and anti-rubella IgG in children and adults, mathematical modelling and a stability testing of the vaccine.

BACKGROUND: Measles outbreaks have occurred in some countries despite supplementary immunization activities (SIA) using measles-containing vaccine with high vaccination coverage. We conducted a cross-sectional seroprevalence survey to estimate population immunity in Lao People's Democratic Republic where repeated mass immunization has failed to eliminate measles. METHODS AND FINDINGS: In this nationwide multistage cluster sampling survey conducted in 2014 based on probability proportionate to size sampling, blood samples were collected from 2,135 children and adults living in 52 randomly selected villages. Anti-measles and anti-rubella IgG were measured, and IgG prevalence was calculated. We applied mathematical modelling to estimate the number of cases of congenital rubella syndrome (CRS) in 2013 that were averted by the 2011 SIA. A stability testing was applied to the MR vaccine at 4°C, 25°C, and 35°C to examine stability differences between measles and rubella vaccine components. Measles IgG prevalence was significantly lower in the target age groups (5-21 years) of the 2011 SIA using a combination vaccine for measles and rubella vaccine (MR vaccine) than in young adults (22-39 years) (86.8% [95% CI: 83.0-90.6] vs. 99.0% [98.3-99.8]; p<0.001), whereas rubella IgG prevalence was significantly higher (88.2% [84.5-91.8] vs. 74.6% [70.7-78.5]; p<0.001). In the SIA target age groups, prevalence of measles IgG, but not rubella IgG, increased with age. CRS cases prevented in 2013 ranged from 16 [0-50] to 92 [32-180] if the force of infection had remained unchanged or had been reduced by 75%, respectively. In freeze-dried conditions, the measles vaccine component was more heat sensitive than the rubella component. CONCLUSIONS: Inconsistent IgG prevalence between measles and rubella in Lao PDR can be partly explained by different stability of the measles and rubella vaccine components under heat exposure. Suboptimal vaccine handling may cause insufficient immunogenicity for measles, which subsequently leads to an outbreak despite high SIA coverage, while direct evidence is lacking. Temperature monitoring of the vaccine should be conducted.

Chikungunya virus emergence in the Lao PDR, 2012-2013.

In May 2012, the first authenticated cases of active chikungunya virus infection were detected in Champasak Province, Southern Laos. Analysis of series of human samples and mosquito specimens collected during the outbreak and over the year that followed the emergence enabled the drawing up of a map of the progression of CHIKV and the establishment of a full genetic characterization of the virus.

Antiviral Drug-Resistant Influenza B Viruses Carrying H134N Substitution in Neuraminidase, Laos, February 2016.

In February 2016, three influenza B/Victoria/2/87 lineage viruses exhibiting 4- to 158-fold reduced inhibition by neuraminidase inhibitors were detected in Laos. These viruses had an H134N substitution in the neuraminidase and replicated efficiently in vitro and in ferrets. Current antiviral drugs may be ineffective in controlling infections caused by viruses harboring this mutation.

Trends in the incidence of acute watery diarrhoea in the Lao People's Democratic Republic, 2009-2013.

Diarrhoeal disease is the second leading cause of death in children under age 5 worldwide, with rotavirus being the main etiology. In the Lao People's Democratic Republic, acute watery diarrhoea (AWD) was introduced as one of the national notifiable diseases in 2004. We retrospectively reviewed the aggregate (n = 117 277) and case-based (n = 67 755) AWD surveillance data from 2009 to 2013 reported weekly from 1115 health facilities nationwide. Rotavirus rapid test data from all eight sentinel sites in Vientiane Capital in 2013 were also collected for analysis. The incidence of AWD ranged between 215 and 476 cases per 100 000 population and increased from 2009 to 2012 when it levelled off. The most affected age group was children under 5 who were about seven to nine times more likely to have AWD than the rest of the population (P < 0.0001). In children under 5, 74.8% of the cases were aged 0-24 months and AWD was 1.28 times more common in males (P < 0.0001). Among the 230 stool specimens tested in children under 5 in 2013, 109 (47.4%) tested positive for rotavirus. The increased AWD incidence over the study period may reflect a true increase in AWD or an improved sensitivity of the system. We recommend new mothers breastfeed up to two years after birth, which is known to reduce AWD morbidity and mortality in young children. We also recommend conducting rotavirus disease burden and cost-effectiveness studies to explore the benefits of introduction of rotavirus vaccine.

Region-wide synchrony and traveling waves of dengue across eight countries in Southeast Asia.

Dengue is a mosquito-transmitted virus infection that causes epidemics of febrile illness and hemorrhagic fever across the tropics and subtropics worldwide. Annual epidemics are commonly observed, but there is substantial spatiotemporal heterogeneity in intensity. A better understanding of this heterogeneity in dengue transmission could lead to improved epidemic prediction and disease control. Time series decomposition methods enable the isolation and study of temporal epidemic dynamics with a specific periodicity (e.g., annual cycles related to climatic drivers and multiannual cycles caused by dynamics in population immunity). We collected and analyzed up to 18 y of monthly dengue surveillance reports on a total of 3.5 million reported dengue cases from 273 provinces in eight countries in Southeast Asia, covering ∼ 10(7) km(2). We detected strong patterns of synchronous dengue transmission across the entire region, most markedly during a period of high incidence in 1997-1998, which was followed by a period of extremely low incidence in 2001-2002. This synchrony in dengue incidence coincided with elevated temperatures throughout the region in 1997-1998 and the strongest El Niño episode of the century. Multiannual dengue cycles (2-5 y) were highly coherent with the Oceanic Niño Index, and synchrony of these cycles increased with temperature. We also detected localized traveling waves of multiannual dengue epidemic cycles in Thailand, Laos, and the Philippines that were dependent on temperature. This study reveals forcing mechanisms that drive synchronization of dengue epidemics on a continental scale across Southeast Asia.

Seroprevalence of chronic hepatitis B, as determined from dried blood spots, among children and their mothers in central Lao People's Democratic Republic: a multistage, stratified cluster sampling survey.

BACKGROUND: There is limited information regarding the prevalence of hepatitis B in Lao PDR, where the hepatitis disease burden is substantial. Thus, reliable seroprevalence data is needed for the disease, based on probability sampling. METHODS: A stratified, multistage, cluster sampling survey of hepatitis B surface antigen (HBsAg) positivity among children aged 5-9 years and their mothers aged 15-45 years was conducted. Participants were selected randomly from the central region of Lao PDR via probability-proportional-to-size sampling. Blood samples were collected onto filter paper and subsequently analyzed using a chemiluminescent microparticle immunoassay. RESULTS: A total of 911 mother-and-child pairs were collected; the seroprevalence of HBsAg was estimated to be 2.1% (95% confidence interval 0.8-3.4%) among children and 4.1% (95% confidence interval 2.6-5.5%) in their mothers after taking into account the sampling design and the weight of each sample. The children's HBsAg positivity was positively associated with maternal infection and being born in a non-health facility, while the maternal infection status was not associated with any background characteristic. CONCLUSIONS: Lao PDR has a relatively lower HBsAg prevalence in the general population compared to surrounding countries. To ensure comparability to other countries and to future data, rapid field tests are recommended for a nationwide prevalence survey.

Detecting Spread of Avian Influenza A(H7N9) Virus Beyond China.

During February 2013-March 2015, a total of 602 human cases of low pathogenic avian influenza A(H7N9) were reported; no autochthonous cases were reported outside mainland China. In contrast, since highly pathogenic avian influenza A(H5N1) reemerged during 2003 in China, 784 human cases in 16 countries and poultry outbreaks in 53 countries have been reported. Whether the absence of reported A(H7N9) outside mainland China represents lack of spread or lack of detection remains unclear. We compared epidemiologic and virologic features of A(H5N1) and A(H7N9) and used human and animal influenza surveillance data collected during April 2013-May 2014 from 4 Southeast Asia countries to assess the likelihood that A(H7N9) would have gone undetected during 2014. Surveillance in Vietnam and Cambodia detected human A(H5N1) cases; no A(H7N9) cases were detected in humans or poultry in Southeast Asia. Although we cannot rule out the possible spread of A(H7N9), substantial spread causing severe disease in humans is unlikely.

The Lao Experience in Deploying Influenza A(H1N1)pdm09 Vaccine: Lessons Made Relevant in Preparing for Present Day Pandemic Threats.

The Lao PDR, as did most countries of the Mekong Region, embarked on a pandemic vaccine initiative to counter the threat posed by influenza A(H1N1)pdm09. Overall, estimated vaccine coverage of the Lao population was 14%, with uptake in targeted health care workers and pregnant women 99% and 41%, respectively. Adverse Events Following Immunization accounted for only 6% of survey driven, reported vaccination experiences, with no severe consequences or deaths. Public acceptability of the vaccine campaign was high (98%). Challenges to vaccine deployment included: 1) no previous experience in fielding a seasonal influenza vaccine, 2) safety and efficacy concerns, and 3) late arrival of vaccine 10 months into the pandemic. The Lao success in surmounting these hurdles was in large measure attributed to the oversight assigned the National Immunization Program, and national sensitivities in responding to the avian influenza A(H5N1) crisis in the years leading up to the pandemic. The Lao "lessons learned" from pandemic vaccine deployment are made even more relevant four years on, given the many avian influenza strains circulating in the region, all with pandemic potential.

Influenza seasonality and vaccination timing in tropical and subtropical areas of southern and south-eastern Asia.

OBJECTIVE: To characterize influenza seasonality and identify the best time of the year for vaccination against influenza in tropical and subtropical countries of southern and south-eastern Asia that lie north of the equator. METHODS: Weekly influenza surveillance data for 2006 to 2011 were obtained from Bangladesh, Cambodia, India, Indonesia, the Lao People's Democratic Republic, Malaysia, the Philippines, Singapore, Thailand and Viet Nam. Weekly rates of influenza activity were based on the percentage of all nasopharyngeal samples collected during the year that tested positive for influenza virus or viral nucleic acid on any given week. Monthly positivity rates were then calculated to define annual peaks of influenza activity in each country and across countries. FINDINGS: Influenza activity peaked between June/July and October in seven countries, three of which showed a second peak in December to February. Countries closer to the equator had year-round circulation without discrete peaks. Viral types and subtypes varied from year to year but not across countries in a given year. The cumulative proportion of specimens that tested positive from June to November was > 60% in Bangladesh, Cambodia, India, the Lao People's Democratic Republic, the Philippines, Thailand and Viet Nam. Thus, these tropical and subtropical countries exhibited earlier influenza activity peaks than temperate climate countries north of the equator. CONCLUSION: Most southern and south-eastern Asian countries lying north of the equator should consider vaccinating against influenza from April to June; countries near the equator without a distinct peak in influenza activity can base vaccination timing on local factors.

National dengue surveillance in the Lao People's Democratic Republic, 2006-2012: epidemiological and laboratory findings.

Although dengue has been a public health problem for several decades in the Lao People's Democratic Republic, the magnitude of the disease burden and epidemiological trends remain poorly understood. We analysed national dengue surveillance and laboratory data from 2006 to 2012 by person, place and time. Between 2006 and 2012, the annual dengue notification rate ranged between 62 and 367 cases per 100 000 population with an apparent geographical expansion of transmission throughout the country in recent years and concurrent co-circulation of all four dengue virus subtypes. An electronic database, called Lao People's Democratic Republic Early Warning Alert and Response Network, was introduced in 2008 to provide automated early warning for outbreaks and epidemics. Village outbreaks continue to be notified primarily through event-based surveillance, whereas the weekly indicator-based system provides systematic assessment of annual epidemic cycles. The dengue case data indicate a high and increasing burden of disease. Efforts now need to focus on using available data to prompt more effective outbreak response and to guide the design and implementation of intervention strategies.

Chronic hepatitis B prevalence among children and mothers: results from a nationwide, population-based survey in Lao People's Democratic Republic.

BACKGROUND: Hepatitis B is regarded as a serious public health issue in Lao People's Democratic Republic (Lao PDR), a Southeast Asian country. However, disease epidemiology among the general population is not well known, and thus a nationwide cross-sectional survey for hepatitis B surface antigen (HBsAg) prevalence in children and their mothers was conducted. METHODS AND FINDINGS: We applied three-stage cluster sampling using probability proportionate to size. After randomly selecting child (5 to 9 years old) and mother (15 to 45 years old) pairs from the selected villages, questionnaires and HBsAg rapid tests were conducted. Data from 965 child and mother pairs were analyzed. Multivariate logistic regression analyses were used to investigate the independent association of individual background characteristics for the odds of being HBsAg positive. In total, 17 children and 27 mothers were HBsAg positive. HBsAg prevalence was estimated to be 1.7% (95% confidence interval: 0.8%-2.6%) in children, and 2.9% (95% confidence interval: 1.7%-4.2%) in their mothers after taking sampling design and weight of each sample into account. Mother's infection status was positively associated with HBsAg positivity in children (p<0.001), whereas other potential risk factors, such as ethnicity, proximity to health centers, and history of surgery, were not. There were no significant associations between mother's HBsAg status and history of surgery, and other sociodemographic factors. CONCLUSIONS: Despite the slow implementation of the hepatitis B vaccination program, HBsAg prevalence among children and their mothers was not high in Lao PDR compared to reports from neighboring countries. The reasons for the differences in prevalence among these countries are unclear. We recommend that prevalence surveys be conducted in populations born before and after the implementation of a hepatitis B vaccination program to better understand the epidemiology of hepatitis B.

An epidemic of dengue-1 in a remote village in rural Laos.

In the Lao PDR (Laos), urban dengue is an increasingly recognised public health problem. We describe a dengue-1 virus outbreak in a rural northwestern Lao forest village during the cool season of 2008. The isolated strain was genotypically "endemic" and not "sylvatic," belonging to the genotype 1, Asia 3 clade. Phylogenetic analyses of 37 other dengue-1 sequences from diverse areas of Laos between 2007 and 2010 showed that the geographic distribution of some strains remained focal overtime while others were dispersed throughout the country. Evidence that dengue viruses have broad circulation in the region, crossing country borders, was also obtained. Whether the outbreak arose from dengue importation from an urban centre into a dengue-naïve community or crossed into the village from a forest cycle is unknown. More epidemiological and entomological investigations are required to understand dengue epidemiology and the importance of rural and forest dengue dynamics in Laos.

Respiratory virus infections in hospitalized children and adults in Lao PDR.

BACKGROUND: Acute respiratory infections are an important cause of morbidity and mortality worldwide, with a major burden of disease in developing countries. The relative contribution of viruses in acute lower respiratory infections (ALRI) is, however, poorly documented in Lao PDR. OBJECTIVE: The objective of this study is to investigate the etiology of ALRI in patients of all ages in two hospitals of Laos. METHODS: Multiplex PCR/RT-PCR methods were used to target 18 major common respiratory viruses. Between August 2009 and October 2010, samples from 292 patients presenting with ALRI were collected. RESULTS AND CONCLUSION: Viruses were detected in 162 (55%) samples. In 48% (140/292) of the total ALRI cases, a single virus was detected while coinfections were observed in 8% (22/292) of the samples. The most frequent viruses were rhinovirus/enterovirus (35%), human respiratory syncytial virus (26%), and influenza viruses (13%). Parainfluenza viruses were detected in 9%, adenovirus in 6%, human metapneumovirus in 4%, coronaviruses (229E, NL63, OC43, HKU1) in 4%, and bocavirus in 3% of ALRI specimens. Most viral infections occurred in patients below 5 years of age. The distribution of viruses varied according to age-groups. No significant correlation was observed between the severity of the disease and the age of patients or the virus species. This study provides the description of viral etiology among patients presenting with ALRI in Lao PDR. Additional investigations are required to better understand the clinical role of the different viruses and their seasonality in Laos.

Epidemiological and virological characteristics of seasonal and pandemic influenza in Lao PDR, 2008-2010.

BACKGROUND: Information on influenza virology and epidemiology from Lao PDR is limited and the seasonal patterns of influenza have not been previously described. OBJECTIVES: To describe epidemiological and virologic characteristics of influenza in Lao PDR to recommend public health interventions, including improvements in surveillance and response. PATIENTS/METHODS: We performed a descriptive analysis of samples taken from patients with influenza-like-illness (ILI) (fever >38°C with cough and/or sore throat) presenting at seven sentinel hospitals in three regions of Lao PDR, January 2008-December 2010. A nasopharyngeal (NP) swab or combined nasal with oropharyngeal swab was collected from patients with ILI. Samples were tested for influenza by either Luminex RVP, conventional reverse transcriptase PCR (RT-PCR) (January 2008-2009), or by real-time PCR (rRT-PCR) using US CDC reagents (February 2009 onward). RESULTS: Of 2346 samples tested from patients with ILI, 523 (22%) were positive for influenza. The median age of those positive was 12 years (range, <1-60 year). The percentage of samples that were influenza positive was similar over the 3 years (20-23%). Each year 3-4 types/subtypes cocirculated with differing predominant type/subtype. Influenza was detected year-round with the highest proportion of positive specimens in the 3rd and 4th quarter. CONCLUSIONS: Similar to other countries in the region, we found that influenza is present year-round and has a peak activity from July to December. Dominant types or subtypes vary by year. A large proportion of patients with ILI are not influenza positive. ILI surveillance is critical for weighing disease burden, both morbidity and mortality, against the costs of advancing influenza vaccine delivery strategy.

Capacity building in response to pandemic influenza threats: Lao PDR case study.

The Lao People's Democratic Republic (PDR) committed to pandemic detection and response preparations when faced with the threat of avian influenza. Since 2006, the National Center for Laboratory and Epidemiology of Lao PDR has developed credible laboratory, surveillance, and epidemiological (human) capacity and as a result was designated a World Health Organization National Influenza Center in 2010. The Lao PDR experience in building influenza capacities provides a case study of the considerable crossover effect of such investments to augment the capacity to combat emerging and re-emerging diseases other than influenza.

Seroprevalence of rubella and measles antibodies in Lao PDR.

BACKGROUND: Nowadays, measles and rubella epidemics still occur worldwide, particularly in developing countries. METHODS: The present study investigated seroprevalence of rubella and measles IgG antibodies by using a comercial enzyme immunoassay. The findings helped to understand the current status of these infections in school children in Vientiane Capital, Lao PDR. RESULTS: The seropositive rate for measles antibody was 97.6%. Most importantly, more than half of 411 healthy children (56.4%) were rubella seronegative and susceptible to acquired rubella infection. CONCLUSIONS: The results highlight a high proportion of rubella-seronegative children, suggesting that they are susceptible to rubella and face the risk of congenital rubella syndrome. It was the first study to report on seroprevalence of rubella in Vientiane Capital, Lao PDR.

Serological investigations of flavivirus prevalence in Khammouane Province, Lao People's Democratic Republic, 2007-2008.

A large-scale cross-sectional seroprevalence study of dengue (DEN) and Japanese encephalitis (JE) was conducted in Khammouane province, Lao PDR, as part of the initial baseline health impact assessment of the Nam Theun 2 hydroelectric dam construction project. Health surveys were performed between May 2007 and February 2008 with serum samples collected from healthy individuals involved in the resettlement program of 16 villages (total surveyed population 4,369). Hemagglutination inhibition assay using flavivirus antigens (DENV1, DENV3, and JEV) performed on 1,708 plasma specimens revealed 30.4% (519) cross-reactive positives, and 10% (172) and 1.3% (22) positives to JEV or DENV, respectively. Entomological surveys conducted during the rainy season of 2008 indicated the presence of competent flavivirus vectors (Culex vishnui group and Aedes albopictus), although Aedes aegypti was not found. Continued surveillance and investigation is warranted to assess the clinical disease burden of flaviviruses in this area that is undergoing rapid ecological and demographic change.

An early report from newly established laboratory-based influenza surveillance in Lao PDR.

BACKGROUND: Prior to 2007, little information was available about the burden of influenza in Laos. We report data from the first laboratory-based influenza surveillance system established in the Lao People's Democratic Republic. METHODS: Three hospitals in the capital city of Vientiane began surveillance for influenza-like illness (ILI) in outpatients in 2007 and expanded to include hospitalized pneumonia patients in 2008. Nasal/throat swab specimens were collected and tested for influenza and other respiratory viruses by multiplex ID-Tag respiratory viral panel (RVP) assay on a Luminex 100x MAP IS instrument (Qiagen, Singapore). RESULTS: During January 2007 to December 2008, 287 of 526 (54.6%) outpatients with ILI were positive for at least one respiratory virus. Influenza was most commonly identified, with 63 (12.0%) influenza A and 92 (17.5%) influenza B positive patients identified. In 2008, six of 79 (7.6%) hospitalized pneumonia patients were positive for influenza A and four (5.1%) were positive for influenza B. Children <5 years represented 19% of viral infections in outpatients and 38% of pneumonia inpatients. CONCLUSION: Our results provide the first documentation of influenza burden among patients with febrile respiratory illness and pneumonia requiring hospitalization in Laos. Implementing laboratory-based influenza surveillance requires substantial investments in infrastructure and training. However, continuing outbreaks of avian influenza A/H5N1 in poultry and emergence of the 2009 influenza A(H1N1) pandemic strain further underscore the importance of establishing and maintaining influenza surveillance in developing countries.