Detection Of Japanese Encephalitis Virus Research Articles

Study on the temporal and spatial distribution of Culex mosquitoes in Hanoi, Vietnam

Arboviruses transmitted by mosquitoes, including Japanese encephalitis virus (JEV), present a substantial global health threat. JEV is transmitted by mosquitoes in the genus Culex, which are common in both urban and rural areas in Vietnam. In 2020, we conducted a 1-year survey of Culex mosquito abundance in urban, suburban, and peri-urban areas of Hanoi using CDC-light traps. Mosquitoes were identified to species and sorted into pools based on species, sex, and trap location. The mosquito pools were also investigated by RT-qPCR for detection of JEV. In total, 4829 mosquitoes were collected over a total of 455 trap-nights, across 13 months. Collected mosquitoes included Culex, Aedes, Anopheles, and Mansonia species. Culex mosquitoes, primarily Cx. quinquefasciatus, predominated, especially in peri-urban areas. Most Culex mosquitoes were caught in the early months of the year. The distribution and abundance of mosquitoes exhibited variations across urban, suburban, and peri-urban sites, emphasizing the influence of environmental factors such as degree of urbanization, temperature and humidity on Culex abundance. No JEV was detected in the mosquito pools. This study establishes baseline knowledge of Culex abundance and temporal variation, which is crucial for understanding the potential for JEV transmission in Hanoi.

Development of Colloidal Gold-Based Immunochromatographic Strips for Rapid Detection and Surveillance of Japanese Encephalitis Virus in Dogs across Shanghai, China.

Japanese encephalitis virus (JEV) causes acute encephalitis in humans and is of major public health concern in most Asian regions. Dogs are suitable sentinels for assessing the risk of JEV infection in humans. A neutralization test (NT) or an enzyme-linked immunosorbent assay (ELISA) is used for the serological detection of JEV in dogs; however, these tests have several limitations, and, thus, a more convenient and reliable alternative test is needed. In this study, a colloidal gold immunochromatographic strip (ICS), using a purified recombinant EDIII protein, was established for the serological survey of JEV infection in dogs. The results show that the ICSs could specifically detect JEV antibodies within 10 min without cross-reactions with antibodies against other canine viruses. The test strips could detect anti-JEV in serum with dilution up to 640 times, showing high sensitivity. The coincidence rate with the NT test was higher than 96.6%. Among 586 serum samples from dogs in Shanghai examined using the ICS test, 179 (29.98%) were found to be positive for JEV antibodies, and the high seropositivity of JEV in dogs in China was significantly correlated with the season and living environment. In summary, we developed an accurate and economical ICS for the rapid detection of anti-JEV in dog serum samples with great potential for the surveillance of JEV in dogs.

The detection of Japanese encephalitis virus in municipal wastewater during an acute disease outbreak.

To demonstrate the capability of wastewater-based surveillance (WBS) as a tool for detecting potential cases of Japanese Encephalitis Virus (JEV) infection in the community. In this study, we explore the potential of WBS to detect cases of JEV infection by leveraging from an established SARS-CoV-2 wastewater surveillance program. We describe the use of two reverse transcriptase quantitative polymerase chain reaction (RTqPCR) assays targeting JEV to screen archived samples from two wastewater treatment plants (WWTPs). JEV was detected in wastewater samples collected during a timeframe coinciding with a cluster of acute human encephalitis cases, alongside concurrent evidence of JEV detection in mosquito surveillance and the sentinel chicken programs within South Australia's Riverland and Murraylands regions. Current surveillance measures for JEV encounter multiple constraints, which may miss the early stages of JEV circulation or fail to capture the full extent of transmission. The detection of JEV in wastewater during a disease outbreak highlights the potential WBS has as a complementary layer to existing monitoring efforts forming part of the One Health approach required for optimal disease response and control.

High-throughput sequencing-based Detection of Japanese encephalitis virus and its effect on micro ribonucleic acid

it was to explore the mechanism of Japanese encephalitis virus (JEV) and micro ribonucleic acid (miRNA) under high-throughput sequencing. 20 experimental mice, with good growth status and no disease infection, were selected. The cells used in the experiment included mouse microglial cell line (BV2), mouse neuroblastoma cell line (NA), and mouse brain endothelial cell line (bEnd.3). JEV titration was performed with JEV-infected cells, ribonucleic acid (RNA) in the cells was extracted, and finally the miRNA high-throughput sequencing data was analyzed. Agarose gel electrophoresis showed that the 28S and 18S electrophoresis bands were bright. Among the miRNAs detected in mouse brain tissues, 2986 were down-regulated and 1251 were up-regulated. Among miRNAs detected in NA cells, 4238 the decreasing expression and 2356 were expressed increasingly. In reducing miRNA expression, 1 multiplicity of infection (MOI) of P3 strain infection was more significant than 0.1 MOI. 10 miRNAs with significantly decreasing expression were miR-466d-3p, miR-381–3p, miR-540–3p, miR-466a-3p, miR-467a-3p, miR-574–5p, miR-199a-5p, miR-467a-5p, miR-674–5p, and miR-376b-3p. These were all obviously down-regulated in JEV-infected BV2, NA, and bEnd.3 neurons. High-throughput sequencing of JEV-infected mouse brain tissues and mouse neuronal cells found that JEV infection led to down-regulation of overall miRNA expression in host cells.

A one-tube rapid visual CRISPR assay for the field detection of Japanese encephalitis virus

Early and rapid detection of Japanese encephalitis virus (JEV) is necessary for timely preventive and control measures. However, JEV RNA detection remains challenging due to the low level of viremia. In this study, a RApid VIsual CRISPR (RAVI-CRISPR) assay based on reverse transcription loop-mediated isothermal amplification (RT-LAMP) and CRISPR/Cas12a targeting was developed for easy detection of JEV in the field. We showed successful detection of 8.97 or more copies of the C gene sequence of JEV RNA within approximately 60 min. This assay also displayed no cross-reactivity with other porcine pathogens. We applied our one-tube RAVI-CRISPR assay to 18 brain tissue sample for JE diagnosis. The results from both fluorescence intensity measurements and directly naked-eye visualization were consistent with those from real-time PCR analysis. Taken together, our results showed that one-tube RAVI-CRISPR assay is robust, convenient, sensitive, specific, affordable, and potentially adaptable to on-site detection or surveillance of JEV in clinical and vector samples.

SERS based rapid and ultrasensitive detection of Japanese Encephalitis Virus

Japanese encephalitis (JE) is a mosquito-borne flavivirus infection named Japanese Encephalitis Virus (JEV), prevalent in Asia-pacific countries, requires an accurate and rapid diagnosis to contain the outbreak of the disease. In cases of low viral load in early-stage infections, this task becomes difficult. Therefore, we have developed a surface-enhanced Raman spectroscopy (SERS) based biosensor for rapid, sensitive, and early-stage detection of JE antigen. In this work, silver nanoparticles were deposited over a glass coverslip and used as a substrate for designing the sensing platform. Silver Nanoparticles have good metallic properties and plasmon activity. Therefore, it amplifies the Raman signals and provides a suitable surface for the SERS substrate. The developed platform has been used for the detection of the Japanese encephalitis virus (JEV). The fabricated sensor shows a linear response from 5 ng/mL to 80 ng/mL with a limit of detection (LoD) of ∼7.6 ng/mL. Therefore, this method could be a significant addition to the diagnostic modalities for early, sensitive, and specific diagnoses of JE antigen even at the nanogram level.

Detection of Japanese Encephalitis Virus & West Nile Virus in Suspected Population in Pakistan

Objectives: To investigate the presence of IgM and IgG in the sera of patients screened for Dengue and Chikungunya during Dengue outbreak 2015-2019 in Karachi, Pakistan. 
 Methods: Japanese encephalitis virus (JEV) and West Nile virus (WNV) are two neglected viral infections in Pakistan. A sero-survey was conducted to assess the exposure of the population to JEV and WNV in Pakistan. The patients screened negative for Dengue and Chikungunya but had acute undifferentiated febrile illness during Dengue outbreak 2015-19 in Karachi, Pakistan were included in the study. Total 115 human sera were subjected to ELISA for detection of IgM and IgG antibodies produced in response to West Nile virus and Japanese Encephalitis virus. 
 Results: Our results highlighted that the overall prevalence of Japanese Encephalitis and West Nile Virus in screened population was 76.52%. The sero-positivity for Japanese encephalitis virus was found in 50 samples whereas; the prevalence of West Nile virus (WNV) was detected in 38 samples. However, 21 samples with co-circulation were indicated in our results. 
 Conclusion: In conclusion, this is an alert for physicians to suspect WNV and JEV infection in patients with a dengue-like illness especially those patients who are dwelling or traveling to and from Pakistan. Therefore, there is a need of standardized diagnostic facilities and surveillance system at national level to avoid misdiagnosis and rule out the actual prevalence rate in Pakistan.

Signal enhancing gold nanorods (GNR) and antibody modified electrochemical nanosensor for ultrasensitive detection of Japanese Encephalitis Virus (JEV) secretory Non-Structural 1 (NS1) biomarker

Japanese Encephalitis Virus (JEV), a zoonotic infectious disease is a major concern in East-Asian and Western Pacific regions with no available therapy or fool-proof vaccines. Hence it is imperative to develop diagnostic assays for rapid mass screening for JEV biomarkers in human as well as pigs for better management of this recurring epidemic and guided therapy of infected patients. In this research work, we have exploited the advantages of gold nanorods (GNR) as signal enhancers which include ease of bioconjugation, on fabricated electrodes coupled with Non-Structural 1 (NS1) antibodies (Ab) of Japanese Encephalitis Virus (JEV) for ultrasensitive detection of JEV NS1 protein, which is found secreted in the circulatory system of infected individuals. Each fabrication, and conjugation step was thoroughly characterised and the electrode was optimised for various testing parameters. JEV NS1 antigen (Ag) recognition using the optimised electrode was carried out in buffer (detection limit- 0.36 fM) and spiked serum (detection limit- 0.53 fM) within the linear range of 1 µM to 1 fM. The fabricated electrode did not show any non-specific binding with Dengue Virus, West Nile Virus or Yellow Fever Virus NS1-Ag. The electrode also provided a stable rapid response within 30 s when reused up to 4 times, and could be stored at 4 °C up to 3 weeks without substantial variation in output. Hence, the developed electrode has future applications for miniaturisation into a rapid diagnostic device for clinical diagnosis of JEV.

Detection of Japanese encephalitis virus and its specific antibody in abnormal swine litters in Vietnam

Japanese encephalitis is considered an essential disease-causing swine reproductive failure in Asian countries; however, significant knowledge gaps remain about this problem in actual cases. This study examined 55 cases of reproductive failure in sows, including one abortion and 54 full-term farrowing litters, to find the Japanese encephalitis virus (JEV) and specific antibodies against JEV. Haemagglutination test was used to detect specific antibodies against JEV from 63 samples of fetal thoracic fluids and newborn un-suckling piglet sera (54 thoracic fluid samples of dead fetuses, two thoracic fluid samples of abortion fetuses, seven sera samples of weakly newborn un-suckling piglets), viral isolation and RT- PCR technique was attempted from 60 brains of fetuses and newborn un-suckling piglets, histopathology sections of fetal brains which were positive with JEV confirmed by HI test and RT-PCR were also made. The results showed that 17.46% (11/63) of thoracic fluids and sera were positive for JEV. No JEV isolation was found from 60 brain samples of dead fetuses and piglets, but RNA of JEV were detected from 5 of them (8.33%). Our results suggest that JEV should be considered the important cause of swine reproductive failure in Viet Nam.

Rapid Differential Detection of Japanese Encephalitis Virus and Getah Virus in Pigs or Mosquitos by a Duplex TaqMan Real-Time RT-PCR Assay.

Both JEV (Japanese encephalitis virus) and GETV (Getah virus) pose huge threats to the safety of animals and public health. Pigs and mosquitoes play a primary role in JEV and GETV transmission. However, there is no way to quickly distinguish between JEV and GETV. In this study, we established a one-step duplex TaqMan RT-qPCR for rapid identification and detection of JEV and GETV. Primers and probes located in the NS1 gene of JEV and the E2 gene of GETV that could specifically distinguish JEV from GETV were selected for duplex TaqMan RT-qPCR. In duplex real-time RT-qPCR detection, the correlation coefficients (R2) of the two viruses were higher than 0.999. The RT-qPCR assay demonstrated high sensitivity, extreme specificity, and excellent repeatability. Detection of JEV and GETV in field mosquito and pig samples was 100 times and 10 times more sensitive than using traditional PCR, respectively. In addition, the new test took less time and could be completed in under an hour. Clinical sample testing revealed the prevalence of JEV and GETV in mosquitoes and pig herds in China. This complete duplex TaqMan RT-qPCR assay provided a fast, efficient, specific, and sensitive tool for the detection and differentiation of JEV and GETV.

First report on the detection of Japanese encephalitis virus in fruit bats from India.

Japanese encephalitis (JE) is a mosquito borne viral zoonotic disease and JE virus (JEV) is responsible for causing several children deaths every year in India. Since 1978, cases of JE have been reported from Gorakhpur district of Uttar Pradesh state annually. The knowledge on the role played by wildlife reservoirs in the sylvatic transmission and maintenance of JE virus remains limited. Bats are reservoir hosts for several emerging and re-emerging viral pathogens but their role in zoonotic cycle of JEV has not been elucidated yet. In Gorakhpur district of Uttar Pradesh, 52 fruit bats were found dead on 26 May 2020. The post-mortem report of the bat samples conducted at the Indian Veterinary Research Institute stated that the bats died due to brain hemorrhage, caused by excessive heat. The brain tissue samples of the bats were subjected to investigation using molecular techniques to determine the presence of JEV. The present work reports for the first time the detection of JEV in brain samples of bats from India. The viral load ranging from 8 to 18 copies/reaction was detected in brain samples by TaqMan real Time RT-PCR. The low viral load might be the reason for the absence of apparent clinical signs in bats and suggests the probable role of fruit bats in maintaining the JEV in nature.

Electroactive reduced graphene oxide for highly sensitive detection of secretory non-structural 1 protein: A potential diagnostic biomarker for Japanese encephalitis virus

Fluorine Doped Tin Oxide (FTO) electrode was fabricated with reduced Graphene Oxide (rGO) for sensitive detection of Japanese encephalitis virus (JEV) non-structural 1 (NS1) protein. Beforehand, in-silico 3D structure, stability, and docking of recombinant JEV NS1 antigen (NS1–Ag) and antibody (Ab) was evaluated. The recombinant NS1 Ag of 42 kDa was produced in-house by successful cloning into pET-28a(+) plasmid and further expressed using BL21 Escherichia coli (E. coli) cells. The NS1 Ag was used to raise polyclonal antibodies (Ab) and both were characterized via Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE), Western Blot, Matrix-Assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF), and Enzyme-Linked Immunosorbent Assay (ELISA). Further characterisation of all binding events such as rGO synthesis, and its conjugation with NS1 Ab, and NS1 Ag were confirmed through Fourier-Transform Infrared Spectroscopy (FTIR), Raman Spectroscopy, Energy Dispersive X-Ray Analysis (EDX), Scanning Electron Microscopy (SEM), Cyclic Voltammetry (CV) and Differential Pulse Voltammetry (DPV). The fabricated FTO electrode was optimised for various parameters such as pH, response time, temperature, concentration, and scan rate. The detection of JEV NS1 Ag was performed in buffer (LOD- 0.92 fM) as well in spiked serum (LOD- 1.3 fM) samples. The JEV NS1 Ab showed negligible cross-reactivity with other flaviviral NS1 Ag, provided a rapid response within 5 s, and remained stable up to 4 weeks. Furthermore, the fabricated immunosensor may be a potential candidate for further miniaturisation for accurate and early diagnosis of JEV in clinical samples.

The detection of Japanese encephalitis virus in Megachiropteran bats in West Kalimantan, Indonesia: A potential enzootic transmission pattern in the absence of pig holdings

The West Kalimantan province in Borneo island, Indonesia belongs to endemic area of Japanese encephalitis (JE) that accounts for approximately 30% of total cases yearly. As the presence of pig holdings is uncommon in West Kalimantan, another reservoir host might have played a role in the local transmission of JE virus in this area. Current study aimed to identify the potential role of bats in the local transmission of JE by performing molecular detection of JE virus in bats and mosquitoes using RT-PCR. Sample collection was performed in 3 districts in West Kalimantan, covering 3 different ecosystems: forest, coastal, and residential areas. Bat collection was performed using mist net and harp net, while mosquito collection was carried out using animal-baited trap and human landing collection. A total of 373 blood samples from bats were tested for JE virus, among which 21 samples (5.6%) showed positive results, mainly from Cynopterus brachyotis (lesser short-nosed fruit bat) found in residential areas. Out of 53 mosquito pools, 3 JE-positive pools of Culex tritaeniorhynchus and Cx. vishnui were collected at the same location as JE-positive bats. Current study showed the first evidence of JE virus detection in several species of Megachiropteran bats in Indonesia, demonstrated the potential role of frugivorous bats in local transmission of JE in West Kalimantan. More aggressive measures are required in JE risk mitigation, particularly in initiating JE vaccination campaign and in avoiding disruption of bats’ natural habitats through changes in land-use.

Rapid and simultaneous detection of Japanese encephalitis virus by real-time nucleic acid sequence-based amplification

Japaneses encephalitis (JE) is most common zoonoses caused by Japanese encephalitis virus (JEV) with a high mortality and disability rate. To take timely preventive and control measures, early and rapid detection of JE RNA is necessary. But due to characteristic brief and low viraemia, JE RNA detection remains challenging. In this study, a real-time nucleic acid sequence-based amplification (RT-NASBA) was developed for rapid and simultaneous detection of JEV. Four pairs of primer were designed using a multiple genome alignment of all JEV strains from GenBank. NASBA assay established and optimal reaction conditions were confirmed by using primers and probe on ns1 gene of JEV. The specificity and sensitivity of the assay were compared with RT-PCR by using serial RNA and virus cultivation dilutions. The results showed that JEV RT-NASBA assay was established, and robust signals could be observed in 10 min with high specificity. The limit of dectetion of RT-NASBA was 6 copies per reaction. The assay was thus 100 to 1, 000 times more sensitive than RT-PCR. The cross-reaction was performed with other porcine pathogens, and negative amplification results indicated the high specificity of this method. The novel JEV RT-NASBA assay could be used as an efficient molecular biology tool to diagnose JEV, which would facilitate the surveillance of reproductive failure disease in swine and would be beneficial for public health security.

Detection of recent infection of Japanese encephalitis virus in swine population using IgM ELISA: A suitable sentinel to predict infection in humans

Japanese encephalitis (JE) is a mosquito-borne flaviviral zoonotic disease and is one of the major causes of encephalitis in children. Swine, being an amplifier host of Japanese encephalitis virus (JEV), play an important role in its epidemiology. Therefore, early detection of either JEV or antibodies against JEV in swine is a feasible alternative for initiating necessary measures to prevent the spread of infection to humans. Since IgM antibodies appear early in swine sera, recombinant NS1 protein based indirect IgM ELISA was developed in the present study with the objective to know the recent infection of swine population with JEV. The relative diagnostic sensitivity and specificity of the developed ELISA was 95.34% and 98.6%, respectively. The developed ELISA was found to have excellent reproducibility on inter-laboratory and inter-institutional validation studies. A total of 3,027 field swine sera samples were screened using the developed ELISA and 488 samples were found positive for IgM against JEV with an overall sero-positivity of 16.12% in swine population of India. The highest sero-positivity was observed in swine population of Eastern zone of India which coincided with the maximum number of human JE cases reported from this zone during the same period. Further, antibody kinetics study revealed that the IgM antibodies against NS1 protein of JEV started appearing in swine sera at day 5 and disappeared completely by day 40. The IgG antibodies started appearing at day 7, and remained for more than 365 days indicating the suitability of IgM ELISA to know the recent infection of JEV. The developed IgM ELISA can be readily incorporated into surveillance programs for detection of JEV activity in swine population so that outbreaks in humans can be prevented by taking suitable preventive measures.

Detection of Japanese encephalitis virus in mosquitoes from Xinjiang during next-generation sequencing arboviral surveillance.

A total of 548 mosquitoes were collected from different animal farms located near to highly populated cities in Xinjiang and were subjected to metagenomic next-generation sequencing (mNGS). The mNGS data demonstrated that 18,842 (XJ1 strain) and 1,077 (XJ2 strain) of Japanese encephalitis virus (JEV)-related reads were detected in XJ1 and XJ2 mosquito samples collected from Wushi and Wensu counties of Aksu area, which accounted for 0.032% and 0.006% of the total clean reads generated from XJ1 and XJ2 samples, respectively. The Bayesian molecular phylogenetic analysis suggested that XJ1 and XJ2 strains belonged to JEV genotype III and were clustered with JEV strains isolated in China. Notably, Bayesian molecular time line phylogeny revealed that XJ1 strain shared its MRCA with JEV GSS strain about 67 YA, suggesting that XJ1 strain likely originated from linages closely related to GSS strain and spread to Xinjiang later. Overall, these findings suggest that Xinjiang was probably not free from JEV, and thus, a further surveillance of JEV is required in Xinjiang.

Fluorescent labeling based acoustofluidic screening of Japanese encephalitis virus

The Japanese encephalitis virus (JEV) as one of the main causes of encephalitis in Asia has resulted in tens of thousands of deaths. Early and efficient diagnosis of the virus is essential to prevent the spread of JEV. Here, we present a fluorescent labeling based acoustofluidic method for JEV screening. The carboxy-functionlised polystyrene (PS) microspheres were conjugated with monoclonal anti-JEV antibody (mAb) and then captured JEV in a complex biological samples. The composites of PS-mAb-JEV were separated in the microfluidic device via travelling surface acoustic (TSAW) force. The separated composites were labelled with FITC-conjugate anti-JEV antibody 2H4 and then were detected by the fluorescent signal. Experimental results proved the highly efficient separation and specific detection of JEV in the biological sample. Also, the acoustic-wave based separation method can separate efficiently almost all types of particles and has little effect on the subsequent genetic testing and fluorescence detection of JEV. Meantime, the fluorescence labeling method for JEV detection doesn’t need the laborious pretreatment and capable of offering a rapid and reliable detection approach. This is the first time to detect virus based on acoustofluidic driving and fluorescent labeling, which is a promising method for screening virus in the future biomedical applications.

Detection of Japanese Encephalitis Virus in Culex Mosquitoes in Singapore.

.Mosquito-borne flaviviruses are emerging pathogens of an increasing global public health concern because of their rapid increase in geographical range and the impact of climate change. Japanese encephalitis virus (JEV) and West Nile virus (WNV) are of concern because of the risk of reemergence and introduction by migratory birds. In Singapore, human WNV infection has never been reported and human JEV infection is rare. Four sentinel vector surveillance sites were established in Singapore to understand the potential risk posed by these viruses. Surveillance was carried out from August 2011 to December 2012 at Pulau Ubin, from March 2011 to March 2013 at an Avian Sanctuary (AS), from December 2010 from October 2012 at Murai Farmway, and from December 2010 to December 2013 at a nature reserve. The present study revealed active JEV transmission in Singapore through the detection of JEV genotype II in Culex tritaeniorhynchus collected from an Avian Sanctuary. Culex flavivirus (CxFV), similar to the Quang Binh virus isolated from Cx. tritaeniorhynchus in Vietnam and CxFV-LSFlaviV-A20-09 virus isolated in China, was also detected in Culex spp. (vishnui subgroup). No WNV was detected. This study demonstrates the important role that surveillance plays in public health and strongly suggests the circulation of JEV among wildlife in Singapore, despite the absence of reported human cases. A One Health approach involving surveillance, the collaboration between public health and wildlife managers, and control of mosquito populations remains the key measures in risk mitigation of JEV transmission in the enzootic cycle between birds and mosquitoes.

Molecular epidemiology of Japanese encephalitis virus in pig population of Odisha, Assam and Manipur states of India

Japanese encephalitis virus (JEV) comes under the family Flaviviridae and genus flavivirus. Pigs act as reservoir and amplifying intermediate host for JEV. The current investigation was conducted to understand the prevalence of JEV infection in pigs in three different geographical sites in India (Odisha, Assam and Manipur). Total 857 serum samples were tested by ELISA and RT-PCR, while only RT-PCR was performed in case of 275 tonsils tissues for detection of JEV. It was observed that JEV prevalence was highest in Manipur (positive 39, 25.5% in serum and 10% in tonsil) but lower in Assam (positive 15, 3.8% in serum and 0% in tonsils) and Odisha (positive 7, 1.5% in serum and 3.7% in tonsils). Genotype III (GIII) of JEV was the dominant genotype. Further, analysis of E gene revealed sporadic mutations of S83G, H76P, E78Q, C55S, and S64W along with two consistent mutations V46S and V51I in GIII. Whereas, a single mutation S118N was observed in the GI strain. In conclusion, the high JE virus infection rate of pig in the current locations suggests the need for continuous surveillance of this virus in pigs which will ultimately help to adopt an effective control strategy to prevent the spread of JE infection to human.

Fast and sensitive detection of Japanese encephalitis virus based on a magnetic molecular imprinted polymer–resonance light scattering sensor

A magnetic surface molecularly imprinted-resonance light scattering sensor was developed for rapid and highly sensitive detection of Japanese encephalitis virus (JEV). To prepare the surface imprinted polymer, Fe3O4 microspheres were selected as imprinting substrates which coated by silicon. Aminopropyl-triethoxysilane (APTES) as functional monomers for fixing template molecules JEV through a polymerization process of tetraethyl-orthosilicate (TEOS). The target virus JEV could be captured by the imprinted particles fastly and selectively, resulting in an increase of the RLS intensity. The results of RLS analysis proved that the obtained imprinted nanoparticles exhibited excellent specific recognition ability and high selectivity for the template virus (JEV). Furthermore, the response time of the sensor is within 20 min, which is much shorter than the previous works. The sensor with convenient separation and the limit of detection was 1.3 pM. These experimental results show that the proposed strategy is expected to achieve rapid and sensitive detection of JEV in practical applications.