Animals ; Bunyaviridae ; Bunyaviridae Infections ; Humans

Hantaviruses are negative-strand RNA viruses that contain three segmented genome and belong to the family Bunyaviridae. Due to such an unique structure of segmented RNA genome, these viruses have a possibility to produce reassortant that have genomic sets mixed with different segments originated from both parental virus during the genetic interaction. To investigate whether this phenomenon occurs actually, Hantaan virus (HTN) could be coinfected with Seoul virus (SEO) to Vero-E6 cell. And also, HTN could be coinfected with Prospect hill virus (PH) to investigate the rate of genetic reassortment according to the genetic distance among the HTN, SEO and PH viruses. As a available method to differentiate the reassortant, we used the multiplex RT-PCR that detect and differentiate the serotype of hantaviruses in mixed infection. Each progeny clone could be screened by multiplex RT-PCR. So, we have constructed the multiplex RT-PCR system that separated amplifications for each segment of Hantaviruses. Our multiplex RT-PCR system provide sensitive, specific and simplified tools for the rapid differentiation of hantaviruses serotypes and also the diagnosis of Hantavirus infections.
Bunyaviridae ; Clone Cells ; Coinfection ; Diagnosis ; Genome ; Hantaan virus ; Hantavirus Infections ; Hantavirus* ; Humans ; Hydrogen-Ion Concentration ; Parents ; RNA ; RNA Viruses ; Seoul virus ; Serotyping*

Hemorrhagic fever with renal syndrome (HFRS) is an acute viral disease with fever, hemorrhage and renal failure caused by hantavirus infection. Hantavirus induces HFRS or hantavirus pulmonary syndrome (HPS). HPS progression to a life-threatening pulmonary disease is found primarily in the USA and very rarely in South Korea. Here, we report a case of HFRS and coexisting HPS.
Fever ; Hantavirus ; Hantavirus Infections ; Hantavirus Pulmonary Syndrome ; Hemorrhage ; Hemorrhagic Fever with Renal Syndrome ; Lung Diseases ; Renal Insufficiency ; Republic of Korea ; Virus Diseases

Hantaviruses belong to the genus Hantavirus and Hantaan, Seoul, Puumala, Belgrade and Sin Nombre viruses are the etiolgic agents of two serious hantaviral diseases of humans. The rodent hosts and the specific etiologic agents of haemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS) are known and many reported cases occurred in Eurasia and Americas. Wild rodents trapped in 13 different areas of Korea from 1994 to 1998 were investigated against hantavirus infection. A total of 718 wild rodents and 10 species were trapped and found 630 (87.7%) of them were Apodemus agrarius. Indirect immunofluorescent antibody technique (IFAT) was performed for hantaviruses infections using different hantavirus antigens. Hantavirus antibodies were found in 68 (10.8%) out of 630 A. agrarius, 8 (42.1%) of 19 Rattus norvegicus. Among 68 lungs and other tissues of antibody positive A. agrarius, 5 (7.4%) were antigen positive. IFA titers of 5 positive A. agrarius sera showed higher titers against Puumala or Sin Nombre viruses than Hantaan virus. These results suggest that there may be are possibilities of existence of a noble hantavirus in Korean wild rodents.
Americas ; Animals ; Antibodies ; Fever ; Hantaan virus ; Hantavirus Infections* ; Hantavirus Pulmonary Syndrome ; Hantavirus* ; Hemorrhagic Fever with Renal Syndrome ; Humans ; Korea* ; Lung ; Murinae ; Rats ; Rodentia* ; Seoul ; Sin Nombre virus

OBJECTIVETo sequence the whole genome and to analyze the molecular and evolutionary of Severe fever with thrombocytopenia syndrome bunyavirus (SFTSV) isolated from Zhejiang province.

METHODSViral RNA was extracted from the specimens and detected by quantitative real-time RT-PCR. SFTSV strain was isolated. A total of 17 overlapping fragments covering the whole genome were amplified by RT-PCR. And the entire genomes were formed by sequencing and assembly the fragments. The SFTSV sequence of Zhejiang strain was compared with the sequences of SFTSV that have been published to generate the phylogenetic tree. And the SFTSV sequence of Zhejiang strain was compared with the sequences of strains of the genus phlebovirus in the Bunyaviridae family and analysis of homology.

RESULTSSFTSV strain was isolated from SFTSV infection positive serum successfully. The genomic fragments were amplified by RT-PCR. A total of 3 cDNA sections were formed by sequencing and assembly the fragments. The S segment contained 1 745 nucleotides. The M fragment contained 3 378 nucleotides, and the L segment contained 6 368 nucleotides. Molecular phylogenetic analysis result showed SFTSV Zhejiang strain had the highest similarity with Japan/SPL004A/2013 strain. The similarity of the S segment was 98%. The similarity of the M fragment was 97%. And it was 98% that of the L fragment. Meanwhile, the comparison results also confirmed the Zhejiang strain belonged to the genus phlebovirus.

CONCLUSIONSFTSV Zhejiang strain of isolated from SFTSV infection positive serum successfully. And the genome sequencing was complete molecular evolution analysis shows SFTSV Zhejiang strain has the maximum similarity with SFTSV Japan strain.

Base Sequence ; Bunyaviridae ; Bunyaviridae Infections ; Evolution, Molecular ; Genome ; Phlebovirus ; Phylogeny ; RNA, Viral ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo understand the infection status and epidemic rule of new bunia virus in the livestock and poultry which are closely related with humans such as sheep, cattle, dogs, pigs and chicken in the hilly area of Jiaodong peninsula in Shandong province.

METHODSPenglai and Laizhou in the hilly area of Jiaodong peninsula in Shandong province where severe fever with thrombocytopenia syndrome cases occurred in 2010 were selected as experimental sites. During April to November in 2011, serum specimens of the sheep, cattle, dogs, pigs and chicken with ticks in endemic area were randomly collected by random number table.5 ml venous blood was collected in each livestocks or poultries and there were total 3576 samples.New bunia virus antibody in different species of livestocks or poultries serum was continuously detected using double antigen sandwich enzyme-linked immunosorbent assay, and the infection rates of new bunia virus between different species of livestocks or poultries and between Penglai and Laizhou were analyzed using chi-square test.

RESULTSTest results in 3576 samples of livestocks or poultries serum specimen showed that the infection rate was as high as 63% (636/1013) in sheep, 53% (444/841)in cattle, 46% (242/530) in chicken, 29% (104/362)in the dogs, and 1% (12/830) in pigs. There were significant differences of new bunia virus infection among different species (χ(2) = 815.26, P < 0.05).In Penglai, the infection rate was as high as 71% (400/563) in sheep, 57% (232/409)in cattle, 35% (93/266) in chicken, 44% (796/1819)in total, while in Laizhou, the infection rate was 53% (236/450)in sheep, 49% (212/432)in cattle, 56% (149/264)in chicken, 36% (642/1757)in total, their difference was statistically significant(χ(2) values were 37.04, 4.93, 24.63, 19.38, all P values were < 0.05).Infection rates of dogs and pigs showed no obvious fluctuation.However, there were two peaks of infection in sheep in summer and autumn, the infection rate was as high as 62% (68/110) in June and 86% (204/236) in November;There were two peaks of infection in cattle in spring and autumn, the infection rate was as high as 56% (53/94) in April and 73% (116/159) in November; there was only one peak of infection in chicken, the infection rate was as high as 65% (55/85) in September.

CONCLUSIONThe infection rate is higher in sheep, cattle, chickens and dogs in the hilly area of Jiaodong peninsula. The peak season is spring, summer and autumn.

Animals ; Bunyaviridae ; isolation & purification ; Bunyaviridae Infections ; epidemiology ; veterinary ; Cattle ; Chickens ; China ; epidemiology ; Dogs ; Livestock ; virology ; Poultry ; virology ; Sheep

Bunyaviridae Infections ; pathology ; Fever ; virology ; Humans ; Leukopenia ; virology ; Orthobunyavirus ; classification ; pathogenicity ; Thrombocytopenia ; virology

OBJECTIVETo analyze and summarize the clinical characteristics, experience of diagnosis and treatment of cases infected by new bunyavirus, which occurred in Henan province in 2010.

METHODSThe clinical characteristics and effect of diagnosis and treatment of 5 cases were analyzed using descriptive epidemiological method. Blood specimens were detected by RT-PCR and pathogen separation.

RESULTSPCR testing was positive for all 5 cases. New bunyavirus were isolated from 2 cases. In 5 cases, fever (5/5), the whole body aches (5/5), fatigue (5/5), anorexia (5/5), nausea (5/5), the chills (4/5), cough (4/5), expectoration (4/5), vomiting (3/5), conjunctival hyperemia (3/5); Leukocyte reduction (5/5), thrombocytopenia (5/5), elevated alanine aminotransferase (4/5), elevated aspartate aminotransferase (4/5), elevated lactate dehydrogenase (5/5), creatine kinase elevations (4/5), urinary protein (3/5). By symptomatic and supportive treatment and prophylactic antibiotics, the first case died and the other 4 cases were cured. The average course of disease was 15.4 days.

CONCLUSIONCases infected by new bunyavirus have complicated clinical feature and multiple organ damage. If symptomatic treatment is in time, prognosis will be good.

Adult ; Bunyaviridae Infections ; diagnosis ; therapy ; virology ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Orthobunyavirus ; Prognosis ; Young Adult

OBJECTIVETo learn the prevalence of infection of human and animals severe fever with thrombocytopenia syndrome bunyavirus (SFTSV) in Yantai, Shandong province, and to analyze the pathogenic features of SFTSV as well as its relationship between human and animal hosts.

METHODSFrom April to November in 2011, 3 576 serum samples were collected from domesticated animals, including sheep, cattle, pigs, dogs, chickens, in Laizhou and Penglai areas where fever with thrombocytopenia syndrome frequently occurred among local residents. Total SFTSV antibodies and virus-specific nucleic acids of the serum were tested by ELISA and Real time RT-PCR, respectively. SFTSV infection on each animal was observed in different months. 2 590 human serum samples were also collected in Laizhou and Penglai areas, with IgG antibodies tested by ELISA. Virus was isolated with Vero cells from the serum which SFTSV viral nucleic acids were positive. S fragments were amplified by RT-PCR and sequenced, with homology analysis conducted on these sequences.

RESULTSThe overall positive rate of serum samples from animals on the total SFTSV antibodies was 40.24% (1 439/3 576) while the positive rate for specific nucleic acids was 4.56% (163/3 576). The positive rates for SFTSV antibodies were 62.78%, 52.97%, 45.56%, 28.73%, 1.45% and the positive rates for specific nucleic acids were 5.72%, 4.63%, 3.02%, 5.25% and 3.73%, in sheep, cattle, chickens, dogs, pigs, respectively. The antigens/antibodies for SFTSV in animals changed seasonally. The overall positive rate for SFTSV IgG antibody from 2 590 human samples was 5.41%. Thirteen virus strains were isolated from these serum samples (10 strains from human and 3 strains from animals). The nucleotide homology of 13S fragments' sequences ranged from 95.23% to 100.00% and the nucleotide homology with the isolates from other provinces were between 94.72% and 99.13%. The homology was considered to be high.

CONCLUSIONHigh prevalence of SFTSV infections occurred both in human and domestic animals in Yantai city. The nucleotide sequences of SFTSV were highly homologous among human and domestic animals. The findings suggested that domesticated animals might serve as SFTSV proliferation and the hosts for transmission thus should be attached great importance.

Animals ; Antibodies, Viral ; blood ; Bunyaviridae Infections ; epidemiology ; China ; epidemiology ; Humans ; Prevalence ; RNA, Viral ; blood ; Sequence Analysis, RNA

Severe fever with thrombocytopenia syndrome bunyavirus is a newly emerging virus in China, enveloped with a tripartite, single-stranded RNA genome of negative polarity. The regulatory elements for viral transcription and replication, as well as encapsidation and packaging signals, are thought to be located within these noncoding regions (NCRs). The terminal nucleotides are genus specific and highly conserved. The function of the remaining nucleotides of the NCRs is still not well understood. In this study, we developed the plasmid-driven RNA polymerase I minireplicon system for SFTSV firstly, using reporter genes GFP and luciferase. The function of the noncoding regions of the three Bunyaviridae RNA segments (L, M, S) in transcription was analyzed. Reporter genes are successfully expressed in SFTSV minireplicon system. Our results suggest that the NCRs of SFTSV from all three segments contain the necessary signals to initiate transcription. Quantitative detection of the luciferase expression level shows that promoter activity in the three segments is different.
Bunyaviridae Infections ; virology ; Cloning, Molecular ; Genome, Viral ; Humans ; Phlebovirus ; genetics ; physiology ; Replicon ; Viral Proteins ; genetics ; metabolism ; Virus Replication