Evaluation of a radioimmunoassay for type III procollagen peptide in the dog

Type III procollagen peptide (P-3-P) is a serum marker for hepatic fibrosis in humans. The utility of a commercially available radioimmunoassay for P-3-P was evaluated in the dog. The specificity of the assay was assessed by polyacrylamide gel electrophoresis (PAGE) of canine serum and purified bovine P-3-P, followed by Western immunoblotting with rabbit aniti-P-3-P serum. The sensitivity was assessed by performing the radioimmunoassay on dilutions of sera from 22 dogs. Polyacrylamide gel electrophoresis of purified bovine P-3-P and sera from two dogs suspected of having elevated P-3-P concentrations revealed no homologous bands of staining. Western immunoblotting showed marked cross-reactivity of the high antisera concentrations with several components of the serum proteins, but none corresponding to the purified P-3-P. All tested sera from dogs had minimal competitive binding with radiolabeled P-3-P in the radioimmunoassay. Dilution curves of dog sera did not parallel either the standard curve or the dilution curve of a known test human serum. There were no statistically different P-3-P concentrations in any of the groups of dogs studied. It was concluded that currently available radioimmunoassay kits for the measurement of P-3-P in the human are not applicable in the dog. Seemingly, the structure or metabolism of canine P-3-P may vary significantly from that of the bovine or human, limiting the sensitivity and specificity of this assay in the dog.     

草莓白化相关病毒中国分离物全基因组分析

草莓白化相关病毒(strawberrypallidosis-associatedvirus,SPa V)属于长线形病毒科(Closteroviridae)毛形病毒属(Crinivirus),可引起草莓病害,2017年在中国首次报道。采用高通量测序、RACE和RT-PCR技术获得了SPa V中国分离物(FJ)的基因组全长。该病毒含有两条正单链基因组RNA1和RNA2。RNA1全长8 048 nt,5′和3′非编码区序列分别为264和197 nt,含有3个开放阅读框(ORF),分别编码ORF 1a/1b融合蛋白和p9蛋白。RNA2全长7 977 nt,5′和3′非编码区序列分别为248和186 nt,含有8个开放阅读框(ORF),分别编码HSP70h、CPh、CP、CPm、p7、p6、p9和p28等8个蛋白。RNA1和RNA2与美国M1分离物分别具有98.5%和99.0%的核苷酸一致性;系统发育分析结果表明,SPa V中国分离物(FJ)单独处在一个分支。对SPa V来源的小RNA的分析表明,来源于SPa V的小RAN长度以21和22 nt为主。     

Faecal virome of red foxes from peri-urban areas

Red foxes (Vulpes vulpes) are the most abundant carnivore species in the Northern Hemisphere. Since their populations are well established in peri-urban and urban areas, they represent a potential reservoir of viruses that transmit from wildlife to humans or domestic animals. In this study, we evaluated the faecal virome of juvenile and adult foxes from peri-urban areas in central Croatia. The dominating mammalian viruses were fox picobirnavirus and parvovirus. The highest number of viral reads (N = 1412) was attributed to a new fox circovirus and complete viral genome was de novo assembled from the high-throughput sequencing data. Fox circovirus is highly similar to dog circoviruses identified in diseased dogs in USA and Italy, and to a recently discovered circovirus of foxes with neurologic disease from the United Kingdom. Our fox picobirnavirus was more closely related to the porcine and human picobirnaviruses than to known fox picobirnaviruses.     

gp64基因相应dsRNA对家蚕核型多角体病毒（BmNPV）增殖的抑制

 【目的】研究gp64基因相对应的多个dsRNA对家蚕核型多角体病毒增殖的抑制效果。【方法】体外合成dsRNA，通过病毒滴度试验、实时定量RT-PCR法研究gp64基因的不同区域、不同长度与RNAi干扰效果的关系。【结果】供试的6个dsRNA的最大抑制效果相当（滴度TCID50的最大抑制差值为4.00左右）；经RNAi的不同时间点的gp64 mRNA 表达水平均明显下调(P<0.01)，其中48 h的gp64 mRNA的表达量约为对照的1/300。【结论】6个dsRNA均能有效地抑制病毒的基因表达和增殖；gp64 ORF第1 390～1 499位点（G3-3）是RNAi的较佳靶位点。     

分泌抗BVDV McAb杂交瘤细胞株的建立

用牛病毒性腹泻病毒（ＢＶＤＶ）感染ＭＤＢＫ细胞,将感染细胞培养物冻融后离心取上汪经ＰＥＧ沉淀浓缩抗原免疫ＢＡＬＢ／Ｃ小鼠,取血清抗体效价高遥免疫鼠的脾细胞与ＳＰ２／Ｏ细胞在ＰＥＧ作用下融合,产生杂交瘤细胞,用间接ＥＬＩＳＡ法检测杂交瘤细胞生长孔上清,筛选阳性杂交瘤细胞株。以有限稀法克隆２－３次,得到８ 泌抗ＢＶＤＶ的单克隆抗体（ＭｃＡｂ）杂交瘤细胞株。８株ＭｃＡｂ对ＢＶＤＶ,猪瘟均呈阳性反应。杂交     

香菇病毒快速检测

应用十二烷基磺酸钠聚丙烯酰胺(SDS-PAGE)凝胶电泳分析香菇菌丝蛋白质,根据能否检出22 000道尔顿的34nm香菇病毒外壳蛋白带作为菌株是否带病毒的指标。在生长不正常香菇菌株和对照的带病毒菌株中均检测出了34nm香菇病毒外壳蛋白带。这一结果表明,应用SDS聚丙烯酰胺凝胶电泳可对香菇病毒进行简单、快速的早期检测。     

Neonatal Equine Herpesvirus Type 1 Infection on a Thoroughbred Breeding Farm

Of 17 foals born on a Thoroughbred breeding farm between March and April 1995, infection with equine herpesvirus type 1 (EHV-1) was associated with neonatal morbidity in 5 foals, 3 of which died or were euthanized. Morbidity and mortality were associated with pulmonary inflammation, and EHV-1 was identified in the lungs of the 3 foals that died. All neonatal EHV-1 infections occurred in foals of mares housed in the same pasture and barn. No other clinical manifestations of EHV-1 infection (eg, abortion, neurologic disease, or respiratory disease) occurred during this outbreak. Three foals were treated with acyclovir (1 died, 2 survived), which may have influenced the clinical outcome in the surviving foals.     

A comparative study of pigeons and chickens experimentally infected with PPMV-1 to determine antigenic relationships between PPMV-1 and NDV strains

To compare the pathogenicity of PPMV-1 in pigeons and chickens, both species of birds were experimentally infected with strain pi/CH/LHLJ/110822, which was isolated from a pigeon in China. The clinical signs, gross lesions, and histopathological changes were observed in pigeons inoculated with pi/CH/LHLJ/110822. The morbidity and mortality rates were 80% and 70% in pigeons, respectively, whereas there were no clinical signs or gross lesions in chickens inoculated with the same strain. The viral loads in tissue samples were detected by real-time RT-PCR, indicating that six tissue samples (i.e., kidney, lung, brain, trachea, Harderian glands, and proventriculus) had detectable viral RNA in all dead pigeons, and significant differences in viral loads between pigeons and chickens were observed in several tissue samples (i.e., Harderian glands, proventriculus, duodenum, pancreas, small intestine, and large intestine) on 3 days post-inoculation (dpi) and in brain tissue on 7 dpi. In general, viral loads in pigeons were higher than those in chickens, whereas antibody titers in pigeons were lower than those in chickens. These results showed differences in pathogenicity, efficiency of viral RNA replication, and humoral immunity, indicating different susceptibilities between the host species. Additionally, the cross hemagglutination inhibition assay and cross virus neutralization tests demonstrated that pi/CH/LHLJ/110822 antigenicity was different from those of strains La Sota and F48E9.     

真鲷幼鱼狂游症病毒病原学初步研究

应用电子显微镜观察真鲷幼鱼内脏组织超薄切片。发现真鲷幼鱼脾脏存在大量病毒颗粒。病毒颗粒直径约１２０ｎｍ，呈六角形，有一层蛋白质外壳。核心主要为核心酸。病毒颗粒主要位于细胞核内。感染的细胞肿胀，比正常细胞增大５－６倍。在肠组织超薄切片中也发现少量形态相同的病毒颗粒。     

全氟辛烷磺酸酶联免疫检测方法的研究

以人工合成的全氟辛烷磺酸免疫原免疫新西兰大白兔,制备多克隆抗体,用辛酸-硫酸铵沉淀法纯化抗体,选择包被抗原浓度为0.1μg/mL,抗体稀释度为1∶8 000,建立酶联免疫(ELISA)检测方法。交叉反应试验结果表明,除全氟癸烷磺酸和全氟己烷磺酸的交叉反应率偏高(16.2%,14.22%)外,与其他全氟化合物交叉反应率低于10%,即该抗体有较好的特异性。PFOS检测限为10μg/L,该ELISA方法的最佳工作范围在10~1 000μg/L之间,在此范围内平均回收率为101.62%,平均变异系数为2.4%,表明该方法准确度和精密度高,便于在线检测,可应用于对水体中全氟辛烷磺酸残留的酶联免疫检测。