新型HCV EIA诊断试剂盒的研制

丙型肝炎病毒（ＨＣＶ）基因组结构区核壳蛋白（Ｃ）区抗原、膜蛋白Ｅ１和Ｅ２区抗原,以及非结构区ＮＳ３－ＮＳ５区抗原的区段,已经在原核细胞中获得有效的表达。同时,相应区段中的优势抗原表位肽也经化学合成法大规模地制备。ＨＣＶ基因组上各区段抗原性的分析发现,由Ｃ区和ＮＳ３区分别编码的Ｃ抗原和Ｃ３３ｃ抗原是ＨＣＶ基因组上两个优势抗原区段。其相应的抗体出现早（感染后６周可检出抗Ｃ３３ｃ抗体）,阳转率高（约９９％阳性检出率）,特异性和重复性均优于其它区段抗原。以中国人ＨＣＶ的Ｃ３３ｃ重组蛋白和分支状合成肽ＭＡＰ－Ｃ－１９为复合抗原,研制了适合我国抗ＨＣＶ抗体检测的新型丙型肝炎病毒酶免疫测定（ＨＣＶＥＬＡ）诊断试剂盒。它同当代美国Ａｂｂｏｔｔ／ＵＢＩＨＣＶＥＬＡ诊断试剂的符合率约９８％,同加拿大ＹＥＳ公司ＨＣＶＥＩＡ诊断试剂的符合率约９７．８％,阳性检出率提高了约２％,３次重复性达１００％,表明其特异性、敏感性和重复性均达到了当代第二代ＪＣＶＥＬＡ诊断试剂的水平。我国人群中抗ＨＣＶ抗体的分布情况为：正常人群的检出率１％－２％;外科类住院病人检出率约２８．８％;肝炎患者抗ＨＣＶ阳性率为３４．４％,慢活肝、肝硬化和重症肝炎患者     

抗MRSA-PBP2a鸡卵黄抗体的制备及乳胶凝集检测法的建立

制备抗耐甲氧西林金黄色葡萄球菌青霉素结合蛋白2a( MRSA- PBP2a)抗原的鸡卵黄免疫球蛋白(IgY),建立检测MRSA的乳胶凝集方法.采用体外诱导的方法制备PBP2a蛋白,胸部肌肉多点注射方式免疫6只海蓝蛋鸡,水稀释法提取IgY,BCA法测定蛋白含量,Western blotting进行特异性分析,用提取的IgY抗体致敏聚苯乙烯乳胶,建立检测PBP2a的乳胶凝集方法.成功诱导并制备获得纯化的PBP2a蛋白,首次免疫后1月每枚鸡蛋提纯后可获得约48 mg IgY抗体,Western blotting结果显示IgY抗体能有效识别纯化的PBP2a蛋白;成功建立检测PBP2a的乳胶凝集法,敏感性达1 mg/L.抗MRSA- PBP2a鸡卵黄抗体具有较高的敏感性和特异性,基于其建立的乳胶凝集检测方法具有较好的灵敏性.     

动物组织中磺胺二甲嘧啶残留ELISA试剂盒研制

采用重氮化法和戊二醛法,将磺胺二甲嘧啶分别与牛血清白蛋白和辣根过氧化物酶偶联制备了免疫原和酶标半抗原,免疫兔获得了特异性抗体,成功建立了相关动物产品中磺胺二甲嘧啶残留ELISA定量检测方法及商品化试剂盒,并对试剂盒的灵敏度、准确度、精密度和稳定性进行了研究。试剂盒检测线性范围为62.5~0.54 ng/mL。在待测样品中各添加500、200、100、50 ng/g SMZ,测试的回收率平均为89.0%~134.8%;试剂盒测定结果与色谱的平均符合率99.8%~126.0%;对比定性测试15份色谱检测为阴性的样品,均未出现假阳性。试剂盒存放在37℃10 d和2~8℃5个月,质量稳定。     

Ⅱ型猪圆环病毒检测试剂盒的研制

猪圆环病毒(Porcine circovirus,PCV)属圆环病毒科圆环病毒属成员。PCV分两种血清型,即PCV-1和PCV-2,其中PCV-1由Tischer等于1974年检测到,对猪没有致病性;PCV-2可引起部分断奶仔猪和育肥猪的断奶仔猪多系统综合征(postweaning multisystemic wasting syndrome,     

牛血清蛋白酶联免疫检测试剂盒的研制及验证

为研制酶联免疫试剂盒以检测病毒性疫苗中残余牛血清蛋白(BSP)含量,制备高效价高纯度的兔抗BSP多克隆抗体作为包被抗体和酶标抗体,建立了ELISA双抗体夹心法并组建试剂盒,通过标准剂量曲线可对样品中所含BSP、BSA及B-IgG进行定量,经验证该方法标准曲线线性范围内r≥0.98,对BSP的检测限量为3ng/ml;分别检测5、10、20ng/ml含量的BSP时,试验内(n=12)和试验间(n=3)测定的变异系数在3.71%到7.29%之间,回收率在93.4%~106.3%,未见该方法与人血清白蛋白、卵清蛋白以及疫苗复合保护剂之间有交叉反应。该法敏感度高,准确性、重复性和稳定性好,可用于疫苗牛血清残余蛋白的质量控制。     

II型猪圆环病毒检测试剂盒的研制

猪圆环病毒(Porcine circovirus, PCV)属圆环病毒科圆环病毒属成员[1]。PCV分两种血清型,即PCV 1 和 PCV 2,其中 PCV 1 由 Tischer 等[2] 于1974年检测到,对猪没有致病性;PCV 2 可引起部分断奶仔猪和育肥猪的断奶仔猪多系统综合征( postweaning multisystemic wasting syndrome,PMWS)[3]。目前,该病呈世界分布,给养猪业造成巨大的经济损失。本研究通过分析已发表的 PCV 2的基因序列,找出 PCV 2 的特异性片段,研制了检测PCV 2的试剂盒。检测临床疑似病料,结果表明该试剂盒使用方便、快速、敏感、特异,符合国内的养殖场…     

细菌生理特性快速检测试剂盒研制

选取各代谢途径中利用某些特定碳源,如蔗糖、葡萄糖、甘露醇等95种碳源作为测试碳源,分别加入96孔酶标板中.选用指示剂噻唑兰(MTT)来鉴定细菌利用特征碳源情况.同时选取不同种类和不同浓度的抗生素,不同起始pH、不同NaCl浓度加入96孔酶标板中,测定细胞是否生长,可以快速地获得该菌的抗生素耐受性谱.试验还从指示剂浓度、培养基的固化、接菌浓度、观察时间等方面对试剂盒进行优化,得到一种最佳的试剂盒鉴定系统BiobiqA(碳源利用谱)和BiobiqB(生理抗性谱).用模式菌株对试剂盒进行测试和验证,结果表明,试剂盒具有操作简便、结果准确、节省成本、节约时间等优点,可以进行细菌生理特性的快速检测.     

Ⅱ型猪圆环病毒检测试剂盒的研制

猪圆环病毒(Porcine circovirus,PCV)属圆环病毒科圆环病毒属成员[1].PCV分两种血清型,即PCV-1和PCV-2,其中PCV-1由Tischer等[2]于1974年检测到,对猪没有致病性;PCV-2可引起部分断奶仔猪和育肥猪的断奶仔猪多系统综合征(postweaning multisystemic wasting syndrome,PMWS)[3].目前,该病呈世界分布,给养猪业造成巨大的经济损失.本研究通过分析已发表的PCV-2的基因序列,找出PCV-2的特异性片段,研制了检测PCV-2的试剂盒.检测临床疑似病料,结果表明该试剂盒使用方便、快速、敏感、特异,符合国内的养殖场及进出口的需要.     

小儿副轮状病毒的检出

用聚丙烯酰胺凝胶电泳法,从婴幼儿急性腹泻粪便中检出轮状病毒RNA电泳阳性293份,发现一株副轮状病毒(青-27株),此株病毒经电泳观察,呈典型的轮状病毒形态,但易破碎。ELISA检测表明不具有一般轮状病毒的特异性群抗原,病毒RNA基因组由11个片段组成,但电泳图型特殊,吴4:3:2:2排列模式,本文证实,此一小儿副轮状病毒与国外报道的,散发罕见的小儿副轮状病毒RNA电泳图型相同,提示此病毒的重要意义。     

Overexpression of l-galactono-1, 4-lactone dehydrogenase (GLDH) in Lanzhou lily (Lilium davidii var. unicolor) via particle bombardment-mediated transformation

In China, the native Lanzhou lily (Lilium davidii var. unicolor) is used as an edible lily and is an important source of ascorbic acid. However, an efficient transformation system has not been developed for this important crop. Here, we describe the first successful genetic transformation mediated by particle bombardment in this lily variety. We used pCAMBIA-GLDH, which contains the l -galactono-1, 4-lactone dehydrogenase gene from apple, a hygromycin phosphotransferase gene as a selectable marker, and an intron-containing β-glucuronidase gene as a reporter. In all, 35 transgenic lines were selected via GUS assays and PCR; from these, eight were confirmed by Southern analyses. Transgenic plants were transferred to soil and grown under greenhouse conditions. The levels of reduced ascorbate and total ascorbate (reduced ascorbate + oxidized ascorbate) in transgenic plants were found to be two to seven times those in non-transgenic controls.     

人呼吸道合胞病毒融合蛋白(F)片段原核表达、纯化和鉴定

目的克隆并表达人呼吸道合胞病毒(Human respiratory syncytial virus,HRSV)兰州株的融合蛋白(F)基因片段。方法利用PCR技术扩增HRSV兰州株的融合蛋白基因片段,克隆于原核表达载体pET-42b(+),转化大肠杆菌(Rosetta),经IPTG诱导表达,镍离子亲和层析柱纯化,SDS-PAGE和Western-blot分析重组蛋白的表达及其反应原性。结果 PCR扩增得到951 bp的DNA片段,重组质粒pET42b-F经酶切鉴定和测序分析,表明质粒构建正确。表达的重组蛋白的相对分子质量为68 710,表达的重组蛋白占总菌体蛋白的7%,纯化后蛋白纯度达80%。经Western-blot分析,重组蛋白与抗RSV的单抗呈专一性强阳性反应。结论成功构建了HRSV兰州株F基因片段原核表达载体,并在大肠杆菌Rosetta中获得了表达,表达的重组蛋白具有反应原性和特异性,为HRSV感染引起的疾病血清学诊断以及试剂盒的研发提供了材料。     

The 2nd IALE Asia-Pacific Region Conference Landscape Change and Human Activity Lanzhou, China, Sept. 22 - 25th, 2001 (Call for papers, first round announcement)

Jointly orgagnized by:IALE- China; IALE-Australia Cold and Arid Regions Environmental & Engineering Research Institute, CASProgramme Committee Richard Hobhs, Perth, Australia; Xiao Duning, Shenyang, China;Nobukazu Nakagoshi, Hiroshima, Japan; Sun- Kee Hong, Seoul,Korea; Parida Kuneepong, Bangkok, Thailand; Elena Klimina,Khabarovsk, Russia; Kalpana Bhakuni, India; Zhan Zhiyong, Hong Kong, China; Chun-Yen Chang, Taibei, China Organizing Committee Xiao Duning, Institute of Applied Ecology, CAS, China(Chair person)Richard Hobbs, Murdoch University, Australia (Chair person)Cheng Guodong, Cold and Arid Regions Environmental & Engineering Research Institute, CAS, China Fu Bojie, Research Center for Eeo- environmental Sciences, CAS,China Cui Haiting, Beijing University, China Yang Yiguang, Yunnan University, China Wang Gang, The State Key Laboratory of Arid Agriculture, Lanzhou University, China.Objectives and Topics The aim of the conference is to help participants to approach real landscape change and human activity impact in Asia - Pacific region, including restoration and reconstruction of landscape ecosystem both locally and regionally. A. Theoretical aspects and quantitative approaches: Landscape pattern and processes; Landscape modeling; GIS and Remote Sensing. B. Landscape change and driving forces: Monitoring of landscape changes; Human activity and land use;Culture and landscape. C. Landscape management and reconstruction: Ecological restoration of natural areas;Landscape planning in rural areas; Ecological risk and security assessment for catchments; Ecological engineering in agricultural landscapes. D. Environment protection and ecological development in Western China: Management of oasis landscape;Desert landscape and desertification control;Biodiversity conservation; Landscape Resonrces and tourism. E. Urban landscape ecology.Preliminary conference program - September 22, arrival in Lanzhou, Registration. - September 23, plenary sessions- September 24, in-conference excursion (Lanzhou city and suburbs)- September 25, parallel sessionsWorking language: English Post conference excursions September 26 - 29-No. 1 Silk Road and Arid Region Landscape: Desert, Oasis, and Ancient Culture Heritage (Dunhuang).- No.2 Qinghai- Xizang(Tibetsn) Plateau Landscape: High-Cool Meadow, Salt Lake and Qinghai Lake, Cool Desert - No.3 World Natural Heritege: Jiuzhaigou, Sichuan Province (Forest and Waterfall)Call for papers:Participants intending to present a paper or a poster during the conference are requested to send an abstract (no longer than 500 words) as email or TXT format attachment to the conference secretary (Landscape2001@sina.com). Or, you can send the hard copy to: Dr. CAO Yu, Institute of Applied Ecology, CAS, P. O. Box 417, 110016,Shenyang, China. Please include the participant's full address (postal,phone, fax, email) in the abstract. The deadline for abstract submission is June 30th, 2001. Abstracts will be published as conference proceedings, which will be available at the conference. Selected papers will be published after the conference.Reglstration:Please notify the conference secretary about your interest for participation. Further conference announcement and registration forms will be sent to you later soon. Registration fee for the conference: US＄ 300, including all meals during the conference, and coffee break refreshments. Room rent will be on your own. Registration fee for the excursions: US＄ 500, including transportation and accommodations.Contact address:Academic information (Abstract etc.): Dr. LI Xiuzhen; Dr. CAO Yu (Institute of Applied Ecology CAS, Shenyang, 110016, China)Tel: 86 - 24 - 23916291, Fax: 86 - 24 - 23843313.General information: Pro. Wang Genxu; Dr. Jiao Yuanmei (Cold and Arid Regions Environmental & Engineering Research Institute, CAS,Lanzhou, 730000, China) Tel: 86 - 931 - 8275120, 8275122,Fax: 86 - 931 - 8273894.Email: Landscape2001@sina. con     

On the Knowledge of Coprophilus (Zonyptilus) pennifer (Motschulsky, 1845) and Coprophilus (Zonyptilus) marginalis (Reitter, 1894) (Coleoptera,Staphylinidae)

Entomological Review - The type and other extensive material of Coprophilus (Zonyptilus) pennifer (Motschulsky, 1845) and some of its synonyms was examined. Coprophilus (Zonyptilus) marginalis...     

On the Synonymy of Coprophilus (Zonyptilus) pennifer (Motschulsky, 1845) (Coleoptera,Staphylinidae, Oxytelinae)

Entomological Review - The type material of several synonyms of Coprophilus (Zonyptilus) pennifer (Motschulsky, 1845) is revised. Both Coprophilus (Zonyptilus) striatipennis (Eppelsheim, 1894) and...     

Genetic polymorphism (ABO, Rh, Kell) in Kabul (Afghanistan)

ABO, rhesus and Kell blood group data on 1327 donors in Kabul are analysed by ethnic affinity and compared with existing data on Afghanistan peoples. Blood group frequencies are very similar in Pushtu and Tadjik, despite their different historical, linguistic, and cultural backgrounds. Inclusion of the small sample of Hazara in the analysis shows overall heterogeneity in rhesus D, E, and e frequencies, suggesting the existence of a broader pattern of genetic variation among the peoples of Afghanistan.     

Spermiogenesis in Anaspides tasmaniae (Thomson) (Crustacea,Malacostraca, Syncarida)

Spermiogenesis of the syncarid Anaspides tasmaniae (subclass Eumalacostraca) was investigated by transmission electron microscopy. The spermatozoan of Anaspides is an ovoid cell with an acrosome covering the anterior pole and a lobulated nucleus and mitochondria occupying the rest of the cell. A long subacrosomal filament bypasses the nucleus and forms a spiral that supports a thin extension of the posterior cytoplasm, giving the spermatozoan a bell-shaped appearance. No flagellum is present at any stage. The immobile spermatozoans are embedded in a hard capsule, secreted by the cells of the wall of the vas deferens.     

Tachinid (Diptera, Tachinidae) parasitoids of sawflies (Hymenoptera, Symphyta)

A list of 31 genera of tachinid parasitoids of sawflies is given. Despite the ecological and habitual similarity of sawflies and Lepidoptera, utilization of sawflies by tachinids was not as successful from the evolutionary point of view as utilization of Lepidoptera. Parasitism of tachinids in sawflies is evidently secondary; it appeared repeatedly and originates from parasitism in Lepidoptera. Most genera of tachinids are polyphagous. Endoparasitic Hymenoptera are koinobionts similarly to tachinids. The direct contact of the parasitoid with active mobile host larva not only results in complication of behavior and its specialization, but also promotes formation of different morphological innovations faster than that in idiobionts.