Objective: To document the evolution and optimization of the Zika virus (ZIKV) disease surveillance system in southern Viet Nam in 2016 and to describe the characteristics of the identified ZIKV-positive cases. Methods: We established a sentinel surveillance system to monitor ZIKV transmission in eight sites in eight provinces and expanded the system to 71 sites in 20 provinces in southern Viet Nam in 2016. Blood and urine samples from patients who met the case definition at the sentinel sites were tested for ZIKV using real-time reverse transcription polymerase chain reaction at the Pasteur Institute in Ho Chi Minh City (PI-HCMC). We conducted descriptive analysis and mapped the ZIKV-positive cases. Results: In 2016, 2190 specimens from 20 provinces in southern Viet Nam were tested for ZIKV at PI-HCMC; 626 (28.6%), 484 (22.1%), 35 (1.6%) and 1045 (47.7%) tests were conducted in the first, second, third and fourth quarters of the year, respectively. Of these tested specimens, 214 (9.8%) were ZIKV positive with 212 (99.1%) identified in the fourth quarter. In the fourth quarter, the highest positivity rate was those in age groups 30–39 years (30.0%) and 40–59 years (31.6%). Of the 214 ZIKV-positive patients, 210 (98.1%) presented with rash, 194 (90.7%) with fever, 149 (69.6%) with muscle pain, 123 (57.5%) with joint pain and 66 (30.8%) with conjunctivitis. Discussion The surveillance system for ZIKV disease underwent several phases of optimization in 2016, guided by the most up-to-date local data. Here we demonstrate an adaptable surveillance system that detected ZIKV-positive cases in southern Viet Nam.

Objective: To examine the multiplication efficiency Japanese encephalitis virus (JEV) genotype I (GI) and genotype III (GIII) of different cell lines which originated from human, porcine, mosquitoes in order to prove mechanism of JEV GI replacement JEV GIII since it emerging in nature recent decades. Methods: The mixture of GI and GIII JEV isolates was inoculated on human rhabdomyosarcoma (RD), pig kidney epithelial (PS) and Aedes albopictus C6/36 clone (C6/36) which originated from human, porcine and mosquitoes, respectively. Plaque assays were performed to calculate virus titer and real-time RT-PCR with GI and GIII specific primer sets to quantify the number of GI and GIII RNA copies. Results: The highest virus titer reached at the 3rd day of post infection when GI and GIII mixture was inoculated on RD and PS and that of C6/36 was at the 4th day. JEVs were amplified and maintained by C6/36 cells after 10 passages whereas that by RD and PS only limited within 8 and 6 passages, respectively. GI strain amplified and maintained more efficiently on C6/36 and PS but not RD, whereas GIII strain amplified and maintained more efficiently on RD. Conclusions: There is a correlation between the multiplication efficiency of GI and GIII JEV strains when these two genotype strains co-infected on different cell lines with the predominance of GI strains in C6/36 and PS and the limited detection of GI strains in RD cells proving a possible mechanism of shift JEV genotypes in nature recent decades since GI emerging.

Objective: To establish a surveillance in Dong Thap, at the border with Cambodia by assessing the presence of DENV serotypes and CHIKV among patients hospitalized at Dong Thap general hospital. Methods: Cross-sectional descriptive analysis was conducted on a cohort of 131 patients hospitalized with acute fever and symptoms compatible with dengue or chikungunya. The study was conducted from January 2012 to February 2013. The full clinical picture was established as well as serological and molecular detection. Serological analysis was sequentially performed on blood samples collected on admission and an average of seven days after admission. The detection of IgM antibody to DENV was performed by IgM capture ELISA and the detection of DENV and CHIKV RNA was done by reverse-transcription multiplex PCR. Results: 101 patients out of 131 (77%) were confirmed with dengue. All four dengue serotypes were detected with a predominance of DENV2 and DENV4. No chikungunya infection was detected although reported in neighboring Cambodia. A differential efficiency of serological dengue detection was observed. Efficiency was 29% upon admission and 53% after seven days on the same patients. 30 patients out of 131 (23%) were negative with both DENV and CHIKV. Conclusions: Dengue is at risk of being underestimated and chikungunya is not systematically detected. Changes in detection and surveillance procedures are therefore discussed to increase efficiency of dengue detection and continue the monitoring the emergence of CHIKV in Dong Thap province and in Vietnam.

In this report we describe a new approach in HIV sentinel surveillance that was piloted in Viet Nam in 2009 and is currently being rolled out in all provinces. It comprises a brief behavioural questionnaire added to the HIV sentinel surveillance surveys conducted routinely among people who inject drugs, female sex workers and men who have sex with men. Timely reporting of data from this system has resulted in improvements to HIV prevention efforts for most at-risk populations.

Objective: To record the human cases of dengue fever (DF) and investigate the Aedes mosquito species circulating during the Hanoi 2011 DF epidemics. Methods: 24 different outbreak points were recorded in 8 districts between August and December 2011. Results: 140 patients were hospitalized following dengue diagnostic with a predominance of males (59.3%) and the 15-34 age class. Only DENV-1 (11.27%) and DENV-2 (88.73%) serotypes were detected in human samples. Mosquito sampling performed in and around patients households revealed the predominance of Aedes aegypti (A. aegypti) (95.15%) versus Aedes albopictus (4.85%). Conclusions: There is a positive correlation between the population density of A. aegypti and the number of human cases and duration of outbreaks. This was not observed for Aedes albopictus. Three pools of A. aegypti were positive with dengue virus, two with DENV-1 and one with DENV-2.

Background: Recently Japanese Encephalitis (JE) virus type 1 has surfaced and is co-circulated with JE virus type 3 in the northern areas of Viet Nam, so a sensitivity of JE viral antigen genotype 3 to detect IgM is required. Objectives: To compare the sensitivity of JE viral antigen genotype 1 and 3 to detect IgM against the JE virus. Materials and method: 783 cerebrospinal fluid and serum samples from viral encephalitis cases from 1999-2005 were collected and examined by MAC-ELISA for JE viral antigen genotype 1 and 3. Results: The agreement on the diagnosis of these kinds of antigen was 99.7% and the sensitivity of JE viral antigen genotype 3 was higher than that of genotype 1. Thus, JE viral antigen genotype 3 could be considered as the selected antigen for JE diagnosis in Viet Nam. IgM titer determined by JE viral antigen genotype 1 was higher than that of genotype 3 in 2003 and 2005 and lower in 1999, 2000, 2001, 2002 and 2004. Conclusion: The dominant phenomenon of JE viral genotypes differing over the years might be due to the interaction of the virus and its vectors. Further study is required to clarify this observation.
Japanese Encephalitis ; antigen

Background: Mosquitoes and pigs play important roles in maintaining and increasing the Japanese Encephalitis (JE) virus in nature and which is then transmitted to humans. Thus, surveillance of the JE infection frequency in the pig population may predict the human JE cases. \r\n', u'Objectives: The study aimed to determine IgG antibody against the JE virus in the pig population in Hanam province \r\n', u'Subjects and methods: The study included 1791 pig serum samples collected from 3 districts of Hanam province from Apr 2006 to Mar 2007. GAC-ELISA technique was used to determine the JE virus infection in the swine population.\r\n', u'Results: The average positive rate in pig population was 34.9 % (626/1791); with the highest frequency occurring in the summer (37.7%- 84.0 %), co-incident with the JE season in Northern Vietnam. On the contrary, in winter JE case are rare, frequency of IgG antibody against JE virus in the swine population was low, ranging from 9.2% to 22.0.%. \r\n', u'Conclusions: These results have shown the ecologically close relationship between the amplification of the JE virus in the swine population, vector and JE cases in northern Vietnam. \r\n', u'
Japanese encephalitis ; pig population ; GAC-ELISA.

Background: Japanese Encephalitis (JE) virus is a leading cause of encephalitis in children with high mortality and complication. JE is a dangerous infectious disease via Culex mosquitoes. Objective: To identify the density of some mosquitoes and vectors that transmit JE virus in Ha Nam province. Subject and Method:There were 30.333 mosquitoes including 05 genera and 15 species which were collected at night from April 2006 to March 2007 in the following communes: Tanson (midland area) Hungcong (plain area) and Liemchinh (suburban area). Results: A total of 06 Culex species, the highest rate is Culex tritaeniohychus (41.20%); Culex annulus (15.56%); other Culex species occupied around 0.15% -2.49%. In these communes, the density of Culex tritaeniohychus was higher than Culex annulus. They were found to be active all year round, but the highest density of Culex tritaeniohychus and Culex annulus is 5.86 and 2.15, respectively in April and fall to 4.35% and 1.71% in July. The collected mosquitoes during April, May, June and July includes 133 and 28 Culex tritaeniohychus and Culex annulus pools, respectively; all were to be processed for the isolation of Japanese Encephalitis (JE) virus. There were 12 Culex tritaeniohychus and 2 Culex annulus pools which were collected in Tanson commune yielding positive. Conclusion: The results were confirmed that Culex tritaenioohychus and Culex annulus are the major vectors that transmit JE virus in Ha Nam, 2006 - 2007. All 14 JE virus strains were isolated from collected mosquito pools during April, May and June; whereas JE virus strain was not yet isolated from collected mosquito pool in July. Thus, in order to control JE disease, it is necessary to control JE vectors in April, May and June.
Vector ; Culex tritaenioohychus ; Japanese encephalitis ; Ha Nam

Background: Japanese Encephalitis (JE) is common in the plains and mountainous areas in Asia \u2013 Pacific. Japanese encephalitis vaccine shows effectiveness in protecting children from JE in some countries such as Japan and Korea. Objective: To evaluate the efficacy of Japanese Encephalitis (JE) vaccination in Thai Binh province during 2003-2007. Subject and Method: Prospective, retrospective and sero-epidemiological methods were carried out on 329 samples collected from viral encephalitis patients and tested by JE MAC-ELISA, the positive average was 41.6% (137/329). Result: It had dramatically dropped from 85.2% in 2003 to 8.5 % in 2007 related to the rate of JE vaccination for children from 1 to 5 years old increasing from 49 % in 2003 to 77 % in 2007. Most of JE confirmed cases were un-vaccinated. Conclusion: JE etiology cause viral encephalitis in children in Thai Binh province was reduced thanks to JE vaccination in EPI program for 1 to 5 year old children. But more than 96% (131/136) of viral encephalitis in 15 years old upward was unknown etiology, the need for further study of the etiology cause viral encephalitis in adults.
Japanese encephalitis ; virus encephalitis ; MAC-ELISA ; Vaccine

Background: IgM antibody capture ELISA (MAC-ELISA) technique has been widely applied for Japanese Encephalitis Virus (JEV) diagnosis. So far rare internationally commercial kits are available. Thus, the international evaluation of the kit is required as per the recommendation of the WHO. Objectives: To evaluate the quality of the IgM antibody capture ELISA diagnostic kit for JEV produced by the Vietnam National Institute of Hygiene and Epidemiology (NIHE). Subjects and method: In this study, NIID kit was used as control to check the kit from NIHE. Both NIHE and NIID kits were used to detect JEV IgM among 38 serum and 6 CFS samples, which belongs to 5 sample groups (JE patients group, dengue patients group, other viral encephalitis patients group, Tick Born Encephalitis (TBE) patient group and healthy JE vaccinated donors group). Results: The detection of JEV IgM by NIHE kit was concurrent with the NIID kit. There is no positive with the JE in the groups of Dengue patients, TBE, other virus encephalitis patients and JE vaccinated donors. Conclusion: MAC-ELISA kit of NIHE can be used for different diagnosis of JEV and Dengue virus (both viruses are in Flavivirus genus), as well as other viruses caused by encephalitis.
IgM antibody ; ELISA diagnostic kit ; Japanese encephalitis virus