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Background & objectives: Nipah virus (NiV) is a zoonotic paramyxovirus that causes fatal encephalitis in humans. Enzyme Linked Immunosorbent Assay (ELISA) is a safe, sensitive, specific, and affordable diagnostic tool that can be used during screening of large-scale epidemiological investigations. Development and evaluation of IgM and IgG ELISA for screening serum samples of NiV suspected cases would also help in planning public health interventions. Methods: An IgM capture (MAC) ELISA and an indirect IgG ELISA were developed using NiV antigen to detect IgM and IgG antibodies against NiV in human sera. The sensitivity, specificity, and cross- reactivity of the assays were evaluated using NiV IgM, IgG positive, negative human sera and measles, mumps, rubella, Crimean-Congo haemorrhagic fever, Kyasanur forest disease IgM, IgG positive sera, respectively. Results: The developed anti-NiV IgM and IgG ELISAs have shown specificity of 99.28 per cent and sensitivity of 100 per cent compared to reference test from Centers for Disease Control and Prevention, USA. Assays demonstrated negative predictive value of 100 per cent and positive predictive value as 90 and 93.94 per cent for anti-Nipah IgM ELISA and IgG ELISA respectively with test accuracy of 99.33 per cent. Interpretation & conclusions: Timely diagnosis of NiV is crucial for the management of cases, which could prevent further spread of infection in the community. IgM ELISA can be used as primary diagnostic tool followed by polymerase chain reaction. These assays have advantages of its applicability during outbreak investigations and surveillance activities at hospital or onsite laboratories with basic biosafety practices.
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Objective:To screen and identify H-2 d-restricted T cell epitopes in fusion (F) and attachment (G) glycoproteins of Nipah virus (NiV) in mice. Methods:The complete peptides (single peptide contains 15 amino acids, and 10 amino acids were repeated in the front and back peptides) derived from F and G antigens were mixed into peptide libraries. BALB/c mice were immunized with DNA vaccines expressing NiV F and G proteins alone and in combination. The full sequence peptide libraries of F and G antigens were mixed into peptide pools by matrix design, and spleen cells of immunized mice were collected and analyzed by IFN-γ ELISPOT assay to detect the dominant H-2 d-restricted epitope peptides. Results:Twelve dominant H-2 d-restricted peptides were screened from the F protein-specific peptide library and the 56th peptide produced the strongest reaction. Four dominant peptides were screened from the G protein-specific peptide library and the 72nd peptide produced the strongest reaction. Conclusions:In this study, 12 F antigen-specific and 4 G antigen-specific H-2 d restricted dominant T cell epitopes of NiV were screened and identified by IFN-γ ELISPOT, which could provide reference for immunological analysis of NiV and vaccine research.
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Introduction: The first Nipah Virus (NiV) outbreak occurred in India in the year 2001 at Siliguri. The second outbreak happened at Nadia in 2007. Nipah Virus exhibits neurological and pneumonic tropism with the predominant clinical presentation being encephalitis in humans. Material and Methods: The present study was a record based prospective study on 67 cases admitted with pyrexia of unknown origin in North Bengal Medical College during the period from 18.02.2001 to 30.02.2001 and a parallel study on epidemiological record carried out by PSM department also taken into account. All necessary investigations including autopsy examination, pathological, and microbiological study were done. Results: There was a clustering of cases around Bhaktinagar. There was a strong H/O Medinova Nursing Home Contact among the patients. 18 out of 20 cases were staff of that Nursing Home. Serum samples tested show NiV specific IgM and IgG in 9 out of 17 samples with one sample which was positive for IgG only suggesting past infection. The cases were admitted with predominant neurological symptoms (53.73% cases) but about 80% recovered with no residual neuro deficit. The natural reservoir of NiV is present in Bangladesh and in Northern India. Conclusion: When NiV infection is suspected, infection control practices must be strengthened to avoid an outbreak in a hospital setting. Here the present study is presenting the experience in the first outbreak of the Nipah virus in India at Siliguri for awareness of clinical personnel to control further outbreak at the very beginning.
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Nipah Virus is a recently emerging zoonotic virus with disease causing potential in both animals and humans. Nipah virus belongs to the family of paramyxovirida, genus Henipavirus along with Hendra virus. (1) The knowledge of human infection with Henipavirus was limited to a very small number of cases infected with Hendra virus in Australia during 1994-1999 which was responsible for deaths of two humans and seventeen horses. (2) Nipah virus was first identified and isolated in 1999 in Malaysia during an outbreak of febrile illness among pig farmers and people who were in close contacts with pigs. (3) In 2001, Nipah virus was identified as the causative agent of outbreak in Bangladesh. Since then number of outbreaks has been reported in various districts of Bangladesh. (4) In India, a total of three outbreaks of Nipah have been reported, latest being on 19th May 2018, from Kozhikode district of Kerala. (5) With a fatality rate of 58%, Nipah virus is primarily seen to cause encephalitis and severe respiratory distress. Despite of the severe pathogenicity and high pandemic potential there is no specific treatment for Nipah virus encephalitis except for supportive and symptomatic treatment.
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Nipah virus infection is a newly emerging zoonotic disease that spreads from animal to human. The fruit bats are the one of the carriers of Nipah virus which gets transmitted from bats to other animals mainly through body fluids. Nipah can cause asymptomatic infection to the acute respiratory syndrome and fatal encephalitis in human beings with a dreadful mortality rate of 70 per cent. This review article focused on role of bats in ecosystem and preventive measure to contain the virus by improving public health. In 2018 the outbreak of Nipah in Kerala state of India claimed 17 lives. It has indeed opened Pandora’s box exposing the role of general public, health department and social media and so on. The early detection and the well-oiled medical infrastructure in the state did wonders. Though social media have been accused of spreading misinformation leading to culling of bats, the people of Kerala responded with positive frame of mind. Prompt and synchronized efforts of all stakeholders like health workers, state government, scientists, researchers and central government led to timely and successful control of Nipah virus cases in Kerala.
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Infectious diseases remain as the major causes of human and animal morbidity and mortality leading to significant healthcare expenditure in India. The country has experienced the outbreaks and epidemics of many infectious diseases. However, enormous successes have been obtained against the control of major epidemic diseases, such as malaria, plague, leprosy and cholera, in the past. The country's vast terrains of extreme geo-climatic differences and uneven population distribution present unique patterns of distribution of viral diseases. Dynamic interplays of biological, socio-cultural and ecological factors, together with novel aspects of human-animal interphase, pose additional challenges with respect to the emergence of infectious diseases. The important challenges faced in the control and prevention of emerging and re-emerging infectious diseases range from understanding the impact of factors that are necessary for the emergence, to development of strengthened surveillance systems that can mitigate human suffering and death. In this article, the major emerging and re-emerging viral infections of public health importance have been reviewed that have already been included in the Integrated Disease Surveillance Programme.
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Nipah Virus (NiV) is a biological disaster and zoonotic pathogen which can be transmitted from animal to human beings. TheNiV was first identified in Malaysia in 1998. In India it was traced at Siliguri in 2001, followed by second outbreak in Nadiadistrict of West Bengal in 2007 and the present outbreak consumed more than 17 lives and affected many in Kerala, a southernIndian state which is globally known as “God’s own country”. The “all-time alert care” provided by the Kerala State HealthDepartment have earned applause for early detection of Nipah outbreak. The prompt action ensured containing the spread ofNipah outbreak and halting a major catastrophe, in spite of the best efforts the anxiety and panic was commonly reported amongthe communities. The current review is to explore the psychosocial perspectives of NiV and its impact in Kerala. Studies onNiV were collected from different online search engines, journals and newspapers. The review points out that there is a needto address psychosocial aspects of NiV along with pharmacological intervention to reduce vulnerability by enhancing bettercoping and resilience of individual, family and community.
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Abstracts@#This study aims to determine the importance of conserved GDN motif in domain III and GXGXG motif in domain VI in Nipah virus (NiV) L protein. Four mutated L genes produced in an earlier study were inserted individually into plasmid pCITE. Optimised transfection protocol was successful in transfecting these plasmids, two helper plasmids (coding for N and P protein), NiV minigenome containing chloramphenicol acetyltransferase (CAT) reporter gene and T7 promoter. Successful in vitro transcription/translation in the NiV minireplicon system was monitored by CAT expression. In conclusion, GXGXG motif was important in the NiV minireplicon system but change of GDN motif does not affect L protein.
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Objective To explore a common B- and T-cell epitope-based vaccine that can elicit an immune response against encephalitis causing genus Henipaviruses, Hendra virus (HeV) and Nipah virus (NiV). Methods Membrane proteins F, G and M of HeV and NiV were retrieved from the protein database and subjected to different bioinformatics tools to predict antigenic B-cell epitopes. Best B-cell epitopes were then analyzed to predict their T-cell antigenic potentiality. Antigenic B- and T-cell epitopes that shared maximum identity with HeV and NiV were selected. Stability of the selected epitopes was predicted. Finally, the selected epitopes were subjected to molecular docking simulation with HLA-DR to confirm their antigenic potentiality in silico. Results One epitope from G proteins, one from M proteins and none from F proteins were selected based on their antigenic potentiality. The epitope from the G proteins was stable whereas that from M was unstable. The M-epitope was made stable by adding flanking dipeptides. The 15-mer G-epitope (VDPLRVQWRNNSVIS) showed at least 66% identity with all NiV and HeV G protein sequences, while the 15-mer M-epitope (GKLEFRRNNAIAFKG) with the dipeptide flanking residues showed 73% identity with all NiV and HeV M protein sequences available in the database. Molecular docking simulation with most frequent MHC class-II (MHC II) and class-I (MHC I) molecules showed that these epitopes could bind within HLA binding grooves to elicit an immune response. Conclusions Data in our present study revealed the notion that the epitopes from G and M proteins might be the target for peptide-based subunit vaccine design against HeV and NiV. However, the biochemical analysis is necessary to experimentally validate the interaction of epitopes individually with the MHC molecules through elucidation of immunity induction.
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OBJECTIVE@#To explore a common B- and T-cell epitope-based vaccine that can elicit an immune response against encephalitis causing genus Henipaviruses, Hendra virus (HeV) and Nipah virus (NiV).@*METHODS@#Membrane proteins F, G and M of HeV and NiV were retrieved from the protein database and subjected to different bioinformatics tools to predict antigenic B-cell epitopes. Best B-cell epitopes were then analyzed to predict their T-cell antigenic potentiality. Antigenic B- and T-cell epitopes that shared maximum identity with HeV and NiV were selected. Stability of the selected epitopes was predicted. Finally, the selected epitopes were subjected to molecular docking simulation with HLA-DR to confirm their antigenic potentiality in silico.@*RESULTS@#One epitope from G proteins, one from M proteins and none from F proteins were selected based on their antigenic potentiality. The epitope from the G proteins was stable whereas that from M was unstable. The M-epitope was made stable by adding flanking dipeptides. The 15-mer G-epitope (VDPLRVQWRNNSVIS) showed at least 66% identity with all NiV and HeV G protein sequences, while the 15-mer M-epitope (GKLEFRRNNAIAFKG) with the dipeptide flanking residues showed 73% identity with all NiV and HeV M protein sequences available in the database. Molecular docking simulation with most frequent MHC class-II (MHC II) and class-I (MHC I) molecules showed that these epitopes could bind within HLA binding grooves to elicit an immune response.@*CONCLUSIONS@#Data in our present study revealed the notion that the epitopes from G and M proteins might be the target for peptide-based subunit vaccine design against HeV and NiV. However, the biochemical analysis is necessary to experimentally validate the interaction of epitopes individually with the MHC molecules through elucidation of immunity induction.
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Nipah virus (NiV) is a member of the genus Henipavirus of the family Paramyxoviridae, characterized by high pathogenicity and endemic in South Asia. It is classified as a Biosafety Level-4 (BSL-4) agent. The case-fatality varies from 40% to 70% depending on the severity of the disease and on the availability of adequate healthcare facilities. At present no antiviral drugs are available for NiV disease and the treatment is just supportive. Phylogenetic and evolutionary analyses can be used to help in understanding the epidemiology and the temporal origin of this virus. This review provides an overview of evolutionary studies performed on Nipah viruses circulating in different countries. Thirty phylogenetic studies have been published from 2000 to 2015 years, searching on pub-med using the key words ‘Nipah virus AND phylogeny’ and twenty-eight molecular epidemiological studies from 2006 to 2015 have been performed, typing the key words ‘Nipah virus AND molecular epidemiology’. Overall data from the published study demonstrated as phylogenetic and evolutionary analysis represent promising tools to evidence NiV epidemics, to study their origin and evolution and finally to act with effective preventive measure.
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Nipah virus (NiV) is a member of the genus Henipavirus of the family Paramyxoviridae, characterized by high pathogenicity and endemic in South Asia. It is classified as a Biosafety Level-4 (BSL-4) agent. The case-fatality varies from 40% to 70% depending on the severity of the disease and on the availability of adequate healthcare facilities. At present no antiviral drugs are available for NiV disease and the treatment is just supportive. Phylogenetic and evolutionary analyses can be used to help in understanding the epidemiology and the temporal origin of this virus. This review provides an overview of evolutionary studies performed on Nipah viruses circulating in different countries. Thirty phylogenetic studies have been published from 2000 to 2015 years, searching on pub-med using the key words 'Nipah virus AND phylogeny' and twenty-eight molecular epidemiological studies from 2006 to 2015 have been performed, typing the key words 'Nipah virus AND molecular epidemiology'. Overall data from the published study demonstrated as phylogenetic and evolutionary analysis represent promising tools to evidence NiV epidemics, to study their origin and evolution and finally to act with effective preventive measure.
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Between September 1998 to May 1999, Malaysia and Singapore were hit by an outbreak of fatal encephalitis caused by a novel virus from the paramyxovirus family. This virus was subsequently named as Nipah virus, after the Sungei Nipah village in Negeri Sembilan, where the virus was first isolated. The means of transmission was thought to be from bats-topigs and subsequently pigs-to-human. Since 2001, almost yearly outbreak of Nipah encephalitis has been reported from Bangladesh and West Bengal, India. These outbreaks were characterized by direct bats-to-human, and human-to-human spread of infection. Nipah virus shares many similar characteristics to Hendra virus, first isolated in an outbreak of respiratory illness involving horses in Australia in 1994. Because of their homology, a new genus called Henipavirus (Hendra + Nipah) was introduced. Henipavirus infection is a human disease manifesting most often as acute encephalitis (which may be relapsing or late-onset) or pneumonia, with a high mortality rate. Pteropus bats act as reservoir for the virus, which subsequently lead to human spread. Transmission may be from consumption of food contaminated by bats secretion, contact with infected animals, or human-to-human spread. With wide geographical distribution of Pteropus bats, Henipavirus infection has become an important emerging human infection with worldwide implication.
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Aims: To make three dimensional structure of native and mutated matrix protein (M) of Nipah Virus (NiV) and to establish conformational and functional comparison between the two. Study Design: All in-silico analysis were performed using various online and offline software. Place and Duration of Study: Department of Biotechnology and Bioinformatics, Padmashree Dr. D.Y.Patil Vidyapeeth, Navi Mumbai, February 2013. Methodology: The protein’s physicochemical properties were characterized and 3D model of both the normal and the mutated protein were created using Phyre2. Single point mutation of S147G is recorded which leads to altered structure formation. Both the models were evaluated and compared conformationally. Results: 4G1GB based structures were modeled by phyre2 and minimized energies recorded were-16760.041 kJ/mol for native and -16563.029 kJ/mol for the mutated protein. Structure validation proved that both the native and mutated structures were reliable. Formation of 3 H-bonds make mutated M structure slightly more stable than the native one. Conclusion: NiV, one of deadliest pathogen, needs to be checked immediately. More information gain is needed by performing wet lab analysis. This work might help understand the functional difference between native and mutated M protein and can be used as the potent drug target via applying rational drug designing approach.
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The incidence of emerging infectious diseases in humans has increased within the recent past or threatens to increase in the near future. Over 30 new infectious agents have been detected worldwide in the last three decades; 60 per cent of these are of zoonotic origin. Developing countries such as India suffer disproportionately from the burden of infectious diseases given the confluence of existing environmental, socio-economic, and demographic factors. In the recent past, India has seen outbreaks of eight organisms of emerging and re-emerging diseases in various parts of the country, six of these are of zoonotic origin. Prevention and control of emerging infectious diseases will increasingly require the application of sophisticated epidemiologic and molecular biologic technologies, changes in human behaviour, a national policy on early detection of and rapid response to emerging infections and a plan of action. WHO has made several recommendations for national response mechanisms. Many of these are in various stages of implementation in India. However, for a country of size and population of India, the emerging infections remain a real and present danger. A meaningful response must approach the problem at the systems level. A comprehensive national strategy on infectious diseases cutting across all relevant sectors with emphasis on strengthened surveillance, rapid response, partnership building and research to guide public policy is needed.
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During 2001-2011, multidisciplinary teams from the Institute of Epidemiology, Disease Control and Research (IEDCR) and International Centre for Diarrhoeal Disease Research, Bangladesh(icddr,b) identified sporadic cases and 11 outbreaks of Nipah encephalitis. Three outbreaks were detected through sentinel surveillance; others were identified through event-based surveillance. A total of 196 cases of Nipah encephalitis, in outbreaks, clusters and as isolated cases were detected from 20 districts of Bangladesh; out of them 150 (77%) cases died. Drinking raw date palm sap and contact with a case were identified as the major risk factors for acquiring the disease. Combination of surveillance systems and multidisciplinary outbreak investigations can be an effective strategy not only for detection of emerging infectious diseases but also for identification of novel characteristics and risk factors for these diseases in resource- poor settings.
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Nipah virus, a member of the genus Henipavirus, a new class of virus in the Paramyxoviridae family, has drawn attention as an emerging zoonotic virus in south east and south asian region. Case fatality rate of Nipah virus infection ranges from 40-70% although it has been as high as 100% in some outbreaks. Many of the outbreaks were attributed to pigs consuming fruits partially eaten by fruit bats, and transmission of infection to humans. In Bangladesh, 7 outbreaks of Nipah virus infection were identified during the period 2001–2007. In Bangladesh, Nipah virus infection was associated with contact with a sick cow, consumption of fresh date palm sap (potentially contaminated with pteropid bat saliva), and person-to-person transmission. In the most recent epidemic at least 15 people died due to Nipah virus infection in Hatibandha, Lalmonirhat district in a remote northern Bangladesh town in 2011 adding to the previous death toll of 113 in the country . Human infections range from asymptomatic infection to fatal encephalitis. Infected people initially develop influenzalike symptoms of fever, headaches, myalgia , vomiting and sore throat. This can be followed by dizziness, drowsiness, altered consciousness, and neurological signs that indicate acute encephalitis. Some people can also experience atypical pneumonia and severe respiratory problems. The virus is detected by ELISA, PCR, immunofluoroscent assay and isolation by cell culture. Treatment is mostly symptomatic and supportive as the effect of antiviral drugs is not satisfactory, and an effective vaccine is yet to be developed. So the very high case fatality addresses the need for adequate and strict control and preventive measures.
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The henipaviruses, represented by Nipah virus and Hendra virus, are emerging zoonotic viral pathogens responsible for repeated outbreaks associated with high morbidity and mortality in Australia, Southeast Asia, India and Bangladesh. These viruses enter host cells via a class I viral fusion mechanism mediated by their attachment and fusion envelope glycoproteins; efficient membrane fusion requires both these glycoproteins in conjunction with specific virus receptors present on susceptible host cells. The henipavirus attachment glycoprotein interacts with a cellular B class ephrin protein receptor triggering conformational alterations leading to the activation of the viral fusion (F) glycoprotein. The analysis of monoclonal antibody (mAb) reactivity with G has revealed measurable alterations in the antigenic structure of the glycoprotein following its binding interaction with receptor. These observations only appear to occur with full-length native G glycoprotein, which is a tetrameric oligomer, and not with soluble forms of G (sG), which are disulfide-linked dimers. Single amino acid mutations in a heptad repeat-like structure within the stalk domain of G can disrupt its association with F and subsequent membrane fusion promotion activity. Notably, these mutants of G also appear to confer a postreceptor bound conformation implicating the stalk domain as an important element in the G glycoprotein's structure and functional relationship with F. Together, these observations suggest fusion is dependent on a specific interaction between the F and G glycoproteins of the henipaviruses. Further, receptor binding induces measurable changes in the G glycoprotein that appear to be greatest in respect to the interactions between the pairs of dimers comprising its native tetrameric structure. These receptor-induced conformational changes may be associated with the G glycoprotein's promotion of the fusion activity of F.
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Objective:To investigate the epidemics of Nipah virus(NiV)in Yili, Xinjiang Uygur Autonomous Region. Methods:One step real time reverse transcriptase polymerase chain reaction(real-time RT-PCR)was performed to detect the NiV N in peripheral blood mononuclear cells from 120 equines. Gene sequence and amino acid sequence were analyzed for positive products.Results:The positive rate of NiV N in PBMCs from 120 equines was 0%(0/120).Conclusion:The investigation do not support that NiV infection exist in Yili, Xinjiang Uygur Autonomous Region.
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Objective To establish nucleic acid testing techniques for detecting Nipah virus (NiV) and Hendra virus (HeV), and to test the NiV and HeV in peripheral blood collected from domestic pigs, cows and goats in Chongqing. Methods Peripheral blood samples of 580 domestic pigs, 250 cows, 180 goats were collected from Chongqing since June 2007 to June 2008. The lymphocytes were separated by density gradient centrifugation and total RNA was extracted using Trizol method for detection of NiV and HeV with one-step real-time RT-PCR. Sequence identification and analysis were performed for positive PCR prod-ucts. Virus isolation and culture were adopted for positive samples, and epidemiologic reports were submit-ted. Results Nucleic acid detections searching for NiV and HeV were successfully performed in animal blood samples collected from Chongqing. "Takeoff points" were not found in fluorescence amplification curves of all samples. Curves kept the same slope, and assays were judged as negative. Conclusion Until now, Neither NiV or HeV infection has been found in domestic animals blood samples collected from Chongqing, which suggest a lower possibility of outbreaks of Nipah disease and Hendra disease in Chongqing in the near future.