Australia as a global sink for the genetic diversity of avian influenza A virus.

Most of our understanding of the ecology and evolution of avian influenza A virus (AIV) in wild birds is derived from studies conducted in the northern hemisphere on waterfowl, with a substantial bias towards dabbling ducks. However, relevant environmental conditions and patterns of avian migration and reproduction are substantially different in the southern hemisphere. Through the sequencing and analysis of 333 unique AIV genomes collected from wild birds collected over 15 years we show that Australia is a global sink for AIV diversity and not integrally linked with the Eurasian gene pool. Rather, AIV are infrequently introduced to Australia, followed by decades of isolated circulation and eventual extinction. The number of co-circulating viral lineages varies per subtype. AIV haemagglutinin (HA) subtypes that are rarely identified at duck-centric study sites (H8-12) had more detected introductions and contemporary co-circulating lineages in Australia. Combined with a lack of duck migration beyond the Australian-Papuan region, these findings suggest introductions by long-distance migratory shorebirds. In addition, on the available data we found no evidence of directional or consistent patterns in virus movement across the Australian continent. This feature corresponds to patterns of bird movement, whereby waterfowl have nomadic and erratic rainfall-dependant distributions rather than consistent intra-continental migratory routes. Finally, we detected high levels of virus gene segment reassortment, with a high diversity of AIV genome constellations across years and locations. These data, in addition to those from other studies in Africa and South America, clearly show that patterns of AIV dynamics in the Southern Hemisphere are distinct from those in the temperate north.

Engagement Along the PrEP Care Continuum Among Men Who Have Sex with Men in China: A Systematic Review and Meta-analysis.

Pre-exposure prophylaxis (PrEP), including daily oral, on-demand, and long-acting injectable (LAI), is a promising HIV prevention intervention for men who have sex with men (MSM). We conducted a systematic review on engagement with the PrEP continuum among MSM in China. A total of 756 studies were initially identified and 36 studies were included (N = 26,021). In the 20 studies (N = 13,886) examining PrEP awareness, 32.4% (95% CI: 25.1-40.7) of MSM were aware of PrEP. In the 25 studies (N = 18,587) examining willingness, 54.5% (95% CI: 41.9-66.5) MSM indicated they were willing to use PrEP. The pooled prevalence of PrEP uptake from 9 studies (N = 6,575) was 4.9% (95% CI: 1.4-15.8%), while pooled estimates of adequate adherence from five studies (N = 2,344) among MSM on PrEP was 40.7% (95% CI: 20.0-65.2%). Subgroup analyses suggested studies conducted after 2015 (versus before) tended to report higher awareness and uptake. Awareness was highest for daily oral PrEP, followed by on-demand, and LAI PrEP; willingness to use was highest for LAI PrEP. The operationalization of willingness and adherence constructs varied across studies and complicated the interpretation of pooled estimates. This review revealed gaps in the PrEP care continuum among MSM in China, with relatively low awareness and uptake (in contrast to willingness and adherence) as the major potential barriers to widespread implementation and the need for a unified approach to defining and measuring PrEP outcomes.

Depression as a mediator between intimate partner violence (IPV) and CD4 cell count among men who have sex with men (MSM) living with HIV in China.

Intimate partner violence (IPV) is associated with adverse mental and physical outcomes among men who have sex with men (MSM) living with HIV. Few studies focus on psychological IPV, such as verbal threats. This study examined the associations between different forms of IPV and depression and CD4+ cell count, with depression as a mediator for the association between IPV and CD4+ cell count. Data for these analyses were derived from a larger cross-sectional study on HIV-HCV co-infection among MSM in Shanghai, China (N = 1623). We estimated the average causal mediation effects (ACME) and average direct effects (ADE) through three steps. About 16% of participants experienced IPV, with forced sex (7%), verbal threats (5%), and thrown objects (4%) being most common. Verbal threats showed the strongest link with depression and low CD4+ cell count. Depression fully mediated the relationship between verbal abuse and low CD4+ cell count, suggesting it as a potential pathway between psychological IPV and poorer HIV-related health outcomes. More research on psychological IPV is warranted to examine its health impacts. Mental health could be a potential focus of intervention to enhance HIV-related health outcomes among MSM with IPV experience.

Health-care access and utilization among HIV-infected men who have sex with men in two Chinese municipalities with or without lockdown amidst early COVID-19 pandemic.

City lockdown is critical to successfully contain the COVID-19 pandemic. The impact of lockdown and COVID-19 pandemic on healthcare among vulnerable population has yet to be explicated. A cross-sectional survey was conducted among HIV-infected men who have sex with men (MSM) in Wuhan with city lockdown and Shanghai without lockdown, and healthcare interruptions were evaluated and compared. A logistic regression analysis was employed to examine associates of HIV-related healthcare interruptions and compromised mental health. Compared to participants in Shanghai (N = 440), HIV-infected MSM in Wuhan (N = 503) had significantly higher proportion of untimely availability of antiretroviral drugs (ARVs) (20.6% vs. 8.4%), obtaining ARVs from outside institutions (29.1% vs. 8.1%), postponed non-AIDS treatment (6.4% vs. 2.8%) and untimely follow-up appointments (33.4% vs. 14.5%). HIV-related healthcare interruptions were positively associated with lockdown (OR = 4.89, 95% CI: 3.49-6.85) and non-local residence (OR = 1.91, 95% CI: 1.37-2.64). Compromised mental health, including insomnia and generalized anxiety disorders, was associated with non-local residence (OR = 1.35, 95% CI: 1.01-1.81) and healthcare interruptions (OR = 1.34, 95% CI: 1.01-1.79). HIV-infected MSM are vulnerable to healthcare interruptions and mental health problems during the COVID-19 pandemic, underscoring the need for tailored intervention strategies to minimize deleterious health consequences.

The Cumulative Influence of Perceived Discrimination, Stress, and Coping Responses on Symptoms of Depression Among Young African American Mothers.

BACKGROUND: African American women have an elevated risk for experiencing depressive symptoms, and discrimination, stress, and coping contribute to symptoms of depression. AIMS: We aimed to examine the associations between discrimination, stress, and coping on symptoms of depression among young African American mothers. METHODS: In this retrospective study, we utilized a hierarchical linear regression to explore the effects of perceived racial discrimination, stress, and general and discrimination-related coping responses on depressive symptoms in a sample of African American mothers (N = 250). The data were drawn from the Intergenerational Impact of Genetic and Psychological Factors on Blood Pressure study (InterGEN), a study conducted between 2014 and 2019 and based in Connecticut. RESULTS: After accounting for maternal age, level of education, and income, greater perceived racial discrimination (p = .03), higher levels of stress (p < .001), greater engagement in avoidance coping (p < .001), and use of passive coping responses to discrimination (p = .04) were uniquely associated with increased depressive symptoms. Other forms of coping, specifically, problem-solving and support seeking, did not appear to influence depressive symptoms in this sample. CONCLUSION: The findings highlight the negative impact of discrimination, heightened stress, and maladaptive coping on the emotional health of young African American mothers.

Intimate Partner Violence Among Chinese Transwomen: Associations With Sexual Risk Behaviors and HIV Testing.

BACKGROUND: Intimate partner violence (IPV) is an emerging risk factor for HIV infection. Given the high vulnerability of and limited research on transwomen in China, we described IPV, sexual risk behaviors, HIV, and sexually transmitted infection (STIs) testing rates and results, and investigated the pathways that link IPV to HIV infection among this population. METHODS: We conducted a cross-sectional survey and collected blood samples for HIV and syphilis testing among transwomen in Shanghai, China (n = 199). With logistic regression, we examined sexual risk behaviors and HIV/STI testing history among participants with and without IPV experience. RESULTS: More than half of the respondents reported IPV (57.3%), and the prevalence of unprotected sex ranged from 51.9% (with sex workers) to 87.8% (oral sex); 85.9% had ever tested for HIV and 49.3% for other STIs. Self-reported positivity results were as follows: HIV (2.3%), herpes simplex virus type 2 (8.3%), gonorrhea (18.8%), and syphilis (17.8%). Laboratory-confirmed positivity values were 5.0% for HIV and 6.5% for syphilis. Respondents with a history of IPV were significantly less likely to report HIV testing in the past 12 months (adjusted odds ratio, 0.20; 95% confidence interval, 0.10-0.38). CONCLUSIONS: Transwomen self-reported a high prevalence of IPV, which was related to a lower probability of HIV testing. The prevalence of HIV and other STIs was lower than reported in previous studies of Chinese transwomen, whereas the HIV/STI testing rates were higher. Findings suggest transwomen in China are at risk for IPV and need enhanced HIV prevention services to promote HIV testing in an IPV setting.

Pandemic potential of highly pathogenic avian influenza clade 2.3.4.4 A(H5) viruses.

The panzootic caused by A/goose/Guangdong/1/96-lineage highly pathogenic avian influenza (HPAI) A(H5) viruses has occurred in multiple waves since 1996. From 2013 onwards, clade 2.3.4.4 viruses of subtypes A(H5N2), A(H5N6), and A(H5N8) emerged to cause panzootic waves of unprecedented magnitude among avian species accompanied by severe losses to the poultry industry around the world. Clade 2.3.4.4 A(H5) viruses have expanded in distinct geographical and evolutionary pathways likely via long distance migratory bird dispersal onto several continents and by poultry trade among neighboring countries. Coupled with regional circulation, the viruses have evolved further by reassorting with local viruses. As of February 2019, there have been 23 cases of humans infected with clade 2.3.4.4 H5N6 viruses, 16 (70%) of which had fatal outcomes. To date, no HPAI A(H5) virus has caused sustainable human-to-human transmission. However, due to the lack of population immunity in humans and ongoing evolution of the virus, there is a continuing risk that clade 2.3.4.4 A(H5) viruses could cause an influenza pandemic if the ability to transmit efficiently among humans was gained. Therefore, multisectoral collaborations among the animal, environmental, and public health sectors are essential to conduct risk assessments and develop countermeasures to prevent disease and to control spread. In this article, we describe an assessment of the likelihood of clade 2.3.4.4 A(H5) viruses gaining human-to-human transmissibility and impact on human health should such human-to-human transmission occur. This structured analysis assessed properties of the virus, attributes of the human population, and ecology and epidemiology of these viruses in animal hosts.

Effectiveness of an Automated Error Checking and Feedback System to Improve Text Message Reporting for Disease Surveillance in Viet Nam.

Purpose: Viet Nam currently relies on a manual paper-based system to track and monitor 28 major infectious diseases. This inefficient system takes 2 or more months to complete. Method: We designed and pilot tested the use of text messaging to report certain infectious disease symptoms in rural areas of northern Viet Nam. The project was divided into three 6-month phases carried out in two provinces. The current analysis focuses on the implementation of a two-way feedback system between Phases II and III, which aims at (1) evaluating whether this system improves efficiency by determining the number of correctly (vs. incorrectly) formatted text messages; (2) assessing this system's influence on accuracy by comparing text messages with their respective official paper-based documented forms; and (3) determining whether the amount of information required to report through text messages influences the efficiency and accuracy of the text messages. Results: Between Phases II and III, results revealed a significant improvement in correctly formatted texts in comparison to incorrectly formatted texts. As the number of fields required to report increased, the number of correctly formatted texts (efficiency) as well as the number of matched text messages (accuracy) decreased. Conclusion: Our research demonstrates that an automated error bidirectional feedback system can significantly improve both the efficiency and accuracy of a Short Message Service-based method for disease surveillance. Also, our data may suggest that two-way communication has better engaged health care staff to follow reporting protocols as well as to maintain accuracy from their clinic's own data.

Associations between social determinants of health, perceived discrimination, and body mass index on symptoms of depression among young African American mothers.

PURPOSE: The association between symptoms of depression and risks for cardiovascular disease (CVD) remains equivocal for African American (AA) mothers. We examined the association between social determinants of health (perceived discrimination), and cardiovascular risk (BMI) on symptoms of depression in a sample of young AA mothers. METHODS: Secondary data from 219 adult AA mothers between the ages of 21 and 46 with an average BMI of 29.8 and yearly family income of $14,999 were analyzed using a latent growth model that evaluated four time points to assess changes in symptoms of depression. RESULTS: Initial BMI was significantly associated with initial symptoms of depression (b = 0.12, p = .019). Perceived discrimination (unfair treatment) was associated with higher initial symptoms of depression (b = 1.14, p = .017). CONCLUSION: The findings suggest that elevated BMI and perceived discrimination are associated with higher reported symptoms of depression among young, socioeconomically disadvantaged AA mothers. These results advance the scientific understanding of young AA mothers' risk for symptoms of depression and CVD by elucidating the impact of perceived discrimination and social experiences on mental health. Further studies of SDoH and CVD risk factors and perceived racism and depression are needed to shed light on the long-term mental health impact on AA mothers and their children.

Locally Acquired Human Infection with Swine-Origin Influenza A(H3N2) Variant Virus, Australia, 2018.

In 2018, a 15-year-old female adolescent in Australia was infected with swine influenza A(H3N2) variant virus. The virus contained hemagglutinin and neuraminidase genes derived from 1990s-like human seasonal viruses and internal protein genes from influenza A(H1N1)pdm09 virus, highlighting the potential risk that swine influenza A virus poses to human health in Australia.

Actives, passives and power: heteronormative gender norms and their implications for intimate partner violence among men who have sex with men in Tajikistan.

Global research reveals that gender-role norms have a profound effect on socio- and psycho-sexual expression (e.g. sexual positioning) among gay and bisexual men, which in turn may affect mental health and sexual risks. However, little is known about these factors among gay and bisexual men in Muslim-majority countries such as Tajikistan. Using a combination of in-depth individual interviews and focus-group assessments, this exploratory, qualitative study examined how gender roles might function as a social determinant for the practice of sexual positioning, which in turn may influence intimate partner violence (IPV), sexual risk and relational power. Results suggest that being the 'active' partner in sexual relationships closely aligns with the construct of hegemonic masculinity, affording actives more power in male-male relationships which may in some cases result in IPV directed against 'passives'. Despite this imbalance, passives also hold power in some cases, such as easier access to public resources such as the police and gay and bisexual focused services. Further research should examine gender norms and sexual positioning relative to IPV and sexual risks among Muslim men in Tajikistan.

Serologic Evidence of Exposure to Highly Pathogenic Avian Influenza H5 Viruses in Migratory Shorebirds, Australia.

Highly pathogenic avian influenza (HPAI) H5Nx viruses of the goose/Guangdong/96 lineage continue to cause outbreaks in poultry and wild birds globally. Shorebirds, known reservoirs of avian influenza viruses, migrate from Siberia to Australia along the East-Asian-Australasian Flyway. We examined whether migrating shorebirds spending nonbreeding seasons in Australia were exposed to HPAI H5 viruses. We compared those findings with those for a resident duck species. We screened >1,500 blood samples for nucleoprotein antibodies and tested positive samples for specific antibodies against 7 HPAI H5 virus antigens and 2 low pathogenicity avian influenza H5 virus antigens. We demonstrated the presence of hemagglutinin inhibitory antibodies against HPAI H5 virus clade 2.3.4.4 in the red-necked stint (Calidris ruficolis). We did not find hemagglutinin inhibitory antibodies in resident Pacific black ducks (Anas superciliosa). Our study highlights the potential role of long-distance migratory shorebirds in intercontinental spread of HPAI H5 viruses.

Divergent Human-Origin Influenza Viruses Detected in Australian Swine Populations.

Global swine populations infected with influenza A viruses pose a persistent pandemic risk. With the exception of a few countries, our understanding of the genetic diversity of swine influenza viruses is limited, hampering control measures and pandemic risk assessment. Here we report the genomic characteristics and evolutionary history of influenza A viruses isolated in Australia from 2012 to 2016 from two geographically isolated swine populations in the states of Queensland and Western Australia. Phylogenetic analysis with an expansive human and swine influenza virus data set comprising >40,000 sequences sampled globally revealed evidence of the pervasive introduction and long-term establishment of gene segments derived from several human influenza viruses of past seasons, including the H1N1/1977, H1N1/1995, H3N2/1968, and H3N2/2003, and the H1N1 2009 pandemic (H1N1pdm09) influenza A viruses, and a genotype that contained gene segments derived from the past three pandemics (1968, reemerged 1977, and 2009). Of the six human-derived gene lineages, only one, comprising two viruses isolated in Queensland during 2012, was closely related to swine viruses detected from other regions, indicating a previously undetected circulation of Australian swine lineages for approximately 3 to 44 years. Although the date of introduction of these lineages into Australian swine populations could not be accurately ascertained, we found evidence of sustained transmission of two lineages in swine from 2012 to 2016. The continued detection of human-origin influenza virus lineages in swine over several decades with little or unpredictable antigenic drift indicates that isolated swine populations can act as antigenic archives of human influenza viruses, raising the risk of reemergence in humans when sufficient susceptible populations arise.IMPORTANCE We describe the evolutionary origins and antigenic properties of influenza A viruses isolated from two separate Australian swine populations from 2012 to 2016, showing that these viruses are distinct from each other and from those isolated from swine globally. Whole-genome sequencing of virus isolates revealed a high genotypic diversity that had been generated exclusively through the introduction and establishment of human influenza viruses that circulated in past seasons. We detected six reassortants with gene segments derived from human H1N1/H1N1pdm09 and various human H3N2 viruses that circulated during various periods since 1968. We also found that these swine viruses were not related to swine viruses collected elsewhere, indicating independent circulation. The detection of unique lineages and genotypes in Australia suggests that isolated swine populations that are sufficiently large can sustain influenza virus for extensive periods; we show direct evidence of a sustained transmission for at least 4 years between 2012 and 2016.

Evolving HIV Epidemiology in Mainland China: 2009-2018.

PURPOSE OF REVIEW: This review is intended to provide an overview of the evolution of HIV epidemiology over the past decade in China. RECENT FINDINGS: We provided a succinct overall view of the epidemic, followed by surveillance data, profiles of key populations, HIV molecular epidemiology, and drug resistance, as well as survival in the age of antiretroviral therapy usage. For each topical issue, we first reviewed the latest empirical evidence, followed by a brief summary assessment. We briefly addressed the challenges and opportunities of the next generation of HIV control and prevention efforts in China. Notably, macro-social factors need to be integrated into the next generation of clinical and/or behavioral HIV research to inform disease progression and management, as well as control and prevention.

HIV and Aging in Mainland China: Implications for Control and Prevention Research.

PURPOSE OF REVIEW: The last 15 years have witnessed a dramatic change in HIV-related epidemiology amidst improvements in treatment and care in China. With proper treatment, HIV is now considered a chronic disease. As a consequence, many people living with HIV (PLWH) now present age-related comorbidities. We reviewed 13 topical issues concerning the epidemiology of aging with HIV in mainland China. RESULTS: Many of aging-related issues associated with the biological and physical aspects of living with HIV addressed in mainland China are consistent with the global literature. Greater prevalence of age-related comorbidities among PLWH was observed. Beyond biological mechanisms associated with HIV infection and traditional risk factors, other factors play a vital role in the aging process among PLWH. A stronger focus on screening, prevention, and management of non-HIV co-morbidities among PLWH is now warranted. Macro-social factors need to be integrated into next generation of clinical and/or behavioral HIV research to inform disease progression and management as well as prevention.

Higher Prevalence of Frailty Among a Sample of HIV-Infected Middle-aged and Older Chinese Adults Is Associated With Neurocognitive Impairment and Depressive Symptoms.

We investigated the prevalence and correlates of prefrailty/frailty, determined on the basis of the Fried criteria, in Chinese patients with and those without human immunodeficiency virus (HIV) infection. HIV-infected patients were more likely to be frail or prefrail than controls, and this association remained significant after adjustment for potential confounders (odds ratio, 3.79). After additional adjustment for neurocognitive impairment and depressive and insomnia symptoms, this association remained significant but attenuated (odds ratio, 2.16). In the HIV-infected group, these 3 variables were independently associated with prefrailty/frailty. These findings suggest that neurocognitive impairment and depressive and/or insomnia symptoms may account for a higher prevalence of prefrailty/frailty in HIV-infected patients but require further longitudinal investigation.

Estimating self-reported sex practices, drug use, depression, and intimate partner violence among MSM in China: a comparison of three recruitment methods.

Men who have sex with men (MSM) are the group at highest risk for HIV in China. Researchers have used various recruitment methods to reach this population hidden from the hetero-normative culture. To inform future recruitment strategies, we compared estimates of socio-demographic characteristics, HIV risk behaviors, depression, and intimate partner violence (IPV) across three samples of MSM and money boys in Shanghai, China. Data were collected from three community-based samples of MSM and money boys (n = 1352) recruited via respondent-driven sampling (RDS) (n = 404), community popular opinion leaders (CPOL) (n = 385), and Internet and venue-based sampling (VBS) (n = 546). Different recruitment methods generated samples with statistically significant differences among a number of socio-demographic characteristics, sexual behaviors, drug use, depression scores, and exposure to IPV. Specifically, RDS participants had lower education (p = .002), income levels (p < .001), and were more likely to report condomless sex with a woman (p < .001). CPOL participants were younger (p < .001), more likely to report lifetime condomless anal sex (p = .009), more than 10 male partners in the past 30 days (p < .001), and were less likely to experience violence by a male intimate partner (p = .001). VBS participants had lowest depression score (p = .005) and were more likely to report lifetime drug use (p = .003). Our findings reinforce that each recruitment method may reach a sub-group of MSM with a specific risk profile, so multiple methods may be needed to obtain a representative sample of MSM. Interventions may use specific recruitment methods to target certain segments of the MSM population.

Stigmatization of people who inject drugs (PWID) by pharmacists in Tajikistan: sociocultural context and implications for a pharmacy-based prevention approach.

BACKGROUND: Pharmacies are an important source of sterile syringes for people who inject drugs (PWID) in Tajikistan who are under high risk of HIV and hepatitis C virus. Accessibility of sterile syringes at pharmacies without prescription may depend on pharmacists' attitudes towards PWID. This qualitative inquiry examines meanings and processes of stigmatization of PWID among pharmacists and pharmacy students in Tajikistan. METHODS: We conducted semi-structured interviews with 19 pharmacists and 9 students (N = 28) in the cities of Dushanbe and Kulob, Tajikistan. The interview topics included personal attitudes towards drug use and PWID, encounters with PWID, awareness and beliefs related to drug dependence and HIV, and attitudes and practices related to providing syringes to PWID. Interview transcripts were analysed using thematic analysis methods. RESULTS: The main themes included the significance of religion in defining attitudes towards drug use, labelling of PWID, negative stereotypes (PWID are prone to crime, violence, and irrational aggression; inflict harm to families and society; are able to control drug use), emotions triggered by PWID (fear, sympathy) and discrimination against PWID (rejection, isolation, ostracism, limiting resources to PWID). The religious ban on drug use and pharmacists' moral and legal responsibility for the consequences of drug use were frequently mentioned as reasons for rejecting syringe sales. Still, many participants acknowledged the need for distributing syringes to PWID to prevent HIV. CONCLUSIONS: Stigma against PWID in Tajikistan plays an important role in shaping pharmacists' attitudes towards provision of services to this population. Local sociocultural context, in particular religious beliefs and social conservatism, may facilitate stigmatizing beliefs.

Reassortant highly pathogenic influenza A(H5N6) virus in Laos.

In March 2014, avian influenza in poultry in Laos was caused by an emergent influenza A(H5N6) virus. Genetic analysis indicated that the virus had originated from reassortment of influenza A(H5N1) clade 2.3.2.1b, variant clade 2.3.4, and influenza A(H6N6) viruses that circulate broadly in duck populations in southern and eastern China.

Detection of highly pathogenic zoonotic influenza virus H5N6 by reverse-transcriptase quantitative polymerase chain reaction.

BACKGROUND: Variant high pathogenicity avian influenza (HPAI) H5 viruses have recently emerged as a result of reassortment of the H5 haemagglutinin (HA) gene with different neuraminidase (NA) genes, including NA1, NA2, NA5, NA6 and NA8. These viruses form a newly proposed HA clade 2.3.4.4 (previously provisionally referred to as clade 2.3.4.6), and have been implicated in disease outbreaks in poultry in China, South Korea, Laos, Japan and Vietnam and a human fatality in China. There is real concern that this new clade may be wide spread and not readily identified using existing diagnostic algorithms. FINDINGS: Fluorescent probe based reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR) assays were developed to facilitate the identification of novel clade 2.3.4.4 viruses of H5N6 subtype emerging in Asia. Assays were aimed at the haemagglutinin (HA) gene for clade identification and at the NA gene to identify N6. The HA assay employing a minor groove binder (MGB) probe was able to detect and differentiate A/duck/Laos/XBY004/2014(H5N6) and related influenza A(H5N6) virus isolates belonging to the proposed clade 2.3.4.4 from other H5 HPAI viruses. In addition, an Eurasian N6 assay was able to differentiate N6 from other NA subtypes. CONCLUSIONS: Laos influenza A(H5N6) virus representative of proposed clade 2.3.4.4, was detected and differentiated from viruses in other H5N1 clades using a clade-specific HA RT-qPCR assay whereas the N6-NA subtype was determined by an Eurasian N6 RT-qPCR assay. Such a clade-specific assay would be of particular value for surveillance and in diagnostic laboratories where sequencing is not readily available.