小麦苗枯病菌的ITS分析及PCR检测

 小麦苗枯病菌(Clavibacter fangii,Cf)是引起小麦细菌性苗枯病的病原,本研究用16S~23S rDNA间的内源转录间隔区(internally transcribed spacer,ITS)序列通用引物L1(5'-AGTCGTAACAAGGTAGCCGT-3')和L2(5'-GTGCCAAGGCATCCACC-3')扩增Cf和其它相关细菌的基因组DNA;并对其PCR产物进行回收、克隆和测序,将所获序列和其它已报道的细菌ITS序列进行多重比较后设计出Cf的特异性引物I1(5'-TGCCAAGTCACACTGAGACGA-3')和I2(5'-CAATGATCTACCACCCTCCGA-3')。此引物可以从Cf中扩增出351bp的特异性片段,而其余参试的21个细菌PCR反应结果均为阴性。该方法可以应用于小麦苗枯病菌的快速、可靠检测。此外,本研究对多种植物病原棒形杆菌的ITS序列进行比较研究,发现其具有一定的分类意义。     

利用RT-PCR检测库尔勒香梨苹果茎痘病毒的研究

 以库尔勒香梨新鲜、冷藏、冷冻叶片和皮层为材料,对提取双链RNA (dsRNA)的2种方法和提取总RNA的3种方法进行了分析比较,并对总RNA的提取方法进行了改进,获得了纯度较高、完整性较好的dsRNA和总RNA,在此基础上进行了反转录(RT)和PCR扩增。在国内首次完成了对苹果茎痘病毒(ASPV)的RT-PCR检测,建立了ASPV有效RT-PCR反应体系。用此体系扩增到ASPV一个长约316 bp的片段。实验表明以dsRNA和总RNA为模板均能成功进行RT-PCR检测,且dsRNA优于总RNA。     

不同引物对植原体的检测灵敏度比较研究

用常规PCR和巢式PCR对目前常用的各种植原体引物(fPl/rP7、fU5/rP7、fU5／rU3、fAY/rEY和fFD9/rFD9)的检测灵敏度进行了比较研究。研究发现，它们的检测灵敏度并不相同。最灵敏的引物(fAY/rEY)要比最不灵敏的引物(fFD9/rFD9)的灵敏度至少高出数千倍。因此，按照不同的检测目的选择所用的检测引物是明智的，特别是当待测样品中的植原体浓度极低时，比如葡萄组织样品中植原体的检测。据此提出了用于不同检测目的时的最佳引物。     

圆点印迹法检测葡萄斑点病毒技术研究

采用圆点印迹法检测GFkV，结果表明：Tris-HCl和PBS在提取GFkV时效果相同，在带毒葡萄植株中韧皮部的GFkV含量较高。     

Evaluation of two commercial PRRSV antibody ELISA kits with samples of known status and singleton reactors

Two commercial PRRSV ELISA kits (IDEXX and Bionote) were evaluated for their sensitivity and specificity using 476 PRRS-positive serum samples collected from 7 animal challenge experiments and 1,000 PRRS-negative sera. Both ELISA kits exhibited 100% sensitivity with sera collected 14 to 42 days post-infection, and the results from the kits were highly correlated (R²=0.9207). The specificity of IDEXX or Bionote kit was 99.9% or 99.7%, respectively. In addition, the Bionote ELISA kit was used to examine 100 sera that were determined to be falsely positive either by IDEXX 2XR or 3XR ELISA, and only 7 of these samples were found to be positive. These results indicate that both ELISA kits exhibited similar levels of sensitivity and specificity and would complement one another for the verification of false-positive samples.     

Feasibility analysis of NIR for detecting sweet corn seeds vigor

This paper explores the feasibility of particle-based detection and grading of seed vigor based on a self-built seed single-granulation device using near infrared spectroscopy (NIRS). Sweet corn with uniform kernel size was used for this study. The seed samples were divided into three types, they were normal seeds, artificially aged seeds and heat-damaged seeds. A 2-part spectral acquisition of each seed were performed, one for the collection of seeds that fall into the detection zone within the separation pipe, another was on the static platform, whose collection was performed on 5 faces of each seed. Partial least squares discriminant analysis (PLS-DA) was used to classify the original data of the seeds. In the 2 parts, the discriminant results of the unprocessed normal seeds and the artificial accelerated aging seeds, the untreated normal seeds and the heat-damaged seeds showed that classification accuracy was higher than 98%. The research indicates that the spectral data of different positions of seeds can reflect their activity information, and it is feasible to detect and classify seeds in real time in the detection area of the separation pipeline.     

粉条加工中添加石蜡的检测技术研究

【目的】探讨粉条加工过程中非法添加石蜡的测定方法,为食品监管提供保障。【方法】先以石蜡成品摸索石蜡测定的气相色谱质谱条件,再以石蜡成分C18H38和C24H50进行回收率和相对标准偏差试验。【结果】不同预处理方法中,超声萃取处理的C18H38和C24H50在粉条中的萃取回收率最高,分别为93.5%和91.0%;20min超声时间萃取粉条中烷烃成分较为适宜;所建立的粉条中石蜡含量测定方法的相对标准偏差为3.6～4.8,样品回收率均大于90.0%,达到定量检测要求。【结论】所建立的石蜡检测方法操作简单,测定结果准确可靠,可用于粉条中石蜡成分的定性及定量测定。     

产碳青霉烯酶肠杆菌科细菌检测方法研究进展

抗生素的不合理使用使得细菌耐药性快速产生及传播，细菌耐药性已然成为重大公共卫生问题。碳青霉烯类抗生素在临床治疗中表现良好，常作为治疗多重耐药菌感染的最后一道防线，碳青霉烯耐药菌的出现对公共卫生安全产生了极大的威胁。产碳青霉烯酶是碳青霉烯耐药菌耐药的主要机制，因此，对产碳青霉烯酶肠杆菌科细菌的检测至关重要。目前，对于碳青霉烯酶的检测手段以表型检测和分子生物学检测方法为主。表型检测一般易于操作，成本低廉，便于在临床进行，但在检测效率和检测用时方面差强人意。分子生物学及其他新型技术在保持高特异性和敏感性的基础上能做到快速、高通量检测，但试验检测的成本高，需专业设备和人员，部分技术在碳青霉烯酶检测领域仍有待开发。当前，多种碳青霉烯酶的检测方法可针对不同的需求和目的，并没有一项检测能满足所有情况。进行检测时，试验所需时间、操作难度、经济成本等都需列入考量，应根据实际情况选择合适的检测方法。笔者通过结合当前背景，归纳总结产碳青霉烯酶肠杆菌科细菌的检测方法，从试验特异性与敏感性、可操作性、检测时间、试验成本等方面分析比较各个方法特点，以期为之后新的检测方法的开发及现有方法的改良研究提供切入点，为临床监测与检测方案的制定和实行提供思路。     

检测马疱疹病毒4型抗体间接ELISA方法的建立及应用简

In order to establish an indirect ELISA method for detection of equine herpesviruses type 4 （EHV4） antibody, the glycoprotein G （gG） protein with specific epitope worked as a detection antigen. After optimizing conditions, the indirect ELISA method was developed successfully，specificity and repeatability tests were determined. The result was that the EHV4 gG only reacted with antibodies against EHV4;but not with antibodies against EHV1; both the intro-batch and inter-batch variation coefficiencies were lower than 10%.Concordance of the indirect ELISA relative to commercial EHV1/4 antibody kit was above 90%.The results indicated that the indirect ELISA method could be used for the detection and epidemiological surveys of EHV4 infection.