PVY和PVS病毒复合侵染对马铃薯光合及营养品质的影响

以2个马铃薯高代品系为实验材料,研究马铃薯病毒Y(PVY)和S(PVS)复合侵染对马铃薯叶片SPAD值、最大净光合速率、表观量子效率、气孔导度、暗呼吸速率、蒸腾速率、光补偿点、胞间CO_2浓度和饱和光强等光合参数,以及马铃薯块茎产量、干物含量、淀粉含量、还原糖含量、粗蛋白含量等产量品质指标的影响。结果表明:(1)田间受到PVY和PVS病毒复合侵染后,马铃薯品系2010-11植株的叶片PVS病毒含量显著低于品系34-2植株,2010-11植株的叶片PVY病毒含量同样低于品系34-2植株,但差异不显著。(2)与对照相比,感病后马铃薯品系34-2和2010-11的平均单株产量分别显著降低48.54%和19.98%,淀粉含量分别降低8.06%和3.38%,但不显著;粗蛋白含量分别显著升高14.05%和29.17%;还原糖含量分别显著降低11.11%和14.29%;34-2干物量含量显著降低6.9%,2010-11干物质含量略降低0.66%。(3)两品系感病植株的光合参数SPAD值分别显著降低13.37%、20.10%;最大净光合速率分别显著降低32.48%和4.54%;其它光合参数变化没有规律性。研究发现,PVY和PVS病毒混合侵染使马铃薯植株叶片叶绿素含量显著降低,进而影响光合作用的正常进行;病毒侵染使块茎中还原糖和淀粉的积累显著受到抑制,但能够促进块茎中粗蛋白的含量显著提高。     

用SDS溶解的蛋白质的Dot—ELISA和免疫双扩散技术

有些难溶性蛋白质用非离子型去垢剂不能溶解,溶解这类蛋白质有时要用2％(w/v)十二烷基磺酸钠(SDS)和4％;(v/v)巯基乙醇。但SDS和巯基乙醇又能影响免疫反     

犬瘟热病毒（CDV）ELISA检测方法的建立与应用

目的建立犬CDV抗体ELISA检测方法。方法培养vero细胞,接种CDV病毒,制备vero正常抗原和CDV特异抗原,滴定酶结合物和抗原最佳工作浓度,并进行精密性、敏感性、稳定性、特异性实验。结果正常、特异抗原和酶结合物最佳工作浓度分别为1∶32 000、10μg/mL和1∶8 000;正常、特异抗原批内变异系数分别为9.1%和5.8%,批间平均变异系数分别为8.8%和6.6%;检测灵敏度为1∶2 560;与犬细小病毒（CPV）、犬肝炎病毒（ICHV）均无交叉反应。稳定性试验相对偏差小于25%。结论建立的ELISA方法重复性、稳定性好,特异性、敏感性强。可用于犬CDV抗体的检测。     

马铃薯Y病毒蚜传辅助成分介导PVX/PVY协生作用

构建了马铃薯Y病毒中国株系（PVY－C）蚜传辅助成分（HC－Pro）基因的正义、反义和缺失三种植物表达载体,通过农杆菌介导法转化烟草品种NC89。Southern blot分析表明,HC－Pro基因及其突变体已经整合到烟草染色体中,Western blot分析证明,正义HC－Pro基因及其缺失突变体在转基因烟草中有表达产物,攻毒试验结果表明,转正义,HC－Pro基因及其缺失突变体不仅能够提高T1转基因烟草中PVY－C的病毒积累和致病,而且对异源病毒PVX具有同样的作用,而转反义HC－Pro基因烟草对PVY－C和PVX的致病性无影响,因此,PVY－C HC－Pro基因介导PVX／PVY的协作作用。     

重庆烟草地区ELISA检测

为调查重庆主要烟草地区的病毒病种类,采用没脸免疫吸附法(ELISA)对采自重庆涪陵、彭水、万州、黔江、武隆、酉阳的258份的烟草样品进行了病毒病种类的检测.检测结果表明,其中274份样品呈阳性。主要受到7种病毒病的侵染,主要包括:烟草花叶病毒(Tobacco mosaic virus,TMV)、黄瓜花叶病毒(Cucumber mosaic virus,CMV)、马铃薯Y病毒(Potato virus Y,PVY)、马铃薯X病毒(Potato virus X,PVX)、芜菁花叶病毒(Turnip mosaic virus,TuMV)、蚕豆萎蔫病毒2号Broad bean wilt virus 2,BBWV-2)、烟草蚀纹病毒(Tobacco etch virus,TEV)。其中TMV的检出率最高,达50%,已成为重庆地区烟草优势病原。     

应用ELISA法检测SARS-Ab

探讨对SAPS病毒进行临床诊断,分析ELISA法检测SARS-Ab的情况。结果显示对照组检测均为阴性,测试组发病12d前、后阳性检出率分别为2．7％、92％,总检出率为94．7％。29例疑似病人阳性检出率为31％。提示该方法敏感性、特异性强,但12d以前检出率低,而13-16d以后阳性率明显提高。     

应用单克隆抗体ELISA检测淋球菌

利用抗淋球菌外膜蛋白的单克隆抗体（ＭｃＡｂ）制备的淋球菌酶免疫试剂（ＧＯＮＯ－ＥＩＡ）检测实验室已知淋球菌的阳性低限值菌数为６．２５×１０~４／ｍｌ。对脑膜炎奈瑟氏菌、微黄色奈瑟氏菌、卡它布兰汉氏球菌、大肠埃希氏菌、绿脓杆菌和乳酸杆菌等无交叉反应。检测临床生殖泌尿系统标本４８４例,以淋球菌培养法对比,ＧＯＮＯ－ＥＩＡ的敏感性为８１．４３％,特异性为９４．９２％,,总符合率为９２．９７％。ＧＯＮＯ－ＥＩＡ方法简便,快速,整个实验不超过１２０分钟,是适用于临床实验诊断的理想方法。     

Comparison of a Filter Paper Immunobinding Assay, Western Blotting and an Enzyme Linked Immunosorbent Assay for the Detection of Wheat Streak Mosaic Virus

The adaptation and improvement of serological assays for the detection of plant viruses has steadily developed. A filter paper immunobinding assay, Western blotting and the double sandwich ELISA were compared for the detection and quantification of wheat streak mosaic virus. ELISA was a more quantitative assay, but the filter paper immunobinding assay and Western blotting were more conservative of antiserum when only a few assays were required and could be run in a shorter period of time.     

植物组织粗汁液中的番木瓜环斑病毒的ELISA检测技术

本研究建立和改进了检测番木瓜和西葫芦组织粗汁液里的番木瓜环斑病毒（ＰＲＶ）的ＤＡＣ－ＥＬＩＳＡ法和Ｄｏｔ－ＥＬＩＳＡ法。用不同的ＥＬＩＳＡ方法来检测不同寄主植物粗汁液里的ＰＲＶ,其所用的合适的制备粗汁液的缓冲液是不同的。用ＤＡＣ－ＥＬＩＳＡ法检测西葫芦粗汁液时,以０．５ｍｏｌ／Ｌ磷酸盐缓冲液（ｐＨ７．５,内含０．１ｍｏｌ／Ｌ乙二胺四乙酸二钠）为宜;而检测番木瓜粗汁液时,则还要加入０．２５ｍｏｌ／Ｌ脲。用Ｄｏｔ－ＥＬＩＳＡ法检测时,在上述磷酸盐缓冲液中加入２％聚乙烯吡咯烷铜能提高对西葫芦粗汁液的检测效果。应用合适的制备粗汁液的缓冲液,ＤＡＣ－ＥＬＩＳＡ法和Ｄｏｔ－ＥＬＩＳＡ法的灵敏度分别提高到１／４０９６和１／１０２４（稀释度）。本研究还表明,影响ＤＡＣ－ＥＬＩＳＡ法的定过测定的主要因素是粗汁液的稀释度和包被液（０．０５ｍｏｌ／Ｌ碳酸盐缓冲液,ｐＨ９．６）的用过。在较高粗汁液稀释度和包被液的用量相同时,粗汁液里的病毒含量与ＤＡＣ－ＥＬＩＳＡ法的ＯＤ４９２ｎｍ值呈真实的线性关系。     

An Assay Method for Herpes Simplex Virus Type 1 Seroprevalence Survey—Detection of Antibody in Saliva by Avidin-Biotin Complex Enzyme-Linked Immunosorbent Assay (ABC-ELISA)

To determine whether the avidin-biotin complex enzyme-linked immunosorbent assay (ABC-E) is a potentially useful method for detection of herpes simplex virus type 1 (HSV-1) antibody in saliva, paired serum and saliva samples from 129 healthy individuals aged 18 to 25 years were collected simultaneously and subjected to a neutralization test (NT) for neutralizing antibody and also to an indirect ELISA (IE) and ABC-E for HSV-1 specific IgG detection. Compared with the results of NT, the sensitivities of the IE and ABC-E for serum were both 100% (45/45), and for saliva 82.2% (37/45) and 93.3% (42/45), respectively. The specificity of all these methods was 100% (84/84). With the same ABC-E method, a significant correlation (r=0.66, P < 0.001) between the OD-difference (d-OD) values of positive serum and saliva samples was observed. Furthermore, the consistency of ABC-E for salivary antibody detection was confirmed with the paired serum and saliva samples which were collected from four individuals followed up for eight months. It was clear that the ABC-E method for saliva can be used in place of the NT and ABC-E method for serum for seroprevalence studying of HSV-1 infection.     

Transfer of resistance to potato leafroll virus, potato virus Y and potato virus X from Solarium brevidens to S. tuberosum through symmetric and designed asymmetric somatic hybridisation

Resistance to potato leafroll virus (PLRV), potato virus Y (PVY^o) and potato virus X (PVX) was studied in symmetric and asymmetric somatic hybrids produced by electrofusion between Solanum brevidens (2n=2×=24) and dihaploid S. tuberosum (2n=2×=24), and also in regenerants (B-hybrids) derived through protoplast culture from a single somatic hybrid (chromosome number 48). All of the somatic hybrids between 5. brevidens and the two dihaploid lines of potato cv. Pito were extremely resistant to PLRV and PVY^oand moderately resistant to PVX, irrespective of their chromosome number and ploidy level (tetraploid or hexaploid). Most (56%) of the asymmetric hybrids of irradiated S. brevidens and the dihaploid line of potato cv. Pentland Crown (PDH40) had high titres of PVY^osimilar to those of PDH40, whereas the rest of the hybrids had PVY^otitres less than a tenth of those in PDH40. Three B-hybrids had a highly reduced chromosome number (27, 30 and 34), but were however as resistant to PLRV, PVY^oand PVX as 5. brevidens. Two asymmetric hybrids and one B-hybrid were extremely resistant to PLRV but susceptible to both PVY and PVX. The results suggested that resistance to PLRV in 5. brevidens is controlled by a gene or genes different from those controlling resistance to PVY and PVX, and the gene(s) for resistance to PVY and PVX are linked in S. brevidens.     

Detection of Potato Viruses Y and A in Tubers by Enzyme-Linked Immunosorbent Assay after Natural and Artificial Break of Dormancy

The effects of natural and artificial break of dormancy on the detection of potato viruses Y (PVY) and A (PVA) in tubers of field-grown potato cultivars with primary and secondary infections were studied by enzyme-linked immunosorbent assay (ELISA). Both viruses were at low concentration and unevenly distributed in dormant -tubers. Four to six weeks after treatment with Rindite, maximum concentrations of PVY and PVA were detected at both tuber ends with generally higher values at the rose end than at the heel end. Slower and smaller virus increase and a retarded and/or limited virus translocation were observed in treated tubers of resistant cultivars. Consequently, detectability of PVY and PVA was higher in tubers of susceptible cultivars than in those of the resistant ones. Detection of both viruses was very difficult and unreliable in tubers after natural break of dormancy. The very low concentration of PVY and PVA in dormant tubers and their drastic multiplication in Rindite- treated tubers appear to be characteristic of potyviruses and are unique among the major potato viruses.