Estimation and Identification of Streptococcus dysgalactiae Protective Antibody in Sera of Rabbits and Cattle

Rabbit and cow anti-Streptococcus dysgalactiae sera were tested by bacterial agglutination, complement fixation, hemagglutination, and immunodiffusion for the presence of antibody. The results of these tests were compared with mouse-protection studies on the same serum to estimate which in vitro test would best reflect the in vivo protective capacity of serum. Identification of the antibody constituents responsible for the mouse protection, hemagglutination, and complement fixation titers were established by reacting whole and diluted antisera with mercaptoethanol before and after testing. Results indicate that the complement fixation test may be a more accurate indicator of IgG protective bovine and rabbit antibody, whereas the hemagglutination test may more readily reflect a wider range of protective antibody levels and IgM. The complement fixation test showed some shared responses to IgG and IgM in both the rabbit and cow, whereas the IgM components seemed to be the predominant factor influencing hemagglutination titers in the rabbit and more so in the bovine. Mouse protection tests with mercaptoethanol-treated cow and rabbit sera indicate that the protective capacity of these antisera is shared between IgM and IgG components.     

Comparative diagnostic value of methods for detecting dysentery antigens in substrates of the patient's body

The comparative evaluation of different immunological methods, such as the enzyme immunoassay, the aggregate hemagglutination test and the complement fixation test, used for the detection of specific Shigella antigens in biological body substrates obtained from 287 patients with acute dysentery caused by S. sonnei, S. flexneri and S. newcastle has been carried out. The enzyme immunoassay and the aggregate hemagglutination test most effective (97.5 +/- 0.5 and 92.4 +/- 0.9, respectively), the object of study being the patients' blood taken at the early stages of the disease. The diagnostic specificity of these methods has proved to be 98.7 +/- 6.7 and 95.2 +/- 1.4, respectively.     

Comparison of Methods for Coccidioidomycosis Complement Fixation

A Laboratory Branch Task Force of the National Communicable Disease Center has proposed a standardized complement fixation procedure (LBCF) and an adaptation of this to microtitration techniques (MT) as uniform methods for performing complement fixation (CF) tests. A common procedure should make CF results from one laboratory more comparable to another. In addition, it would be preferable if the common procedure reproduced the titer levels of a testing procedure which is to be replaced, particularly when valid clinical interpretations have been derived from the latter. Replicated sets of sera were tested by the LBCF, MT, and the standard Smith CF procedure for coccidioidomycosis. Results with all three procedures were highly reproducible within an acceptable one-tube variation of a twofold dilution series, but the frequency of one-tube variations was greater with the MT method than with the other two. There was no statistical difference in the titers obtained with the Smith and LBCF procedures, but there was a significant difference when the MT results were compared to those with the Smith method. The LBCF method should be acceptable as a standardized and uniform CF procedure for coccidioidomycosis, subject to comparative testing between different laboratories.     

A Simple Method for Histochemical Demonstration of Viral Inclusion Bodies in Cell Monolayers in Microtitration Plates

The present paper describes a Bouin's-formalin fixation and Giemsa staining procedure for demonstrating viral cytoplasmic inclusion bodies in cellular monolayers in microtitration plates. Monolayers are fixed in Bouin's fixative for 15 min followed by 10% neutral buffered formalin fixation overnight. After fixation, the monolayers are stained with 4% (v/v) Giemsa stain. The method is superior to the separate use of formalin, methanol or Bouin's fixation-staining methods and compares favorably to immunocytochemical techniques for sensitivity.     

A simple method for histochemical demonstration of viral inclusion bodies in cell monolayers in microtitration plates

The present paper describes a Bouin's-formalin fixation and Giemsa staining procedure for demonstrating viral cytoplasmic inclusion bodies in cellular monolayers in microtitration plates. Monolayers are fixed in Bouin's fixative for 15 min followed by 10% neutral buffered formalin fixation overnight. After fixation, the monolayers are stained with 4% (v/v) Giemsa stain. The method is superior to the separate use of formalin, methanol or Bouin's fixation-staining methods and compares favorably to immunocytochemical techniques for sensitivity.     

Trial of an erythrocyte antigenic diagnostic agent for detecting antibodies to Coxiella burnetii

A new method for the preparation of Q-fever erythrocyte antigenic diagnosticum, adaptable to large-scale production, has been developed, and the diagnosticum thus obtained has been found to be highly specific and sensitive. The combined use of the complement fixation and passive hemagglutination tests enhances the effectiveness of the serological study, as it not only ensures a more complete detection of antibodies to C. burnetii, phase I, in humans and cattle, but also gives more precise indications on the time of infection.     

Use of the indirect hemagglutination test for the study of ornithosis. Report 2. Preparation and approbation of the dry ornithosis erythrocyte diagnostic agent

An original preparation--dry ornithosis erythrocytic diagnostic agent for the indirect hemagglutination test was prepared on the basis of formalinized tannin-treated sheep red blood cells and group-specific phospholipid antigen of the causative agent of ornithosis. This diagnostic agent retained its specific activity for 18 months (observation period). The use of this preparation considerably facilitated the method of performance of this test, this offering a possibility of its wide application in practice. A sufficiently high sensitivity and specificity of the indirect hemagglutination test with the suggested diagnostic agent, in comparison with the complement fixation test was demonstrated.     

Preparation and properties of antisera to glycolipid of guinea pig erythrocyte membrane

Antibodies against a glycolipid of guinea pig erythrocyte membranes were prepared in rabbits by immunization with guinea pig erythrocyte stroma or the purified glycolipid, gangliotriaosylceramide. The antibodies agglutinated guinea pig erythrocytes. The specificity of antibodies could be revealed by several immunochemical methods, including inhibition of hemagglutination, immunodiffusion, agglutination of liposomes, and complement fixation. The antibodies were specific for gangliotriaosylceramide.     

Immunological study of the occurrence of staphylococci with skin antigens in various experimental animals]

Experiments were conducted on rabbits; primary and secondary administration of staphylococcus vaccine was regularly accompanied by the production of antibodies not only to a staphylococcus antigen, but also of antibodies reacting with an extract of homologous kidneys, myocardium and the skin. The presence in the pathogenic staphylococcus of an antigen affiliated to proteins of the skin and kidneys of rabbits and mice was shown by the method of cross sorption of antistaphylococcus and antiskin sera by a suspension of the staphylococcus or skin antigen with the use of the complement fixation test. Indirect hemagglutination and immunofluorescence. Such antigen was absent in nonpathogenic bacteria isolated from the skin extracts.     

微量法与试管比色法检测ALT的比较

经实验建立了一种用于检测血浆中丙氨酸氨基转移酶(ALT)含量的微量测定法,并与比色法（传统赖氏法)进行了比较。用两种方法检测定值血清、室内质控及样品并比较标准曲线后,结果无显著性差异,同时微量法重复性较好,结果表明微量法测定ALT酶活力可以替代比色法测定血浆中ALT含量,适合大批量血浆ALT含量的快速检测。     

Staphylococcal antigens cross-reacting with mammalian tissues and their role in inducing immunopathological processes

In the reaction of cross adsorption of immune sera to white mouse skin and staphylococcus with the corresponding antigens, the presence of an antigen related to white mouse skin and kidney antigens has been established in Staphylococcus aureus strain Wood-46 (not producing protein A) by means of the complement fixation test and the passive hemagglutination test. The capacity of staphylococcal antibodies and their fluorochrome-labeled fragments to specifically stain the cells of the epidermis and the renal tubules of mammals has been demonstrated. Staphylococcus epidermidis has proved to be unrelated to mammalian tissues. The significance of the data obtained in this investigation for the practice of the transplantation of organs and tissues, for more accurate determination of the pathogenesis of staphylococcal infections and for the development of immunologically safe vaccines in discussed.     

Comparative evaluation of the commercial Sevatest ELISA immunoenzyme set and other serological tests for detecting Toxoplasma gondii antibodies

This work analyzes the results of 4 serologic tests used for the diagnosis of toxoplasmosis: the complement fixation (CFT), indirect immunofluorescence (IIF), passive hemagglutination (PHAT) tests, and the enzyme-linked immunosorbent assay (ELISA). The last-mentioned one was made with the use of the commercial kits Sevatest ELISA IgG/Toxo Micro I. The results of ELISA were in good correlation with those yielded by the traditional tests: 70% coincidence with CFT, 80% with IIF, 84% with PHAT; besides, ELISA has shown a higher sensitivity in the screening of sera.     

Automated Microtitration Test for Antistreptolysin O

A commercially available instrument that automatically makes serial dilutions and delivers reagents was used for the determination of antistreptolysin O titers in serum. The automated method was compared with tube-dilution and manual microtitration techniques. It gave higher reproducibility of results and was quicker to perform than both the other tests; and it was much more economical in reagents than the tube test. The automated technique is considered to be the best of the three methods when more than a small number of specimens are examined at one time. It is now in routine use in our laboratory.     

Use of the indirect hemagglutination reaction in studying ornithosis infection. III. The diagnostic value of the IHR with an ornithosis erythocytic diagnosticum]

In both experimental and clinical conditions the passive hemagglutination test (PHAT with the use of an ornithosis erythrocytic diagnostic preparation was found to be sufficiently sensitive and specific as compared with the complement fixation test (CFT), a routine testing method. The study of the dynamics of immune response in infected animals and ornithosis patients allowed to regard the PHAT as a comparatively early method of serological analysis. Hemagglutinins were also found to circulate in the patients' blood sera only for a short time (on the average for 1 1/2--2 months). The CFT and the PHAT with erythrocytic diagnostic preparation, used in combination, will make it possible not only to diagnose ornithosis in patient more effectively, but also to differentiate between the cases of infection and anamnestic reaction.20     

Comparison of the Reliability and Sensitivity of Three Serological Procedures in Detecting Antibody to Yersinia pestis (Pasteurella pestis)

Three serological procedures, the agar-gel precipitin inhibition, the complement fixation, and the indirect hemagglutination tests, were used to detect and measure antibody to Yersinia pestis in the sera from 383 individuals. Although all three tests were useful in detecting plague antibody, the most reliable and sensitive test procedure was indirect hemagglutination.     

Immunoenzyme analysis in a system of serologic evaluation of louse-borne typhus vaccines

The blood sera of persons immunized with different typhus vaccines have been studied in the complement fixation test, the indirect hemagglutination test and the enzyme immunoassay. The data thus obtained indicate that the enzyme immunoassay is highly sensitive and can be universally used for the determination of antibodies to Rickettsia prowazekii after primary and booster immunization with different typhus vaccines. This method detects specific antibodies both at an early period and, which is of particular importance, at a remote period after immunization (3 years later) when complement-binding and hemagglutinating antibodies are absent. This is seemingly indicative of the two-phase character of postvaccinal immunity induced by live typhus vaccine.     

Use of the passive hemagglutination test to diagnose toxoplasmosis]

The authors describe a method of obtaining toxoplasma erythrocytic diagnostic agent by sensitization of formalinized tannin-treated SRBC with purified toxoplasma antigen isolated by fractionation of complete toxoplasma antigen on Sephadex G-100. Comparative experiments with titration of sera of persons with suspected toxoplasmosis were conducted; the passive hemagglutination test with the antigen obtained proved to be highly sensitive in comparison with immunofluorescence and complement fixation tests.     

Current state of serological tests used to detect blood parasite infections

Various serologic tests used for immunodiagnosis of blood parasite infections are discussed, and the advantages and limitations of each are noted. The procedures include complement fixation, indirect fluorescent antibody, indirect hemagglutination, agglutination, and soluble antigen fluorescent antibody tests. The influence of the quality of antigen on the reliability of the tests is stressed, and the fact that no single procedure is suitable for all applications is emphasized. Problems peculiar to preparation of antigens from intraerythrocytic parasites are noted. In addition, an adaptation of the quantitatively standardized complement fixation procedure for critical, objective evaluation of antigens is described.     

Simple method for quantitation of cell-bound protein A on Staphylococcus aureus cells by means of hemagglutination with sheep erythrocytes differentially sensitized with rabbit antibody and its clinical application

A simple method for quantitation of cell-bound protein A (SpA) on organisms of Staphylococcus aureus was successfully devised by using hemagglutination between staphylococcal organisms and a series of sheep erythrocyte suspensions sensitized with different amounts of anti sheep erythrocyte rabbit antiserum. The validity of the principle and the reproducibility of the method presented here were precisely analyzed and the details of the method are presented. The hemagglutination was quantitatively inhibited both by normal rabbit serum and by soluble SpA. Using the method presented here, 376 strains of S. aureus freshly isolated from clinical materials were subjected to SpA quantitation in cell-bound form. According to the results, there was an unexpected distribution profile in the amounts of cell-bound SpA among the clinical isolates, which showed a two-peak pattern. A possible usefulness of the method presented here in clinical investigations is briefly discussed also.     

Echinostoma lindoense: larval antigens from the snail intermediate host, Biomphalaria glabrata

Passive hemagglutination using chromic chloride proved to be a rapid and useful method for a study of minute quantities of antigen extracted from larval Echinostoma lindoense (Sandground and Bonne), a trematode that develops in the snail intermediate host, Biomphalaria glabrata (Say). Parasite rediae were initially fragmented by three different procedures. Their soluble proteins were separated into two bands by electrophoresis on cellulose acetate, and into three fractions by molecular sieve chromatography. Rabbit antiserum was prepared from six weekly intramuscular injections of soluble redial protein in complete Freund's adjuvant, followed after 1 month by a single inoculation of alum-precipitated antigen. Antiserum was absorbed free of anti-snail antibodies and the immune complexes were removed by ion-exchange chromatography over DEAE-cellulose, producing an immunochemically pure IgG. Study of the rabbit anti-trematode antibody by precipitation, complement fixation, hemagglutination (HA), and inhibition of HA revealed a specific and high titered anti-larval antibody. These methods offer an approach to the problem of measuring the snail host's protective response against trematode reinfection; they also can be used to study the antigenic maturation of successive larval stages in the intermediate host.