Particle agglutination assay (PA) was developed and applied for detecting specific IgM antibodies to Japanese encephalitis virus in recent years. In this study, the sensitivity of PA and MAC-ELISA were 91.11 and 97.77%, respectively
Encephalitis, Japanese ; Encephalitis Viruses, Japanese ; diagnosis

Background: \r\n', u'Japanese encephalitis virus (JE) is the most prevalent in Asia, Pacific and in mainland northern Australia; and considered to be the leading cause to the acute encephalitis. The case- fatality and sequela rates in children stay rather high. There were some medical technologies for the JE diagnosis, of which is the application of the expensive MAC- ELISA and PEN TAX kits.\r\n', u'Objectives: \r\n', u'To evaluate and compare the light sensitivity of MAC- ELISA and PENTAX Kits in JE diagnosis.\r\n', u'Subjects and method: \r\n', u'48 pairs of sera samples obtained from the patients with clinically manifested Japanese encephalitis (JE) in 2003- 2005, in some northern provinces, they were applied and tested into MAC- ELISA Kits.\r\n', u'Results:\r\n', u'The sensitivity of MAC- ELISA and PENTAX kits detecting IgM against JE virus were 95.71%, and 98.57%, respectively. In addition to that, the sensitivity of these two kits used to detect JE IgM within the first 7 days of the disease was very high (around 92.31%-96.15%).\r\n', u'Conclusion: \r\n', u'The sensitivity of MAC- ELISA and PENTAX kits used to detect IgM against JE virus was very high. \r\n', u'
Encephalitis ; Japanese/ diagnosis

On 138 serum samples taken in the first 7 days of acute syndrome of brain, by MAC-ELISA technique 95 samples were determined as positive, 68.84% was positive. 43 samples taken in the first 7 days of the disease there was no detected antivirus IgM antibody but this could not concluded as negative, but may be a late antibody response or may be another cause of acute brain syndrome. For assuring the accuracy of the diagnosis, the samples taken in the first 7 days of the disease must be taken again after 7 day to verify the results.
Encephalitis, Japanese ; Diagnosis ; Enzyme-Linked Immunosorbent Assay

A retrospective study was carried out on 67 patients with Japanese Encephalitis at Dong Nai Pediatric Hospital from August 1999 to July 2001. Its showed a 40.8% of acute encephalitis patients admiited into hospital acquired Japanese encephalitisl Among them, 100% had IgM anti JE antibody(+), 100% were not vaccinated. Main clinical manifestations were fever 100%, coma 64.2%, vomit 71.6%, headache 55.2%, convulsion 46.2%. After treating: full recovery 61.2%, neuro-psychiatric sequella 11.9%, referral to higher level Pediatric Hospital 20.9%, death 6%. Because of the lack of specific therapeutic medicines, JE vaccine must be recommended to integrate to enlarged immunized program.
Encephalitis, Japanese ; Study Characteristics [Publication Type] ; Therapeutics ; diagnosis ; child ; hospitals

Fifteen pediatric cases of suspected Japanese encephalitis (JE) were reported in Beijing Children's Hospital during the late summer of 2013. The clinical manifestations in most cases included high fever, seizures, and abnormal magnetic resonance imaging findings. Twelve of 15 cases were laboratory-confirmed as JE cases by pathogen identification. Epidemiological investigations showed that five of the 12 laboratory-confirmed patients had an incomplete JE vaccination history. Follow-up investigations after discharge indicated that seven laboratory-confirmed JE patients without JE vaccinations had relatively poor prognoses, with an average Modified Rankin Scale (MRS) score of 2.6 when compared with the other five laboratory-confirmed, JE-vaccinated patients with an average MRS score of 0.5. The observation of pediatric JE cases among those with a history of JE vaccination warrants further attention.
Beijing ; epidemiology ; Child ; Child, Preschool ; Encephalitis Virus, Japanese ; physiology ; Encephalitis, Japanese ; diagnosis ; epidemiology ; virology ; Female ; Humans ; Japanese Encephalitis Vaccines ; administration & dosage ; Male ; Prognosis

Japanese encephalitis (JE) is a zoonosis that affects the nervous system of humans and other animals. The genotype of JE virus (JEV) has shifted recently from genotype 3 (G3) to genotype 1 (G1) in Asia, including Korea. Thus, a rapid differential assay is required to make an accurate diagnosis of JEV genotype. In this study, we designed common and differential primer sets for JEV G1 and G3 to detect the JEV envelope (E) gene. The specific primer sets for JEV G1 and G3 specifically amplified the target gene. The detection limits of the three primer sets were 10(1.0), 10(2.0), and 10(2.0) TCID₅₀/reaction, respectively. No cross-reactivity was detected with non-JEV reference viruses. The multiplex reverse transcription-polymerase chain reaction (RT-PCR) assay specifically differentiated JEV G1 from G3. Thus, a one-step multiplex RT-PCR assay was established to rapidly and differentially detect JEV. This assay will be useful for confirming JEV infections in animals and checking the JEV genotype in veterinary biological products.
Animals ; Asia ; Asian Continental Ancestry Group* ; Biological Products ; Diagnosis ; Encephalitis Virus, Japanese* ; Encephalitis, Japanese* ; Genotype ; Humans ; Korea ; Limit of Detection ; Nervous System

Objective: To investigate the distribution patterns of mosquitoes, midges and related arboviruses in Sichuan province. Methods: Blood-sucking insects were collected from houses and pens, using the ultraviolet lights. Mosquito samples were classified according to morphologic characteristics and then stored at liquid nitrogen. All samples were incubated with BHK-21 and C6/36 cells for virus isolation and then detected for their viral genes. Sequences of the virus were identified and analyzed by molecular biological software, such as BioEdit 7.0.5.3, MEGA 6.0. Results: In total, 17 019 mosquitoes from 3 genera and 4 species and 12 700 midges were collected from the southeast regions of Sichuan province in 2016 and 2017. Among them, 79.4% (13 519/17 019) belonged to Culex tritaeniorhynchus with 11.1% (1 897/17 019) as Armigeres subalbatus, 5.5% (930/17 019) were Anopheles sinensis and 4.0% (673/17 019) were Anopheles sinensis 3 virus strains that isolated from Culex tritaeniorhynchus were identified as typeⅠ Japanese encephalitis virus. Seven pools of mosquitoes isolated from Hejiang county were identified Japanese encephalitis virus gene positive through PCR amplification. With 4 pool midges were detected positive for Akabane virus through PCR gene amplification while midges samples didn't have virus isolates. Conclusions: Culex tritaeniorhynchus appeared the predominant species in the southeast regions of Sichuan. Japanese encephalitis virus transmitted by mosquitoes and Akabane virus by midges were prevalent in southeast Sichuan province.
Animals ; Arboviruses ; Culicidae ; Encephalitis Virus, Japanese/isolation & purification* ; Encephalitis, Japanese/diagnosis* ; Genes, Viral ; Nucleic Acid Amplification Techniques ; Phylogeny ; Polymerase Chain Reaction

BACKGROUNDTo compare the biological characteristics of West Nile virus (WNV) and Japanese encephalitis virus (JEV), including cells sensitivity, pathogenicity, viral morphology, as well as the results of immunological and molecular biological detection.

METHODSCytopathic effect (CPE) and pathogenicity were observed in C6/36 cells and in suckling mice inoculated intracerebrally with the WNV or JEV, respectively. The sliced tissue samples for electron microscopic examination were prepared for the morphologic observation of the viruses. Serum antibody to WNV or JEV was detected using indirect immunofluorescence assay (IFA), and the viral RNA was analyzed by RT-PCR method.

RESULTSWNV or JEV-caused CPE was characterized by cell fusion and cell shedding, respectively. There was no significant difference in the pathogenicity to suckling mice between WNV and JEV. The morphologic observation showed that the shape and size of the two virions were similar. WNV and JEV were found to have antigenic cross-reactivity. The viral RNA could be detected from both WNV and JEV samples with universal primer set, but only nucleoside fragments of corresponding virus could be amplified when specific primers were used.

CONCLUSIONCPE in C6/36 cell and detection of the viral RNA should be useful in discrimination of WNV and JEV, and simultaneously examining the titers of serum antibodies against WNV and JEV may be helpful to diagnosis of infection with these agents.

Animals ; Brain ; virology ; Cell Line ; Diagnosis, Differential ; Encephalitis Virus, Japanese ; immunology ; isolation & purification ; Encephalitis, Japanese ; diagnosis ; virology ; Flavivirus Infections ; diagnosis ; virology ; Immunoglobulin G ; blood ; Mice ; Mice, Inbred BALB C ; West Nile virus ; immunology ; isolation & purification

One step TaqMan reverse transcription polymerase chain reaction (RT-PCR) using TaqMan probe was developed for detection of Japanese encephalitis virus (JEV). Real-time RT-PCR was optimized to quantify JEV using the detection system (Rotor Gene 2000 detector) and dual-labeled fluorogenic probes. The gene specific labeled fluorogenic probe for the 3' non-translated region (3' NTR) was used to detect JEV. When the specificity of the assay using specific JEV primers was evaluated by testing three different JEV strains, other swine viruses and bovine viral diarrhea virus, no cross-reactions were detected with non-JE reference viruses. A single tube TaqMan assay was shown to be 10-fold more sensitive than the conventional two-step RT-PCR method. Detection limits of two step and real-time RT-PCR for JEV were 112 TCID50 /ml and 11.2 TCID50 /ml, respectively. Quantification of JEV was accomplished by a standard curve plotting cycle threshold values (Ct ) versus infectivity titer. Real-time RT-PCR assay using single tube method could be used as a sensitive diagnostic test, and supplied the results in real time for detection and quantification of JEV. We could detect JEV RNA genome in plasma samples of pigs inoculated with KV1899 strain at 2 days post inoculation, but couldn't in 41 fetus samples. This assay was sensitive, specific, rapid and quantitative for the detection of JEV from laboratory and field samples.
Animals ; DNA Primers/chemistry ; DNA Probes/chemistry ; Encephalitis Virus, Japanese/genetics/*isolation&purification ; Encephalitis, Japanese/diagnosis/*veterinary/virology ; RNA, Viral/analysis ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction/methods/*veterinary ; Sensitivity and Specificity ; Swine ; Swine Diseases/*diagnosis/virology ; *Taq Polymerase

Japanese encephalitis virus (JEV) is a member of Flaviviruses, transmitted by mosquitoes. The core of JEV is composed of the capsid (C) proteins. In order to produce the recombinant viral C protein and the antiserum specifically recognizing the JEV C protein, we have expressed and purified the JEV C protein as a Glutathion-S-Transferase (GST) fusion protein in E. coli. The JEV C protein-coding region was PCR-amplified using the infectious cDNA of a JEV Korean isolate, and the amplicons were cloned into the pGEX4T-1 E. coli expression vector. GST-C fusion proteins were purified using a glutathione sepharose column. Subsequently, the GST-C fusion proteins were used for immunization of rabbits, and the antisera were obtained from those immunized animals. Western blot analysis using the JEV-infected BHK21 cell lysates showed that these antisera specifically reacted with the JEV C proteins. This study will provide a useful reagent for the diagnosis and understanding of the viral morphogenesis in the JEV-infected cells.
Animals ; Asian Continental Ancestry Group* ; Blotting, Western ; Capsid Proteins* ; Capsid* ; Clone Cells ; Culicidae ; Diagnosis ; DNA, Complementary ; Encephalitis Virus, Japanese* ; Encephalitis, Japanese* ; Flavivirus ; Glutathione ; Humans ; Immune Sera ; Immunization ; Morphogenesis ; Rabbits ; Sepharose ; Staphylococcal Protein A