Whole Genome Sequencing of Dengue Virus Serotype 2 from Two Clinical Isolates and Serological Profile of Dengue in the 2015-2016 Nepal Outbreak.

Dengue virus (DENV) is the cause of one of the most prevalent neglected tropical diseases, and up to half of the world's population is at risk for infection. Recent results from clinical trials have shown that DENV vaccination can induce the development of severe dengue disease and/or prolong hospitalization after natural infection in certain naive populations. Thus, it is crucial that vaccine development takes into account the history of DENV exposure in the targeted population. In Nepal, DENV infection was first documented in 2004, and despite the increasing prevalence of DENV infection, the population remains relatively naive. However, it is not known which of the four DENV serotypes circulate in Nepal or whether there is evidence of repeated exposure to DENV in the Nepali population. To address this, we studied 112 patients who presented with symptomology suspicious for DENV infection at clinics throughout Nepal during late 2015 and early 2016. Of the 112 patients examined, 39 showed serological and/or genetic evidence of primary or secondary DENV infection: 30 were positive for DENV exposure by IgM/IgG ELISA, two by real-time reverse-transcription PCR (RT-PCR), and seven by both methods. Dengue virus 1-3, but not DENV4, serotypes were detected by RT-PCR. Whole genome sequencing of two DENV2 strains isolated from patients with primary and secondary infections suggests that DENV was introduced into Nepal through India, with which it shares a porous border. Further study is needed to better define the DENV epidemic in Nepal, a country with limited scientific resources and infrastructure.

Emergence of dengue in Nepal.

Dengue virus is a major health problem in Nepal. The endogenous dengue appeared in 2006 in the country with reported outbreaks in 2010, 2013 and 2016. Eleven years vertical data show there were sporadic cases in all the years and mostly adults between 25 and 40 years of age were infected with dengue virus. Compared with primary infections, secondary infections were observed in relatively larger numbers during the period of 2008-2016. Most of the cases had symptoms of dengue fever; while 7 and 19 cases demonstrated dengue hemorrhagic fever/dengue shock syndrome in 2010 and 2013 respectively. The proportion of dengue hemorrhagic fever amongst all cases of dengue fever was 2.5:4.7% in 2010 and 2013. We found there is shift of serotype from dengue virus serotype-1 (DENV-1) in 2010, DENV-2 in 2013 and DENV-1 in 2016. We feel there is urgent need for better community, hospital and laboratory based surveillance system capable of monitoring the circulating dengue virus (DENV) serotypes in different districts of Nepal. With improvement in surveillance system and efficient management of cases, the case fatality rate due to severe dengue can be reduced.

Dengue periodic outbreaks and epidemiological trends in Nepal.

Dengue is a global health problem and expansion of its endemics towards new territories in the hilly regions in Nepal is a serious concern. It appeared as a new disease in Nepal in 2004 from Japanese traveler with sporadic cases every year and massive outbreaks in 2010, 2013 and 2016. The serotype was responsible for outbreak in particular year was dengue virus serotype-1 (DENV-1) in 2010, 2016; and DENV-2 in 2013. Nepal lacks basic health related infrastructure in rural areas and does not have a stringent health care policy. With severances of epidemic like dengue, a new surveillance or an upgrading of existing one are direly needed to better challenge the possible outbreaks. This review paper aims to explain the dengue trend in last one decade in Nepal and warrants concerted and timely public health interventions to minimize the deleterious effects of the disease.

Dengue virus infection in a French traveller to the hilly region of Nepal in 2015: a case report.

BACKGROUND: Dengue viral infections are known to pose a significant risk during travel to tropical regions, but it is surprising to find dengue transmission in the hilly region of Nepal, which is over 1800mtr above sea level. CASE PRESENTATION: A 43-year-old Caucasian female traveler from France presented with fever and abdominal pain following a diarrheal illness while visiting the central hilly region of Nepal. Over the course of 9 days, she developed fever, body aches, and joint pain, with hemorrhagic manifestation. She was hospitalized in India and treated with supportive care, with daily monitoring of her platelets. An assessment by enzyme-linked immunosorbent assay showed that she was positive for dengue non-structural protein 1. Upon her return to France, dengue virus was confirmed by reverse transcriptase-polymerase chain reaction. CONCLUSION: The district where this dengue case was reported is in the hilly region of Nepal, neighboring the capital city Kathmandu. To the best of our knowledge, there has previously been no dengue cases reported from the district. This study is important because it aims to establish a potential region of dengue virus circulation not only in the tropics, but also in the subtropics as well, which in Nepal may exceed elevations of 1800mtr. This recent case report has raised alarm among concerned health personnel, researchers, and organizations that this infectious disease is now on the way to becoming established in a temperate climate.

Re-emergence of dengue virus serotype 2 strains in the 2013 outbreak in Nepal.

BACKGROUND & OBJECTIVES: Epidemiological interventions and mosquito control are the available measures for dengue control. The former approach uses serotype and genetic information on the circulating virus strains. Dengue has been frequently reported from Nepal, but this information is mostly lacking. The present study was done to generate a comprehensive clinical and virological picture of a dengue outbreak in Nepal during 2013. METHODS: A hospital-based study involving patients from five districts of Nepal was carried out. Demographic information, clinical details and dengue serological status were obtained. Viral RNA was characterized at the molecular level by reverse-transcription polymerase chain reaction (RT-PCR), nucleotide sequencing and phylogenetic analysis. RESULTS: From among the 2340 laboratory-confirmed dengue cases during the study period, 198 patients consented for the study. Clinically they had fever (100%), headache (59.1%), rashes (18.2%), retro-orbital pain (30.3%), vomiting (15.1%), joint pain (28.8%) and thrombocytopenia (74.3%). Fifteen (7.5%) of them had mucosal bleeding manifestations, and the rest were uncomplicated dengue fever. The patients were mostly adults with a mean age of 45.75 ± 38.61 yr. Of the 52 acute serum samples tested, 15 were positive in RT-PCR. The causative virus was identified as DENV serotype 2 belonging to the Cosmopolitan genotype. INTERPRETATIONS & CONCLUSIONS: We report here the involvement of DENV serotype 2 in an outbreak in Nepal in 2013. Earlier outbreaks in the region in 2010 were attributed to serotype 1 virus. As serotype shifts are frequently associated with secondary infections and severe disease, there is a need for enhancing surveillance especially in the monsoon and post-monsoon periods to prevent large-scale, severe dengue outbreaks in the region.

Brain perfusion single photon emission computed tomography abnormalities in patients with minimal hepatic encephalopathy.

BACKGROUND: Minimal hepatic encephalopathy (MHE) is the mildest form of hepatic encephalopathy (HE). Minimal hepatic encephalopathy patients do not demonstrate clinically overt symptoms of HE but present with abnormal neuropsychological and/or neurophysiological tests indicative of cerebral dysfunction. This study was performed in such patients to identify regions of abnormal cerebral perfusion and to correlate regional cerebral blood flow (rCBF) changes with psychometric hepatic encephalopathy score (PHES), Child-Turcotte-Pugh's score (CTP), and model for end-stage liver disease (MELD) score. We also compared abnormal patterns of rCBF in cirrhotic patients of alcoholic etiology with non-alcoholic etiology. METHODS: This prospective study was performed to evaluate rCBF in 50 cirrhotic patients and 13 controls using technetium-99m ethyl cysteinate dimer (Tc-99m ECD) brain single photon emission computed tomography. All the patients underwent a battery of psychometry tests, PHES. Minimal hepatic encephalopathy was diagnosed if PHES was ≤-5. The rCBF changes were evaluated using region of interest (ROI) based semi-quantitative method of region/cerebellum and region/cortex ratios in 16 regions of the brain. RESULTS: Cirrhotic patients with MHE showed impaired perfusion in the superior prefrontal cortex and increased perfusion in the thalamus, brain-stem, medial temporal cortex, and the hippocampus when compared with the controls. Cerebral perfusion in superior prefrontal cortex correlated negatively with MELD score (r=-0.323, P=0.022). We found significant positive correlation between PHES score and rCBF values in the left superior prefrontal cortex (r=0.385, P=0.006). Cirrhotic patients with alcohol etiology showed significantly decreased rCBF in right inferior prefrontal cortex, right superior prefrontal cortex, and the anterior cingulate cortex while increased rCBF was noted in the right medial temporal cortex and hippocampus. CONCLUSION: Our results suggest that alterations in cognition in cirrhotic patients with MHE may be associated with impaired abnormalities of rCBF.

Diagnosis and prognostic significance of minimal hepatic encephalopathy in patients with cirrhosis of liver.

BACKGROUND AND AIMS: Minimal hepatic encephalopathy is the mildest form of the spectrum of hepatic encephalopathy (HE) that impairs health-related quality of life. We assessed (1) the usefulness of psychometric hepatic encephalopathy score and critical flicker frequency for the diagnosis of minimal hepatic encephalopathy, and (2) prognostic significance of minimal hepatic encephalopathy. METHODS: One hundred patients with liver cirrhosis without overt HE were subjected to psychometric hepatic encephalopathy score and critical flicker frequency evaluation. Eighty-three age- and sex-matched healthy volunteers served as controls. Minimal hepatic encephalopathy was diagnosed when the psychometric hepatic encephalopathy score was or=8 [hazard ratio 2.466 (95% CI, 1.010-6.023)] predicted poor survival. CONCLUSIONS: Psychometric hepatic encephalopathy score is a useful tool for the diagnosis of minimal hepatic encephalopathy in an outpatient setting. Both psychometric hepatic encephalopathy score and Child-Turcotte-Pugh score have prognostic value on survival.