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

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

OBJECTIVETo explore Chinese medical syndrome distribution features of Japanese encephalitis (JE), and to analyze its correlation between syndromes and features of etiologies and pathogeneses.

METHODSRecruited were 277 patients with confirmative diagnosis of JE from Wuhan Medical Treatment Center, Children's Hospital Affiliated to Chongqing Medical University, Fifth People's Hospital of Guiyang City, Hangzhou Sixth People's Hospital, and Chengdu Hospital of Infectious Diseases between July to September 2012. Chinese medical syndrome distribution features were summarized from their general materials and detailed records of clinical data, including medical history, symptoms and signs, tongue fur, and pulse figures.The frequency of symptoms and signs was calculated according to mild, ordinary, severe, extreme severe degrees. The distribution of Chinese medical syndromes was summarized. And its correlation between syndromes and features of etiologies and pathogeneses were analyzed.

RESULTSAfter clustering analysis, Chinese medical syndromes of JE could be categorized as four groups: toxicity accumulation in Fei and Wei syndrome (TAFWS), brain collateral impaired by poison syndrome (BCIPS), depression of toxicity in the pericardium syndrome (DTPS), exhaustion of yin and yang syndrome (EYYS). BCIPS and DTPS were dominated, accounting for 74.0% (205 cases). The main causes covered evil of summer heat [accounting for 92.42% (256/277 cases)], heat [accounting for 87.73% (243/277 cases)], and toxin [accounting for 99.64% (276/277 cases)].

CONCLUSIONSThe four Chinese medical syndrome types of JE met Chinese medical clinical features of encephalitis. It is induced by infestation of dampness-heat, resulting in toxicity accumulation in Fei and Wei, brain collateral impaired by poison, depression of toxicity in the pericardium. Yin fluid and blood is exhausted as time goes by. Qi and yin are impaired to form intermingled deficiency and excess, and finally causing exhaustion of yin and yang.

Adolescent ; Child ; Child, Preschool ; Encephalitis, Japanese ; diagnosis ; pathology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Medicine, Chinese Traditional ; methods ; Yang Deficiency ; diagnosis ; Yin Deficiency ; diagnosis

Japanese encephalitis is one of the leading causes of acute encephalitis in Asia. But in Korea, the number of Japanese encephalitis cases has dropped considerably due to mass vaccination and vector control. Especially, there were no case reports under the age of 9 years during the last ten years. We will describe a case of a previously healthy 7-month old boy who presented with fever and seizure. The patient was diagnosed with Japanese encephalitis, based on the cerebrospinal fluid and serum antibody analyses for the Japanese encephalitis virus. Typical brain magnetic resonance image findings of Japanese encephalitis were observed. The patient received extensive conservative treatment including high dose intravenous corticosteroid treatment and immunoglobulin. In spite of severe hemodynamic instability, the patient survived, and he is currently in a vegetative state with respiratory assist by a home ventilator. Although the incidence of Japanese encephalitis dropped dramatically in Korea, pediatricians should always consider the diagnosis as one of the possibilities for patients with encephalitis, especially if the patient is not immunized for JEV. Since there is no specific treatment for JEV, timely and comprehensive conservative care is critical to reduce the mortality and morbidity.
Asia ; Asian Continental Ancestry Group* ; Brain ; Cerebrospinal Fluid ; Diagnosis ; Encephalitis ; Encephalitis Virus, Japanese ; Encephalitis, Japanese* ; Fever* ; Hemodynamics ; Humans ; Immunoglobulins ; Incidence ; Infant* ; Korea ; Male ; Mass Vaccination ; Mortality ; Persistent Vegetative State ; Seizures* ; Ventilators, Mechanical

OBJECTIVETo understand the epidemiological characteristics and the main clinical symptoms of viral encephalitis in Gansu.

METHODSA total of 322 viral encephalitis patients were recruited from province sentinel hospitals in Gansu province from 2009 to 2011, and their basic information were collected as well as their serum samples and cerebrospinal fluid samples. 296 out of the 322 cases were qualified for our study. Based on the patients' epidemiological characteristics and clinical features, we determined the detection of the virus types (at least one kind of virus detection was carried out for each case). ELISA was applied to test the IgM antibody of Japanese encephalitis (JE) virus (JEV), enterovirus (EV: including Coxsackie virus, echovirus, enterovirus 71), mumps virus and herpes simplex virus (HSV) in cerebrospinal fluid and serum specimen. The difference of positive detected rate between types of virus, among patients from different regions, time, or at different ages, as well as the different clinical symptoms between JE patients and other viral encephalitis patients, were analyzed and compared.

RESULTSThe positive detected rate of virus in the 296 patients was 27.03% (80/296); the positive rate of JEV, EV, mumps virus, HSV detected was separately 7.53% (22/292), 8.75% (23/263), 13.84% (22/159) and 15.09% (40/265), and the difference was statistically significant (χ(2) = 10.849, P < 0.05). 90.91% (20/22) of the JEV positive cases were distributed in Tianshui, Longnan and Pingliang, and 95.45% (21/22) patients were infected from July to September. All the 23 EV detected positive patients were infected from April to December, while the ages of patients ranged from 1 to 44 years old. Mumps virus, HSV testing positive cases had onset every month. Logistic regression analysis showed that the patients who had the symptoms as disturbance of consciousness (OR = 15.487, 95%CI: 2.266 - 105.852), somnolence (OR = 11.659, 95%CI: 1.783 - 76.242), convulsions (OR = 11.062, 95%CI: 1.687 - 72.530) were more likely to infect JEV.

CONCLUSIONHSV was the principal pathogen of viral encephalitis in Gansu. An obvious central tendency in the regional and time distribution was found in JEV infection; and the clinical symptoms of JE patients were more severe.

Adolescent ; Adult ; Antibodies, Viral ; blood ; cerebrospinal fluid ; Child ; Child, Preschool ; China ; epidemiology ; Encephalitis Virus, Japanese ; Encephalitis, Viral ; diagnosis ; epidemiology ; virology ; Enterovirus ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunoglobulin M ; blood ; cerebrospinal fluid ; Infant ; Male ; Mumps virus ; RNA, Viral ; blood ; cerebrospinal fluid ; Simplexvirus ; Young Adult

OBJECTIVETo develop a rapid, cost effective RT-PCR method for the mass scale diagnosis of such diseases at the viremia stage to find out the actual disease burden in that area.

METHODSFor this purpose, cases with the history of only short febrile illness were considered. Thus 157 samples with the history of dengue/chikungunya like illness and only 58 samples with a history of acute encephalitis syndrome (AES) were selected.

RESULTSOut of 157 samples, 42 and 74 were detected as dengue and chikungunya, respectively and out of 58 AES cases only 23 could be detected as Japanese encephalitis by this RT-PCR method.

CONCLUSIONSThis cost effective RT-PCR method can detect the total positive cases that remain undetected by ELISA method. Moreover, this method is capable to detect the viral RNA from patients' sera even after the appearance of IgM antibody at one fifth costs as compared with the other commercially available kits.

Antibodies, Viral ; blood ; Arbovirus Infections ; diagnosis ; virology ; Arboviruses ; genetics ; Chikungunya Fever ; diagnosis ; virology ; Dengue ; diagnosis ; virology ; Encephalitis Virus, Japanese ; genetics ; Encephalitis, Japanese ; diagnosis ; virology ; Fever ; diagnosis ; virology ; Humans ; Immunoglobulin M ; blood ; Mass Screening ; RNA, Viral ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; economics ; methods ; Sensitivity and Specificity ; Viremia ; diagnosis ; virology

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

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

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