2019级检验英语ppt课件

1、About immunology immune response 免疫反应免疫反应antigen-antibodymonoclonal antibody (McAb)单克隆抗体单克隆抗体polyclonal antibody多克隆抗体多克隆抗体recombinant antigen 重组抗原重组抗原serial dilutions系列稀释系列稀释high titer高滴度高滴度enzyme-linked immunosorbent assay (ELISA )酶联免疫吸附试验酶联免疫吸附试验horseradish peroxidase-conjugated goat anti-rabbit I

2、gG辣根过氧化物酶标记的羊抗兔辣根过氧化物酶标记的羊抗兔IgGAbout immunology precipitation : double diffusion test双扩散试验双扩散试验 gel immunoelectrophoresis凝胶免疫电泳凝胶免疫电泳agglutination: direct agglutination 直接凝集试验直接凝集试验 indirect agglutination间接凝集试验间接凝集试验 latex agglutination test; indirect hemagglutination testradioimmunoassay 放射免疫方法放射免疫

3、方法immunoblotting: PAGE 聚丙烯酰胺凝胶电泳聚丙烯酰胺凝胶电泳 polyacrylamide gel electrophoresisimmunofluorescence 免疫荧光免疫荧光 polymerase chain reaction (PCR) 聚合酶联反应聚合酶联反应restriction fragment-length polymorphism analysis (RFLP) 限制性片段多态性分限制性片段多态性分析析restriction endonuclease: BamH ; EcoR 限制性内切酶限制性内切酶 (5 -GAATTC-3 ) (3 -CTTAA

4、G-5 ) About molecular biologyWestern-blot 免疫印迹免疫印迹Southren-blot (DNA-DNA hybridization)Northern-blot (DNA-RNA hybridization)DNA probe DNA探针探针Immunochromatography 免疫层析免疫层析immunogold immunogold labelling rHIV-Ag1rHIV-Ag2anti-HIV McAbELISAELISA positive control 阳性对照阳性对照negative control 阴性对照阴性对照blank co

5、ntrol 空白对照空白对照coating overnight at 4C 包被过夜包被过夜blocked with 封闭封闭washed with 0.1% Tween 20 in PBS the secondary antibody 二抗二抗the absorbance 吸光度吸光度pregnancy 妊娠；受孕妊娠；受孕chorionic 绒毛膜的绒毛膜的gonadotropin 促性腺激素促性腺激素latex 乳汁；胶乳乳汁；胶乳polymer 多聚物多聚物chorioepithelioma 绒毛膜上皮癌绒毛膜上皮癌hydatid 包虫囊包虫囊stir 搅拌；动搅拌；动vigorous

6、ly 精力旺盛的；强壮的精力旺盛的；强壮的Lesson SevenPregnancy Testing PRINCIPLE: Human chorionic gonadotropin (HCG) appears in the urine of pregnant females, and determination of its presence generally indicates a positive test for pregnancy .Lesson SevenPregnancy Testing PRINCIPLE: Utilizing an immunologic technique

7、based on the principle of indirect agglutination, tests will detect HCG at levels of 1.5 to 2.5 International Units (IU) per milliliter. Since levels as low as 1.5 IU have been reported as early as 5 days after the first missed period, negative tests should be repeated within 1 to 2 weeks.Lesson Sev

8、enPregnancy Testing PRINCIPLE: Test components consist of an antigen in the form of suspension of HCG chemically bonded to latex polymer particles and an antiserum containing antibodies to HCG. Lesson SevenPregnancy Testing Lesson SevenPregnancy Testing PRINCIPLE: When the antiserum is mixed with ur

9、ine containing a detectable level of HCG, it is neutralized, and no agglutination occurs. This is a positive test for pregnancy. Urine that contains no HCG cannot neutralize the antiserum, resulting in agglutination of the latex reagent. This indicates a negative test for pregnancy.PRINCIPLE: Patien

10、ts with chorioepithelioma or hydatid mole(葡萄胎葡萄胎) frequently excrete HCG in the urine. Therefore a positive result obtained when testing these patients does not always indicate a positive test for pregnancy. Lesson SevenPregnancy Testing REAGENTS: Available as a kit from Roche Diagnostics .PROCEDURE

11、: 1. Drop 1 drop urine in circle on glass slide . 2. Add 1 drop antiserum reagent . 3. Stir and rotate gently for 30 sec . 4. Add 1 drop antigen reagent . 5. Mix over entire circle . 6. Rotate slide vigorously for no longer than 2 min while observing for agglutination . Lesson SevenPregnancy Testing

12、 INTERPRETATION: Report as 1. Positive test: No agglutination within 2 min. HCG present in the urine of pregnant women will inhibit agglutination of latex particles in the antigen reagent, resulting in a smooth, milky appearance after rotation.Lesson SevenPregnancy Testing INTERPRETATION: Report as

13、2. Negative test: Any degree of agglutination within 2 min. Lack of HCG in the urine of nonpregnant women permits agglutination of the antigen and antiserum reagents within a 2 min period, producing easily recognizable clumping(块块) of the latex particles. Lesson SevenPregnancy Testing To quantitate

14、Ag-specific IgG, serial dilutions of serum prepared from tail vein blood were added to ELISA plates coated with rSRS9 or rSAG1(50ng/well). After incubation(孵育孵育) with peroxidase-coupled goat anti-mouse IgG and its substrate(底物底物), and plates were read colorimetrically at 450 nm.Sandwich ELISA One of

15、 the most useful of the immunoassays is the two antibody sandwich ELISA. This assay is used to determine the antigen concentration in unknown samples. This ELISA is fast and accurate, and if a purified antigen standard is available, the assay can determine the absolute amount of antigen in an unknow

16、n sample. The sandwich ELISA requires two antibodies that bind to epitopes(表位表位) that do not overlap on the antigen. Sandwich ELISA This can be accomplished with either two monoclonal antibodies that recognize discrete (不连续的不连续的) sites or one batch of affinity-purified polyclonal antibodies. To util

17、ize this assay, one capture antibody is purified and bound to a solid phase typically attached to the bottom of a plate well. Antigen is then added and allowed to complex with the bound antibody. Unbound products are then removed with a wash, and a labeled second detection antibody is allowed to bin

18、d to the antigen, thus completing the sandwich. Sandwich ELISAThe assay is then quantitated by measuring the amount of labeled second antibody bound to the matrix (矩阵矩阵), through the use of a colorimetric substrate. Major advantages of this technique are that the antigen does not need to be purified prior to use, and that these