三带喙库蚊体内猪繁殖与呼吸综合征病毒的分离与鉴定

【目的】调查猪场蚊虫是否能携带猪繁殖与呼吸综合征(PRRS)病毒。【方法】采集发生PRRS疫情的3个养猪场蚊虫样本,采用RT-PCR方法检测PRRS病毒核酸,取阳性蚊虫样本接种Marc-145细胞进行病毒的分离培养,以间接免疫荧光抗体技术和分子克隆技术进行病毒的鉴定。【结果】 养猪场内的蚊虫主要有三带喙库蚊Culex tritaeniorhychus、凶小库蚊Culex modestus、中华按蚊Anopheles sinensis和骚扰阿蚊Armigeres obturbans,其中三带喙库蚊占86.76%;以PRRS病毒N基因引物进行扩增,三带喙库蚊样本呈现阳性反应,而其他蚊种均为阴性。在蚊虫接种的Marc-145细胞中可见细胞融合和空泡形成等细胞病变效应;用抗PRRS病毒N蛋白抗体和羊抗猪IgG(H+L)-FITC进行间接免疫荧光染色,感染细胞呈现黄绿色荧光;以NSP2基因引物进行RT-PCR扩增、克隆与测序,发现库蚊源病毒与相应猪场猪源病毒中相应基因的序列具有较高同源性。【结论】 三带喙库蚊为猪舍优势蚊种,并能携带猪繁殖与呼吸综合征病毒。     

猪繁殖与呼吸综合征疫苗

针对猪繁殖与呼吸综合征(porcine reproductive and respiratory syndrome,PRRS),主要以疫苗预防为主.常规疫苗主要有灭活疫苗和弱毒疫苗,但前者的免疫原性差,保护效果不佳;后者又发现存在毒力返强的可能性,安全性低的问题.目前学者们主要集中在对猪繁殖与呼吸综合征病毒(PRRS virus,PRRSV)基因工程疫苗进行研究.本文就PRRS的灭活疫苗、弱毒疫苗和基因工程疫苗的研究进展做详细的阐述.     

利用RNA干扰机制抑制猪繁殖与呼吸综合征病毒的增殖

本文探讨依靠RNAi技术对猪繁殖与呼吸综合征病毒(PRRSV)增殖的干扰作用。筛选到针对编码PRRS病毒核衣壳蛋白的N基因的两处靶序列作为候选片段,在MARC-145细胞上进行基因干扰实验研究。成功观测到由载体表达的小干扰RNA(siRNA)在MARC-145细胞中对PRRS病毒增殖的抑制现象。通过选取不同时间段对病毒进行TCID50检测,以及对CPE出现时间进行观察和免疫荧光技术,得到RNA干扰对PRRS病毒增殖抑制作用的动态数据。证实在真核细胞水平上,RNA干扰机制可以抑制PRRS病毒的增殖。实验结果表明,依靠载体表达的RNA干扰技术将会对今后针对PRRS病毒的新型疫苗开发提供一个新思路。     

利用RNA干扰机制抑制猪繁殖与呼吸综合征病毒的增殖

本文探讨依靠RNAi技术对猪繁殖与呼吸综合征病毒( PRRSV) 增殖的干扰作用.筛选到针对编码PRRS病毒核衣壳蛋白的N基因的两处靶序列作为候选片段,在MARC-145细胞上进行基因干扰实验研究.成功观测到由载体表达的小干扰RNA (siRNA)在MARC-145细胞中对PRRS病毒增殖的抑制现象.通过选取不同时间段对病毒进行TCID50检测,以及对CPE出现时间进行观察和免疫荧光技术,得到RNA干扰对PRRS病毒增殖抑制作用的动态数据.证实在真核细胞水平上,RNA干扰机制可以抑制PRRS病毒的增殖.实验结果表明,依靠载体表达的RNA干扰技术将会对今后针对PRRS病毒的新型疫苗开发提供一个新思路.     

融合表达牛疱疹病毒1型VP22及猪繁殖与呼吸综合征病毒GP5重组伪狂犬病毒TK-/gE-/VP22GP5+的构建

猪繁殖与呼吸综合征(porcine reproductive andrespiratory syndrome,PRRS)和伪狂犬病(pseudora-bies,PR)是两种严重危害养猪业的重要病毒性传染病。而引发PR的伪狂犬病病毒(pseudorabiesvirus,PRV)是构建基因工程疫苗优良的活病毒载体[1]。已有报道将表达猪瘟病毒(hog cholera virus,HCV)囊膜蛋白E1基因的重组PRV(rPRV)二价基因工程疫苗免疫猪后,能同时抵抗HCV和PRV的强毒攻击[2],显示了PRV作为活病毒载体的可行性和“一针防两病”的独特优点。由ORF5基因编码的囊膜糖蛋白GP5是猪繁殖与呼吸综合征病毒(porcine reproductive …     

猪繁殖与呼吸综合征病毒实时PCR检测方法的建立

设计合成了一套引物和TaqMan探针,以扩增猪繁殖与呼吸综合征病毒的核衣壳蛋白基因,通过反应条件的优化,在国内首次建立了快速定量检测猪繁殖与呼吸综合征病毒的实时PCR方法,并用该法检测患病猪的肺脏等样品。结果表明该方法具有较好的特异性和重复性,对PRRSV细胞培养物的检测下限为0.01TCID50,敏感性比常规RT-PCR高100倍;对10份PRRS疑似猪肺脏样品检测5份为阳性,与病毒分离的阳性符合率为100%。该方法具有快速、灵敏、准确、低污染等优点,在PRRSV的早期检测、预防控制、进出口检疫及基础研究中会起到重要作用。     

应用RT-PCR检测流产胎儿组织中猪繁殖与呼吸综合征病毒

根据猪繁殖与呼吸综合征病毒(PRRSV)美洲型膜蛋白和核衣壳蛋白基因序列,设计了一对含有EcoR I和BamH I酶切位点的引物,用RT-PCR对四个流产猪场的病料进行了检测,扩增出约918 bp的基因片段.通过病毒分离、酶切鉴定和序列分析证实为PRRSV感染.结果说明应用所设计的引物进行RT-PCR快速检测PRRS是可行的,为我国快速特异诊断PRRS和PRRSV强毒株的深入研究奠定了基础.     

猪繁殖与呼吸综合征病毒实时PCR检测方法的建立

设计合成了一套引物和TaqMan探针,以扩增猪繁殖与呼吸综合征病毒的核衣壳蛋白基因,通过反应条件的优化,在国内首次建立了快速定量检测猪繁殖与呼吸综合征病毒的实时PCR方法,并用该法检测患病猪的肺脏等样品.结果表明该方法具有较好的特异性和重复性,对PRRSV细胞培养物的检测下限为0.01TCID50,敏感性比常规RT-PCR高100倍;对10份PRRS疑似猪肺脏样品检测5份为阳性,与病毒分离的阳性符合率为100%.该方法具有快速、灵敏、准确、低污染等优点,在PRRSV的早期检测、预防控制、进出口检疫及基础研究中会起到重要作用.     

不怕蓝耳病的基因编辑猪

猪呼吸与繁殖综合征(porcine reproductive and respiratory syndrome, PRRS)是PRRS病毒引发的一种严重危害养猪业的传染病,具有高变异性、持续性感染和免疫抑制等生物学特性。因病猪具有耳部变蓝的症状被俗称为"蓝耳病",又被称为养猪业中的"艾滋病"。猪肺泡巨噬细胞上的CD163蛋白是猪呼吸与繁殖综合征病毒的主要受体,研究者利用CRISPR/Cas9基因编辑技术敲除猪受精卵中的CD163基因第七外显子,成功制备出具有猪呼吸与繁殖综合征抗性的活体猪。这是抗猪呼吸与繁殖综合征的相关工作中一项里程碑式的研究成果,为该病未来的防治工作提供了新的思路和广阔的应用前景。     

猪繁殖与呼吸综合征病毒N基因和流感病毒HA基因的联合表达及应用

参照已公布的流感病毒血凝素基因(HA基因)及猪繁殖与呼吸综合征病毒(PRRSV)基因组序列,设计并合成一对引物P1、P2,以RT-PCR方法扩增出PRRSV的ORF7片段(约410bp),其中含HA基因主要核苷酸序列(33bp).用BamH I、Xho I分别对扩增出的片段及pET32a质粒进行酶切,连接后构建了重组质粒pETHN并转化到BL21(DE3)宿主菌中诱导表达.用纯化后的表达产物与流感病毒血凝素单抗及乳胶建立了诊断猪繁殖与呼吸综合征(PRRS)的乳胶凝集试验.检测结果显示该方法有良好的特异性及敏感性,与IDEXX公司ELISA检测试剂盒符合率达93.8%.     

Proteomics in reproductive biology: Beacon for unraveling the molecular complexities

Proteomics, an interface of rapidly evolving advances in physics and biology, is rapidly developing and expanding its potential applications to molecular and cellular biology. Application of proteomics tools has contributed towards identification of relevant protein biomarkers that can potentially change the strategies for early diagnosis and treatment of several diseases. The emergence of powerful mass spectrometry-based proteomics technique has added a new dimension to the field of medical research in liver, heart diseases and certain forms of cancer. Most proteomics tools are also being used to study physiological and pathological events related to reproductive biology. There have been attempts to generate the proteomes of testes, sperm, seminal fluid, epididymis, oocyte, and endometrium from reproductive disease patients. Here, we have reviewed proteomics based investigations in humans over the last decade, which focus on delineating the mechanism underlying various reproductive events such as spermatogenesis, oogenesis, endometriosis, polycystic ovary syndrome, embryo development. The challenge is to harness new technologies like 2-DE, DIGE, MALDI-MS, SELDI-MS, MUDPIT, LC–MS etc., to a greater extent to develop widely applicable clinical tools in understanding molecular aspects of reproduction both in health and disease.     

猪繁殖与呼吸综合征检测技术研究进展

猪繁殖与呼吸综合征是导致猪繁殖障碍疾病的一种重要猪病。其致病机制和自身复制规律至今尚不完全明确,因此无法从根本上清除该病的存在,这就使得对该病的检测显得尤为重要。现从直观的临床诊断、精确的实验室诊断及鉴别诊断等方面对猪繁殖与呼吸综合征检测技术进行了综述,其中对可以最终确诊的实验室检测技术进行了重点介绍,并对猪繁殖与呼吸综合征生物学检测技术进行了总结。     

The C/EBPβ-Dependent Induction of TFDP2 Facilitates Porcine Reproductive and Respiratory Syndrome Virus Proliferation

Virologica Sinica - Porcine reproductive and respiratory syndrome (PRRS) is an important infectious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), leading to...     

Porcine reproductive and respiratory syndrome virus

Porcine reproductive and respiratory disease (PRRS) is an economically important disease around the globe; it has been estimated to cost the swine industry in USA approximately 560 million US dollars annually. It is well established that PRRS is caused by an enveloped, single-stranded positive-sense RNA virus known as porcine reproductive and respiratory syndrome virus (PRRSV). The inability to successfully control PRRS across farms via traditional methods (e.g. vaccine and animal flow) has led to a growing interest in area-based eradication. Important to such an initiative is information on PRRSV transmission within and between herds and intervention strategies to prevent its spread. This paper will review the current literature on selected areas of PRRS known to be important to the topic of pathogen elimination, including etiology, clinical manifestations, direct and indirect routes of transmission, as well as discuss measures for disease control, prevention and eradication.     

斑马鱼繁殖内分泌学研究进展

作为四大模式动物之一,斑马鱼(Danio rerio)广泛应用于胚胎学、发育生物学、毒理学、分子生物学等研究。但关于斑马鱼繁殖内分泌生理和环境毒理方面的研究少见报道,本文综述了近年来,在内分泌学方面以斑马鱼作为实验动物的研究概况。     

Molecular markers: progress and prospects for understanding reproductive ecology in elasmobranchs

Application of modern molecular tools is expanding the understanding of elasmobranch reproductive ecology. High-resolution molecular markers provide information at scales ranging from the identification of reproductively isolated populations in sympatry (i.e. cryptic species) to the relationships among parents, offspring and siblings. This avenue of study has not only augmented the current understanding of the reproductive biology of elasmobranchs but has also provided novel insights that could not be obtained through experimental or observational techniques. Sharing of genetic polymorphisms across ocean basins indicates that for some species there may be gene flow on global scales. The presence, however, of morphologically similar but genetically distinct entities in sympatry suggests that reproductive isolation can occur with minimal morphological differentiation. This review discusses the recent findings in elasmobranch reproductive biology like philopatry, hybridization and polyandry while highlighting important molecular and analytical techniques. Furthermore, the review examines gaps in current knowledge and discusses how new technologies may be applied to further the understanding of elasmobranch reproductive ecology.     

新型冠状病毒与系统损伤研究

自2019年12月全国及世界范围爆发了新型冠状病毒性肺炎(corona virus disease 2019,COVID-19),给中国和全球公共卫生安全带来了极大的挑战。研究发现,新型冠状病毒(severe acute respiratory syndrome coronavirus 2,SARS-CoV-2)不仅损伤呼吸系统,对心血管系统、泌尿系统、消化系统、中枢神经系统、免疫系统和生殖系统等均有不同程度损伤。因此,全面掌握SARS-CoV-2的基本情况、COVID-19的流行病学特征及引发的主要系统损伤,可为COVID-19的筛查、诊断、防治及预后提供依据。     

猪繁殖与呼吸综合征病毒分子生物学研究进展

猪繁殖与呼吸综合征病毒(PRRSV)是严重危害养猪业的病原,对PRRSV的分子生物学研究进展进行了综述,主要包括PRRSV的基因组结构、病毒的非结构蛋白和结构蛋白及其功能、致病机理及复制与转录等.     

Developmental biology meets with reproductive engineering; interdisciplinary science area as a breakthrough.

It has been postulated many times that different scientific topics and strategies often encounter each other, which create cutting edge research field resulting in further significant progresses of science. This was also a lesson bestowed by Prof. Raymond Wegmann where he created innovative research field for biology, molecular biology and biochemistry-biophysics. Progresses of developmental biology were boosted by molecular biology and reproductive engineering where ES cells and embryonic manipulation are necessary. There are no questions about the utility of their technologies. Reviews on their contributions with respect to the condition of genome manipulation are addressed.