绿僵菌属血清学的初步研究

本文采用凝集试验和免疫电泳的方法对不同来源的七株绿僵菌属真菌进行了免疫学对比研究。抗原是孢子悬液和菌丝体清液,通过对角兔接种抗原而获得抗血清。每一种抗原与其同源抗血清和异源抗血清进行交叉试验,并对凝集试验的结果以及免疫电泳反应产生的沉淀弧数目和免疫电泳图谱加以对比分析,试验结果表明：不同菌株间存在明显的抗原类似性,同时各菌株间也表现出一定的抗原专一性。根据试验数据对供试菌株进行血清学分型的结果与形     

对流免疫电泳与其他的血清学试验在诊断人布氏病上的比较

<正> 在标准快速血清学试验的比较中使用了65个临床诊断为布氏病的病人的血清。孟加拉红试验与标准试管凝集试验所到结果的相关性很好,而快速平板凝集试验和Coombs (抗球蛋白)试验则较差。用特异的抗——人免疫球蛋白血清吸收病人的血清表明IgM在孟加拉红试验中是有作用的而在Coombs试验中则没有作用。IgG和IgA在这两个试和Co验中都起作用。在凝集试验、孟加拉红试验、Coombs试验中起着最重要的作用的A和M抗原之外,用沉淀试验还检出了布氏其他的抗原部份。在对流——免疫电泳中有一种蛋白质抗原与94％的血清起反应。根据两组血清所得的结果,孟加拉红试验和对流-免疫电泳试验对于诊断临床布氏病是最有希望的方法。由于使用布氏抗原不同试验的性质也不同。     

固氮螺菌的血清学研究——Ⅱ.固氮螺菌五个血清型的抗原定性分析

本文采用琼脂双扩散、对流电泳及免疫电泳等技术对固氮螺菌五个血清型的代表菌株进行抗原定性分析。试验结果说明:五个不同的血清型的可溶性抗原具有共同的抗原成份,但各自存在着特异性抗原。     

用免疫学方法检测脑脊液中抗原诊断隐球菌脑膜炎

1．本文报道成功地用改良的对流免疫电泳及微量补体结合技术检测隐球菌脑膜炎病人脑脊液中多糖体抗原的方法。对流免疫电泳的改进要点是电泳凝胶板采用国产琼脂糖。由于琼脂糖电渗流很弱,将凝胶板放在冰箱中7天后使用或在紧急待用时,在琼脂糖中掺入30％普通琼脂就可以使抗体产生足够的负向移动。改良对流免疫电泳后敏感度提商10倍左右。 2．本室制成的1号抗血清与本市所收集的9个菌株都能起凝集反应,也能与9个菌株的多糖体抗原起免疫反应,特异性和敏感性都是比较满意的。 3．用多糖体对1号抗血清的吸收试验,初步证叫所有多糖体实际上都可移走1号扰血清的凝集活性,但是用5号菌株吸收后,残留有少量的对几个菌株的凝集素,这可能是指它们存在有不能完全吸收的微量抗原。 4．用这批抗血清检测了临床几年来收集的16份脑脊液标本和11份血清标本,其中三例临床确诊为新型隐球曲脑膜炎,我们的抗原测定也全部阳性,其它疾病的24份标本抗原测定 全部阴性。 5．测定抗原的效价能反映出病情严重程度,迁延和治疗效果。     

百日咳的血清学诊断

<正> 血清学试验可确诊百日咳,用凝集试验或补体结合试验(CFT)方法能证明百日咳抗体。凝集试验检出不耐热血清型抗原的1,2,3型和耐热的菌体抗原的抗体。活的或福尔马林杀菌的菌悬液检出不耐热抗原的抗体。新近的两个试验中应用了不同的CFT抗原。1972年Bradstreet等用60℃加热1小时菌悬液的远沉上清作抗原,菌株血清型抗原并不影响血清效价,提示发现的主要抗体是对抗原1的(所有菌株所共有)而不是型特异抗体。苏格兰联合研究用新近分离的1,3型菌株并用NaOH37℃2小时提取抗原,他们没有证明是否测出了型特异抗体。 用间接血凝试验(IHA)测定人血清百日咳抗体的报导很多,除了Schuber等用百日咳可疑病例的双份血清做过试验,没有任何别的作者指出抗原附着到红血球上。     

对流免疫电泳对El Tor型霍乱弧菌与不凝集弧菌抗原相关性的初步探讨

本文对37株NAG 8株El Tor弧菌的抗原性进行了初步分析,用对流免疫电泳不同程度地出现有交叉沉淀线,这说明它们之间有血清学相关性;通过血清吸收试验,发现“H”抗原吸收后的血清大部分仍出现沉淀线,而用同一菌株的可溶性抗原吸收后的血清与5种NAG可溶性抗原进行对流免疫电泳,除少数仍有沉淀线外,大部分均消失,因而证明它们之间的抗原有相关性,而此相关部分并非弧菌共同的H抗原,以对流电泳与糖发酵反应双盲试验结果也证明二者之间亦有一定联系。本文还介绍了用去氧胆酸钠裂解细菌的方法、原理和注意事项。     

2308、M28、S1330、16M四株布鲁氏菌灭活参数研究

【目的】比较不同灭活方法对布鲁氏菌灭活的效果,确定灭活参数,为制备布鲁氏菌灭活抗原提供参考。【方法】将4株布鲁氏菌参考强毒株2308(牛种)、M28(羊种)、S1330(猪种)以及16M(羊种)分别经大豆酶消化蛋白胨(TSA)培养基培养繁殖后,用生理盐水制成(4-8)×1010 CFU/m L的菌悬液,分成等份于80 oC灭活不同时间,另将同样浓度的菌悬液分别用不同浓度甲醛于37 oC灭活不同时间,通过灭活检验,确定灭活效果。取经甲醛和热灭活的16M抗原,分别以1×1010 CFU/只剂量皮下注射1.5-2.0 kg家兔2只,免疫6周内,每周采血用虎红平板凝集试验(RBT)和试管凝集试验(SAT)测定抗体效价。【结果】80 oC、5 min可灭活2308、S1330和16M三种菌株,80 oC、10 min可灭活全部4种菌株。0.2%甲醛灭活7 d,4种试验菌株均不能被彻底灭活;0.4%甲醛在12 h内只能灭活16M,72 h可灭活M28;0.4%甲醛灭活2308和S1330两次试验结果差异较大。0.6%甲醛可在72 h内灭活4种试验菌。不同方法灭活的16M抗原免疫家兔后,其血清抗体虎红平板凝集和试管凝集效价消长趋势基本一致,甲醛灭活的抗原免疫原性略高于热灭活抗原。【结论】80 oC热灭活和0.6%甲醛灭活均可用于对布鲁氏菌的灭活,且不影响布鲁氏菌的免疫原性。     

茶毛虫核型多角体病毒的血清学特性

用免疫双扩散、对流免疫电泳、酶联免疫吸附试验(ELISA),固相免疫电镜(SPIEM)等技术,对茶毛虫核型多角体病毒(EpNPV)的抗原特性及与其它10种核型多角体病毒的血清学关系进行分析。结果表明,EpNPV粒子的抗血清只能与EpNPV粒子起反应,不与EpNPV的多角体蛋白及其它10种昆虫核型多角体病毒(NPV)粒子发生交叉反应;EpNPV多角体蛋白抗血清除了和其同源的多角体蛋白起反应外,还能和其它两种NPV的多角体蛋白起反应。以上结果说明了EpNPV的结构蛋白具有较高的抗原特异性,而多角体蛋白则没有种间特异性。同时将固相免疫电镜技术应用到昆虫病毒的血清学检测中,取得了较为理想的结果。     

亚硝酸诱变导致枯草杆菌中L-丙氨酸脱氢酶对L-谷氨酸脱氢酶的转换——Ⅲ.野生型中有与GDH抗原性上类似的蛋白质的存在

由枯草杆菌野生型菌株的ADH制剂所制备的抗ADH抗血清能中和ADH与GDH。免疫电泳试验的结果指出抗ADH抗血清中含有二个抗体;一为抗ADH抗体,另一为抗GDH抗体。在野生型细菌的无细胞抽出液中亦找到了与GDⅡ在抗原性上近似的蛋白质即GDH-ORM。5株不同来源的野生型细菌,细胞内仅有ADH而没有GDH活力,但均有GDH-CRM的存在。葡萄糖能促进GDH~+型变种的GDH的合成,同时也促进野生型细菌的GDH-CRM的合成。但是葡萄糖对GDH合成的促进作用要比其对GDH-ORM的作用显著得多。作者对野生型细菌的CRM与变种中GDH的形成的遗传学关系进行了讨论。张景六同志参加技术工作;在制备抗血清过程中,承上海生物制品研究所协助,谨此致谢。     

小肠结肠炎耶氏菌Vi抗原的研究

从小肠结肠炎耶氏菌发现ⅵ抗原,这是首次报道。用ⅵ抗原的抗血清测定菌株的毒力,与vw抗原和毒力因子血清测定的结果完全一致,即仅毒力株有ⅵ抗原,无毒株则无。故可肯定ⅵ抗原是小肠结肠炎耶氏菌的一种毒力抗原。利用ⅵ抗血清测定菌株的毒力,是非常简便的。     

Release of Vi antigens from Salmonella typhi: implications for virulence and diagnosis

The release of Vi antigens from three clinical isolates of Salmonella typhi was measured by a Vi-specific monoclonal antibody. Large quantities of Vi antigens were detected in the culture supernates from all three strains using either passive latex agglutination or rocket immunoelectrophoresis. Vi antigens were also detected in broth cultures of S. typhi containing about 10(5) cells/ml using the sandwich enzyme linked immunosorbent assay. The significance of this finding in relationship to the virulence and the diagnosis of S. typhi was discussed.     

Release of Vi antigens from Salmonella typhi: implications for virulence and diagnosis

Abstract The release of Vi antigens from three clinical isolates of Salmonella typhi was measured by a Vi-specific monoclonal antibody. Large quantities of Vi antigens were detected in the culture supernates from all three strains using either passive latex agglutination or rocket immunoelectrophoresis. Vi antigens were also detected in broth cultures of S. typhi containing about 10⁵ cells/ml using the sandwich enzyme linked immunosorbent assay. The significance of this finding in relationship to the virulence and the diagnosis of S. typhi was discussed.     

Latex agglutination test for diagnosing pneumococcal pneumonia in children in developing countries.

OBJECTIVE--To prepare and assess the sensitivity and specificity of a latex agglutination test specific for the serotype of antigen in diagnosing pneumococcal pneumonia in Gambian children. DESIGN--Comparison of agglutination test specific for serotype with culture of blood and lung aspirates, countercurrent immunoelectrophoresis, and commercial latex agglutination tests in diagnosing pneumococcal pneumonia. Cross reaction studies and investigation of 102 control children to determine specificity of agglutination test specific for serotype. SETTING--General medical ward of Medical Research Council laboratories, The Gambia. PATIENTS--101 Gambian children aged between 2 months and 10 years admitted with severe pneumonia. INTERVENTIONS--Serum samples were boiled and treated with edetic acid, and urine samples were boiled and concentrated 25 times before testing. END POINT--A latex agglutination test specific for the serotype of pneumococcal antigen that is sensitive and highly specific for detecting pneumococcus in the urine of patients with pneumococcal pneumonia. MEASUREMENTS AND MAIN RESULTS--Concentrated urine samples from 16 of the 21 children (76%) with pneumococcal pneumonia established by results of culture of blood or lung aspirates gave a positive result with the agglutination test specific for serotype, whereas only four of the 102 urine samples obtained from control children without pneumonia gave positive results. The serotypes of antigens detected in the urine of children with pneumococcal pneumonia and the serotypes of pneumococci isolated from cultures of blood or lung aspirates were the same in all cases. CONCLUSIONS--When performed on urine samples the agglutination test specific for serotype has a high specificity and is more sensitive than culture of blood or lung aspirates, commercial agglutination tests, or countercurrent immunoelectrophoresis in identifying pneumococcal pneumonia. It is easy to use and should be especially useful in communities with limited laboratory facilities.     

Membrane composition and virus susceptibility of Acholeplasma laidlawii.

The membrane composition of 11 strains of Acholeplasma laidlawii, including three strains persistently infected with mycoplasmaviruses MVL51, MVL2, and MVL3, was studied and correlated with mycoplasmavirus sensitivity. Membranes of the strains had similiar sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns, and all strains were inhibited by an antiserum produced against membranes from one of the strains. The amounts of integral membrane proteins solubilized by the nonionic detergent Tween 20 differed considerably. Therefore, characteristic crossed immunoelectrophoresis patterns were obtained for each strain. Strains persistently infected with MVL2 and MVL3 were notably different from the noninfected host. The ability to propagate any of the viruses was not correlated with sodium dodecyl sulfate-polyacrylamide gel electrophoresis or crossed immunoelectrophoresis patterns. The persistently infected strains had a characteristic lipid composition. MVL51-resistant strains, including a resistant clone selected from a sensitive strain, were characterized by a large monoglucosyldiglyceride/diglucosyldiglyceride ratio and trace amounts of diphosphatidylglyceol (as opposed to the sensitive strains). Differences in lipid composition in A. laidlawii seem to affect the relationship between cells and viruses.     

Cell-free Pseudomonas vaccine. III. Immunochemical analysis of purified Pseudomonas aeruginosa protein antigens and protective properties of antisera against these antigens

The immunochemical analysis of isolated and purified antigens A and B obtained from P. aeruginosa, strains 868 (serogroup O3 according to Lányi or immunotype 3/7 according to Fisher) and 170015 (serogroup O7 or immunotype 2), was carried out. Rabbit antisera to proteins A and B, as well as to the initial aqueous extracts and partially purified aqueous extracts, were obtained. Cross activity between the protein antigens of different strains was established by the methods of immunodiffusion and two-dimensional immunoelectrophoresis. Isolated proteins A and B contained both common and specific antigenic determinants detected by the method of two-dimensional immunoelectrophoresis. The immunization of rabbits with proteins A and B was found to stimulate the synthesis of protective, probably species-specific, antibodies.     

Antigens common to human malignant tumors and certain species of microorganisms

Agglutination, gel precipitation and immunoelectrophoresis demonstrated the presence of antigens common for human malignant tumors of different sites and BCG and Listeria monocytogenic microorganisms. The radioimmunoassay of sera in the agglutination test showed that these sera reacted with tumor cells rather than with cells from normal human tissues. The common antigen had the electrophorectic mobility in the beta-globulin zone.     

Comparison of Purified Alpha-Toxins from Various Strains of Staphylococcus aureus

Alpha-toxin from five strains of Staphylococcus aureus, including Wood 46, was purified by isoelectric focusing. The alpha-toxins obtained from different strains were similar. The isoelectric point of the purified toxins was 8.65 +/- 0.15. Sharp concentration peaks were not always obtained. In the ultracentrifuge the alpha-toxins migrated usually as three peaks which could be dissociated with propionic acid to yield one peak. A single line of identity was obtained in immunoelectrophoresis when a heterologous antiserum was reacted with the five purified toxins. It was concluded that the widespread use of the Wood 46 strain for the production of alpha-toxin is justified.     

Taxonomic and serologic studies on Micropolyspora faeni and Micropolyspora strains from soil bearing the specific epithet rectivirgula.

The results of serological studies on six strains of Micropolyspora faeni from hay, sputum and plant debris, and five strains of Mip. rectivirgula from soil indicated no significant differences between the two species. Antisera raised in rabbits against purified antigens of the type strains were used to compare the 11 strains by immunoelectrophoresis. The detailed antigenic composition of the type strains was also determined by two-dimensional immunoelectrophoresis against specific rabbit antisera and pooled serum samples from patients suffering from farmer's lung. Cross-reacting antigens were identified by intermediate gel immunoelectrophoresis. The close similarity of the two species was confirmed by the results of 60 morphological physiological and biochemical tests applied to the 11 strains. We consider that the strains belong to a single species and propose that the specific epithet faeni be conserved for the taxon.     

Trypanosoma cruzi: surface antigenic determinants

A fraction (FAd) capable of inhibiting specific agglutination reactions of anti-epimastigote sera (anti-LE) was obtained by extracting the sediment of lyophilized epimastigote lysates (LE) with 0.05 M phosphate buffered saline, at 37 degrees C for 1 h. These conditions favored the action of parasite proteinase whose presence was detected by tandem-crossed immunoelectrophoresis experiments. As expected from the proteinase properties, the addition of 2-mercaptoethanol or sodium iodoacetate to the extracting solution resulted, respectively, in either increased or decreased amounts of protein in the resulting FAd. FAd components could be precipitated by the addition of Concanavalin A, methylated albumins or 0.1 N HCl. This fraction presented a single component when subjected to electrophoresis in 1% agarose gel with an electrophoretic mobility 1.2 times higher than that of human albumin. FAd component(s) were unable to penetrate 15% polycrylamide gel matrix unless 1% SDS was used. Under this condition four glycopeptide components, with Rm of 0.5, 0.55, 0.6 and 0.86, were detected. The antigenic determinants present in FAd resisted heating at 100 degrees C for 30 min and the prolonged action of pronase. However, these determinants were completely destroyed by the action of 25 mM sodium periodate, thus suggesting polysaccharide characteristics. Immunization of rabbits with FAd induced the production of antibodies that were unable to precipitate with either FAd or with parasite proteinase. These antibodies exhibited positive agglutination reactions with epimastigote forms and positive immunofluorescence and immunoperoxidase reactions with trypomastigote and amastigote forms of the different strains tested. FAd was able to inhibit these reactions as well as those obtained with anti-LE and anti-FA immune sera, whereas purified proteinase was unable to inhibit any of these reactions.     

Detection of Hepatitis Associated Antigen by the Latex Agglutination Test

Hepatitis associated antigen may be detected quickly and reliably by the latex agglutination test, using antiserum from guinea pigs immunized with the antigen. The latex test has a sensitivity comparable to the counter current immunoelectrophoresis technique.