采用改进的基质固相分散提取技术提取辣椒制品中的罗丹明B,对提取工艺进行优化.选择分散剂种类、分散剂与样品质量比、洗脱剂组成及洗脱剂用量作为研究因素,以提取液中罗丹明B的含量为考察指标,运用L16(45)正交试验优选出了辣椒面中提取罗丹明B的最优工艺条件:分散剂硅胶,分散剂与样品质量比例1∶1;洗脱剂为质量分数80％乙腈水溶液,洗脱剂用量20 mL.辣椒油中提取罗丹明B最优工艺条件是:分散剂中性氧化铝,分散剂与样品质量比例2∶1;洗脱剂为体积分数80％甲醇水溶液,洗脱剂用量20 mL.辣椒酱中提取罗丹明B最优工艺条件是:分散剂中性氧化铝,分散剂与样品质量比例3∶1;洗脱剂为质量分数80％乙腈水溶液,洗脱剂用量5 mL.该提取方法萃取工艺简单,操作时间短,所需有机溶剂种类少,属环境友好型萃取方式.

To develop a specific, rapid and convenient method based on molecular motor biosensor to detect food-borne rotavirus. A specific probe was encompassed the conservative region of rotavirus's VP7 segment, and a molecular motor detect device was constructed by connecting probes to F0F1-ATPase molecular motor through biotin-streptavidin system. This biosensor's sensitivity was 0.005 ng/mL for rotavirus RNA. Extracted virus RNA was conjugated with the biosensor separately, at the same time ATP was synthesized. By comparing fluorescence intensity, we can detect rotavirus RNA in samples. This method possessed specificity for rotavirus, without any cross-reaction with Hepatitis A virus and noroviris, and it could be accomplished within 1 h. We detected 15 samples using this method and the results were compared with RT-PCR results. This method is sensitive and specific for rotavirus, and it can be used to detect food-borne rotavirus.

[Objective] To establish a simple and rapid molecular typing method for Vibrio parahaemolyticus carrying and non-carrying virulence genes based on molecular motor.[Methods] Four probes specific to virulence genes tdh and trh,species-specific genes tlh and toxR of V.parahaemolyticus were synthesized,and four molecular motor biosensors were constructed by connecting probes to F0F1-ATPase molecular motors through biotin-streptavidin system,respectively.Ten strains of V.parahaemolyticus were classified by the biosensors,and the results were compared with PCR-Electrophoresis-Gel imaging results.Further more,the detection sensitivities and specificities of the molecular motor biosensors were studied.[Re-sults] There were ten strains carrying tdh and none carrying trh,while all ten strains carry tlh and toxR,which was consisitent with the results of PCR-Electrophoresis-Gel imaging.The detection limits of molecular motor biosensors for tlh,toxR,tdh and trh were estimated to be 1 pg/reaction system,and the detection limits of PCR-Electrophoresis-Gel imaging for tlh,toxR,tdh and trh were estimated to be 10 pg/reaction system.The molecular motor biosensors could recognize tlh,toxR,tdh and trh of V.parahaemolyticus specifically.[Conclusion] A molecular typing method was constructed based on molecular motor biosensors and was used to diagnose the pathogenicity of V.parahaemolyticus rapidly and specifically.The detection limits was 10 times higher than those of PCR-Electrophoresis-Gel imaging.The method is easy,rapid,time-saving and labor-saving,especially suitable for the basic laboratories of CDC and port quarantine departments to perform suiveillance and epidemiological traceability of cholera.

A rapid detecting method for Enterobacter sakazakii based on F0 F1-ATPase molecular motor biosensor was cons tructed. Specific ITS probe were connected with F0 F1-ATPase’s ε subunit by using avidin-biotin system, and then biosensors were combined with the test samples and negative sample, respectively. To compare the catalytic ATP synthesis, Enterobacter sakazakii DNA in the samples were tested. The result showed that the optimum conditions for detection were the concentration of chro ITS of 0. 019 mg/mL and Enterobacter sakazakii DNA concentration of 40 ng/mL. The result was consistent to that of the traditional detection methods and PCR detection method, showing this is in good agreement.

NaYF4 : Yb3+, Er3+, Tm3+ nanoparticles were prepared by microemulsion-hydrothermal method. Crystal phase, morphology and structure of the samples were characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The luminescence properties were studied by up-conversional fluorescence spectroscopy. The XRD patterns of as-prepared samples were in agreement with the PDF # 77-2042 of cubic NaYF4. SEM images of the particles showed that the samples were cotton-like spherical in shape and which were assembled by smaller nano-particles. The average size was 120 nm, while the shape was regular and the particle size was homogeneous. Under the excitation of 980 nm, the as-prepared particles could emit blue (438 and 486 nm), green (523 and 539 nm) and red (650 nm) light simultaneously. It can be seen from the color coordinates figure (CIE) that when doping concentration ratio of Tm3+ and E3+ increased from 0 to 2, the whole emitting light color of samples movedto green region. While the ratio was 1 : 1, pseudo white light was obtained. As the ratio changed from 2 to 7, the luminous color was moved to red region.

The core technology was that the F0F1-ATPase molecular motor biosensor on chromatophore was used to rapidly detect Vbrio cholerae.Specific ompW probe were connected with ε subunit of F0F1-ATPase by avidin-biotin system,and then biosensors were constructed,and the test samples and negative sample were respectively combined with biosensors.After the ATP synthesis was catalyzed by them for 30 min,the content of ATP and Vbrio cholerae DNA in the samples were measured.The amount of ATP synthesis could be measured via the measurement of the amount of H+,while the amount of H+ was normalized by the fluorescence intensity of F-DHPE.According to the results of our experiments,optimum conditions for detection was under the presence of 0.078 mg/mL chro ompW and 40 ng/mL Vbrio cholerae DNA.After detection of the actual samples,we found that the method was in good agreement with the traditional detection methods and PCR detection method.