Complement Fixation Tests For Syphilis Research Articles

Determination of the Optimal Ratio of Cholesterol to Tissue-Extract Antigen in the Complement-Fixation Test for Syphilis

Summary and Conclusions There is an optimal ratio cholesterol:tissue-extract that exhibits maximal antigenic activity in the complement-fixation test for syphilis. With the antigens studied, increase in the ratio, cholesterol:extract, did not result in anticomplementary properties. To determine the optimal ratio, extract must be tested in such concentrations that the optimal amount of cholesterol will fall within the range of quantities soluble in alcoholic tissue-extract. Varying the concentration of alcoholic extract in alcohol does not alter the optimal ratio. Different concentrations of extract require proportionately different quantities of cholesterol for their maximal sensitization. Varying concentrations of the alcoholic extract in alcohol exhibit the same antigenic activity, provided the optimal quantities of cholesterol are added. Differences in concentration are compensated for in tests by changes in the saline dilutions yielding maximal reactions of given quantities of serum. As a result of these findings, the adjustment of cholesterol and tissue-extract for use as antigen in the complement-fixation test for syphilis may be based upon that optimal ratio of the two which exhibits maximal antigenic activity. The experimental procedures given offer a practical method for determining this ratio.

A NEW AND ECONOMICAL DESICCATING PROCESS PARTICULARLY SUITABLE FOR THE PREPARATION OF CONCENTRATED PLASMA OR SERUM FOR INTRAVENOUS USE: THE ADTEVAC PROCESS

Article1 August 1940A NEW AND ECONOMICAL DESICCATING PROCESS PARTICULARLY SUITABLE FOR THE PREPARATION OF CONCENTRATED PLASMA OR SERUM FOR INTRAVENOUS USE: THE ADTEVAC PROCESSJOSEPH M. HILL, M.D., DAVID C. PFEIFFER, M.E.JOSEPH M. HILL, M.D.Search for more papers by this author, DAVID C. PFEIFFER, M.E.Search for more papers by this authorAuthor, Article, and Disclosure Informationhttps://doi.org/10.7326/0003-4819-14-2-201 SectionsAboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinkedInRedditEmail ExcerptA practical and inexpensive desiccation process is vital to the development of the widespread use of plasma and serum (ordinary or convalescent) because it solves two main problems in this field. First, storage difficulties practically cease to exist. While it is true that liquid serum may be stored, even without refrigeration as emphasized recently by Levinson,1there is no margin of safety. Contamination with but one bacterium could quickly spoil this excellent culture medium if stored without refrigeration. Also at room temperatures, the antibodies, prothrombin and other valuable biological properties rapidly deteriorate. By contrast, serum stored in the dry so-called...Bibliography1. LEVINSONNEUWELTNECHELESE SOFH: Human serum as a blood substitute in the treatment of hemorrhage and shock, Jr. Am. Med. Assoc., 1940, cxiv, 455-461. Google Scholar2. KOLMERRICHTERLYNCH JACEER: Lyophile complement fixation test for syphilis, Am. Jr. Clin. Path., 1938, viii, 522-528. CrossrefGoogle Scholar3. SHAKELL FF: An improved method of desiccation with some applications to biological problems, Am. Jr. Physiol., 1909, xxiv, 325-340. CrossrefGoogle Scholar4. ELSERTHOMASSTEFFEN WJBAGI: Desiccation of sera and other biological products (including microörganisms) in the frozen state with preservation of original qualities of products so treated, Jr. Immunol., 1935, xxviii, 433-473. Google Scholar5. GREAVESADAIR RIME: Simple method for the preservation of sera by desiccation in frozen state without use of refrigerants, Jr. Hyg., 1936, xxxvi, 507-513. Google Scholar6. FLOSDORFMUDD EWS: Procedure and apparatus for preservation in "lyophile" form of serum and other biological substances, Jr. Immunol., 1935, xxix, 389-425. Google Scholar7. MCGUINESSSTOKESMUDD ACJS: Clinical uses of the human serums preserved by lyophil process, Jr. Clin. Invest., 1937, xvi, 185-196. CrossrefGoogle Scholar8. STOKESMUDDARDDYEAGLEFLOSDORFLUCCHESI JSRLHEWP: Use of lyophile human serums for prevention and treatment in infectious diseases, Am. Jr. Dis. Child., 1934, xlviii, 1428-1430. Google Scholar9. MUDDFLOSDORFEAGLESTOKESMCGUINESS SEWHJAC: Preservation and concentration of human serums for clinical use, Jr. Am. Med. Assoc., 1936, cvii, 956-959. CrossrefGoogle Scholar10. BONDWRIGHT DDDG: Treatment of hemorrhage and traumatic shock by the intravenous use of lyophile serum, Ann. Surg., 1938, cvii, 500-510. CrossrefGoogle Scholar11. HUGHESMUDDSTRECKER JSEA: Reduction of increased intracranial pressure by concentrated solutions of human lyophile serum, Arch. Neurol. and Psychiat., 1938, xxxix, 1277-1287. CrossrefGoogle Scholar12. WRIGHTBONDHUGHES DGDDJ: Reduction of cerebrospinal fluid pressure by concentrated lyophile serum; further observations, Arch. Neurol. and Psychiat., 1938, xxxix, 1288-1293. CrossrefGoogle Scholar13. ALDRICHSTOKESKILLINGSWORTHMCGUINESS CAJWPAC: Concentrated human blood serum as diuretic in treatment of nephrosis; preliminary report Jr. Am. Med. Assoc., 1938, cxi, 129-133. CrossrefGoogle Scholar14. RAVDIN IS: Some recent advances in surgical therapeusis, Ann. Surg., 1939, cix, 321-333. CrossrefGoogle Scholar15. FLOSDORFMUDD EWS: Improved procedure and apparatus for preservation of sera, microörganisms and other substances—cryochem-process, Jr. Immunol., 1938, xxxiv, 469-490. Google Scholar16. GREAVESADAIR RIME: High-vacuum condensation drying of proteins from frozen state, Jr. Hyg., 1939, xxxix, 413-445. Google Scholar17. BAUERPICKELS JHEG: Apparatus for freezing and drying virus in large quantities under uniform conditions, Jr. Exper. Med., 1940, lxxi, 83-88. CrossrefGoogle Scholar18. LEDNUM JM: Theory of adsorption. Read before Industrial Gas Section, sixteenth annual convention and exhibition of Am. Gas Assoc., Atlantic City, N. J., (Oct. 30), 1934. Google Scholar19. LEVINSON SO: Personal communication. Google Scholar This content is PDF only. To continue reading please click on the PDF icon. Author, Article, and Disclosure InformationAffiliations: Dallas, Texas*Read at the annual meeting of the Texas Club of Internal Medicine, Dallas, February 16, 1940.From the Department of Pathology, Hospital Laboratory Division, Baylor University College of Medicine. Nextarticle Advertisement FiguresReferencesRelatedDetails Metrics Cited byThe plasma proteins and their fractionationPreservation of Bacteria by LyophilizationThe plasma proteins and their fractionationThe plasma proteins and their fractionationThe Plasma Proteins and Their FractionationThe Use of Desiccated Plasma in UrologyÜber die entgiftende Therapie im KindesalterSHORT ABSTRACTS OF SOME OTHER ADVANCES IN PEDIATRICSON THE NATURE OF THE SACCHAROID FRACTION OF HUMAN BLOOD 1 August 1940Volume 14, Issue 2Page: 201-214KeywordsAntibodiesBlood plasmaHospital medicinePathology and laboratory medicineResearch laboratoriesSafetyTemperature ePublished: 1 December 2008 Issue Published: 1 August 1940 Copyright & PermissionsCopyright, 1940, by The American College of PhysiciansPDF downloadLoading ...

A Study of the Sensitivity and Specificity of a Simplified Complement Fixation Test for Syphilis

A Study of the Sensitivity and Specificity of a Simplified Complement Fixation Test for Syphilis

A Simplified Method for the Preparation of an Antigen for use in the Complement Fixation Test for Syphilis

Journal Article A Simplified Method for the Preparation of an Antigen for use in the Complement Fixation Test for Syphilis Get access Fred Boerner, Fred Boerner From the Laboratories of the Graduate Hospital, University of Pennsylvania, Philadelphia, Pennsylvania Search for other works by this author on: Oxford Academic Google Scholar Charles A. Jones, Charles A. Jones From the Laboratories of the Graduate Hospital, University of Pennsylvania, Philadelphia, Pennsylvania Search for other works by this author on: Oxford Academic Google Scholar Marguerite Lukens Marguerite Lukens From the Laboratories of the Graduate Hospital, University of Pennsylvania, Philadelphia, Pennsylvania Search for other works by this author on: Oxford Academic Google Scholar American Journal of Clinical Pathology, Volume 9, Issue 3, May 1939, Pages 321–328, https://doi.org/10.1093/ajcp/9.3.321 Published: 03 November 2015

Serologic Tests in the Diagnosis of Syphilis

THE Laboratory Section of the American Public Health Association has been interested for a number of years in the development of standard methods for the complement-fixation test for syphilis. Dr. Gilbert,' as referee, directed attention to establishing printiples of technic, upon which leading -serologists agreed, and which, if followed, might be expected to yield dependable results. So many phases of the technic rested upon a purely empirical basis that more complete information regarding the fundamental relationships that govern the reactions between the different reagents of the test was essential to further progress in standardization. After extended study, it has been possible recently to describe methods which permit a quantitative determination of the reaction and thus provide a rational basis for standardization of the test.2-4 Accurate measurement of the changes in complement activity that occur under the different conditions of the test depends upon the determination of the amount of complement required in each instance to give 50 per cent hemolysis, as well as upon the fact that constant and directlv proportional relationships

The complement fixation test for syphilis applied to oxalated blood

The complement fixation test for syphilis applied to oxalated blood

Is the Sensitizing Effect of Cholesterol for the Antigen Used in the Complement-Fixation Test for Syphilis Due to the Contaminating Sterol, Ergosterol?

Abstract Cholesterol which is widely used to increase the sensitivity of antigens employed in the complement-fixation test for syphilis was for many years considered to be a chemically pure substance. Through the studies made recently, however, of cholesterol as a source of vitamin D, the antirachitic vitamin, it is now known that cholesterol, as ordinarily prepared, contains another sterol of an unsaturated and labile type of which ergosterol is the only known representative. Furthermore, it is because of this small amount of contaminating sterol that cholesterol, following irradiation with ultra-violet light, acquires antirachitic properties (1, 2, 3, 4). When ergosterol is removed, cholesterol is inert after irradiation (1, 2, 5, 6). Ergosterol, on the contrary, is very highly active. In view of these observations, Professor Knudson of the Albany Medical College suggested that it would be of interest to determine whether cholesterol, after removal of the contaminating sterol, would lose its property of sensitizing the antigen used in the complement-fixation test for syphilis just as it loses its property of being rendered antirachitic by irradiation with ultraviolet light.

The Technique and Results of a Complement Fixation Test for the Diagnosis of Infections with Endamoeba Histolytica

In previous communications (1, 2), the writer briefly described a complement fixation test which has been found useful in the diagnosis of infections with Endamoeba histolytica, the cause of amoebic dysentery, and has detailed the results obtained with this test. In this paper the technique of the test will be described, together with the results that have been obtained by its application in actual clinical practice, as well as experimentally, over a period of nearly two years, during which time many hundreds of individuals have been tested, the results being checked by microscopical and cultural examinations of the feces of the tested individuals, and by the clinical histories and symptoms of such individuals.Technique of the Test. The technique of this complement fixation test is practically the same as that used as the standard method for performing the complement fixation test for syphilis in the United States Army laboratories, fully described in the writer's book, “The Wassermann Test” (3), with suitable modifications made necessary by the difference in the character of the antigen used in this test.

Is the Sensitizing Effect of Cholesterol for the Antigen Used in the Complement-fixation Test for Syphilis Due to the Contaminating Sterol, Ergosterol?

Cholesterol as ordinarily prepared is now known to be contaminated with another sterol of an unsaturated and labile type of which ergosterol is the only known representative. Moreover, it is because of this contaminating sterol, ergosterol, that after irradiation with ultra-violet light, cholesterol acquires antirachitic properties. After removal of ergosterol, cholesterol is inert, following irradiation.Professor Knudson of the Albany Medical College very kindly offered a supply of relatively pure cholesterol and ergosterol and suggested that a comparative study be made to determine whether the sensitizing action which cholesterol has upon the antigen used in the complement-fixation test for syphilis is also due to the presence of the contaminating sterol, ergosterol.The antigen used was an alcoholic extract of dried, ether-extracted beef heart tissue. A comparison of the activity of the untreated antigen and of the antigen sensitized with the different samples of cholesterol and ergosterol used was made...

THE COMPLEMENT-FIXATION TEST FOR SYPHILIS

The following is a report of a series of investigations made to determine whether the patient's blood could be collected on filter paper, dried, and used in the complement-fixation test for syphilis. This procedure was attempted for the following reasons: less blood is necessary, which is especially important with children; the outfit for the collection of blood is simple and inexpensive; the risk of the specimen being spoiled by bacterial contamination or by the things which usually produce hemolysis might be eliminated, and in addition the specimens might be preserved for a long period of time with little deterioration. Noguchi, 1 in his modification of the Wassermann test, made use of the hemolytic amboceptor on such paper, with practical success. Can the substance present in the blood of persons with active syphilis dried on paper be preserved in a like manner? If so, can such a test be standardized so that

THE KOLMER COMPLEMENT FIXATION AS A SPECIFIC TEST FOR SYPHILIS

The complement fixation test for syphilis, initiated by Wassermann, has been made the subject of such exhaustive investigation and so many thorough statistical studies that it is practically impossible to find any new avenue of approach. Indeed, we find it difficult to say anything of this laboratory procedure without repeating, verbatim, the words of other workers. Lacking certain knowledge of the specific complement binding substances present in the tissues of the syphilitic individual, we cannot yet attain our goal in the diagnosis of syphilis. Facing the fact that antigens prepared from the causative organism are entirely unsatisfactory and that the tissue extract antigens in common use are in no possible way biologically specific; and, further, since the complement binding substance is most abundant at the height of the infection, as opposed to the antibody production in bacterial dis eases, it seems logical to consider the procedure in the light of

Experiments on the Removal of Hemagglutinin from Rabbit Antihuman Serum

Abstract In conducting the complement fixation test for syphilis with an antihuman hemolytic system, agglutination of the erythrocytes not infrequently interferes with hemolysis and the frequency and intensity of agglutination constitutes a drawback in the usefulness of the antihuman system. One of the sources of this agglutinin is the immune rabbit hemolytic serum and at the suggestion of Professor Kolmer, we have conducted a series of experiments with rabbit immune sera with the object of determining whether or not agglutinin for human cells may be removed without disturbing the hemolysin content. Professor Kolmer has been of the opinion that drying antihuman rabbit serum on paper after the method of Noguchi, resulted not only in facilitating the preservation and manipulation of the hemolysin but also in the deterioration of agglutinin without commensurate destruction of hemolysin and that this constituted an important reason for the use of paper amboceptor in complement fixation tests when an antihuman hemolytic system is employed.

The Complement Fixation Test for Syphilis: (Modified Wassermann.) Description of a Method at Present in Use at the Hygienic Laboratory

The Complement Fixation Test for Syphilis: (Modified Wassermann.) Description of a Method at Present in Use at the Hygienic Laboratory

Comparative Study of Different Antigens and of Different Temperatures of Incubation in the Wassermann Test

It is well recognized that widely different results may be obtained in complement-fixation tests for syphilis by the use of different antigens and by the employment of different incubation conditions, even with the same antigens. In a previous communication1 we have reported on Wassermann tests that were performed by 3 methods on 496 identical specimens from 477 patients. In the 1st method a cholesterin-reinforced antigen was employed and the 1st incubation was carried out at 37 C. In the 2nd method a simple alcoholic extract was used as antigen, with incubation also at 37 C. In the 3rd method this latter antigen was again employed, but the 1st incubation was carried out in the refrigerator for 4-24 hours. The last method proved more sensitive in the group of known syphilitics than the other procedures tested. Furthermore, a positive result thus obtained proved to be more trustworthy evidence of syphilis than did positive results obtained with the cholesterinized antigen and 1st incubation at 37 C. In the present paper we shall present the results of Wassermann tests performed by 6 methods on 501 identical specimens from 457 patients. In the 1st of these 6 methods a cholesterin-reinforced antigen was employed, and the 1st incubation was carried out at 37 C. In the 2nd method a simple alcoholic extract was used as antigen, with incubation also at 37 C. In the 3rd method an antigen prepared from the acetone-insoluble fraction of beef heart, after the method of Noguchi,2 was used, with incubation also at 37 C. Exact duplicates of these tests were prepared and incubated at 8 C, for 4 hours. Tests 4, 5, and 6 differ from Tests 1, 2, and 3, respectively, only in the temperature and length of time for the 1st incubation.

THE CONTROL OF VENEREAL DISEASES BY HEALTH DEPARTMENTS

H 'EALTH departments are created and financed to safeguard the public health. Each state and municipal health department has its own peculiar problem in deciding how it can most efficiently spend its appropriation. The character of the work of health departments has entirely changed in the past twenty-five years by reason of laboratory discoveries, and their application to public health measures. Probably the next generation will see even more striking changes. Notable achievements have been accomplished in health measures directed against diphtheria, tuberculosis, typhoid, yellow fever, malaria, hookworm, and other serious diseases. But two of the most common and dangerous of communicable diseases -syphilis and gonorrhoea-have been practically left untouched by health departments. There is, however, one striking exception, namely, the Health Department of the City of New York. Its sane and scientific work in venereal diseases merits the commendation of all medical men. This department was the pioneer in establishing free complement fixation tests for syphilis and gonorrhcea. The results of their laboratory work, and their methods of reporting cases, have demonstrated that there is a practical method which any state or city can apply to the control of v-enereal diseases. Personally, after many years' experience in laboratories, in hospitals, in the autopsy room, and in private practice, I have no hesitation in stating that the ravages of syphilis and gonorrhoea are far greater than those caused by diphtheria, scarlet fever and typhoid combined. Yet not so many years ago, these diseases occupied much of the attention of health authorities. It seems reasonable and proper to believe that the time has come when every state and municipal health department should, yearlv, spend some part of its appropriation in attempting the control of these diseases. The Worcester District Medical Society's Committee on Venereal Diseases, of which the writer is a member, recently sent out to its members a pamphlet, a part of which is quoted below:

A MODIFIED WASSERMANN

There are several undesirable features in the original Wassermann technic of the complement fixation test for syphilis, as well as in the Noguchi modification, some of which may be sources of error. In the original Wassermann technic the presence of variable amount of natural antisheep amboceptor, which is found in all human sera, may lead to error, unless removed by the tedious method of bringing in contact with sheep cells. Another objection to the original Wassermann is that unless a sheep is kept in the animal rooms of the laboratory it is sometimes difficult to secure corpuscles for the test, while the keeping of a sheep is more or less expensive, and in some laboratories well nigh impossible. In the Noguchi technic the use of a non-inactivated serum might, theoretically, be a source of error, although practically, owing to the small amount of serum used, probably is not. But

THE OCCURRENCE OF NATIVE ANTISHEEP AMBOCEPTOR IN HUMAN SERUM AND ITS IMPORTANCE IN THE PERFORMANCE OF THE WASSERMANN REACTION

That there occurs in certain human serums a substance which hemolyzes sheep corpuscles in the presence of complement has been known for a considerable time. It has been our purpose to find out how much the presence of this substance in serums which reacted positively to the Wassermann test could affect the final result of the reaction. In 1908 Bauer¹pointed out that use might be made of this substance — to which we shall refer as native antisheep amboceptor — in the complement fixation test for syphilis, and he suggested substituting this native amboceptor for the rabbit sheep amboceptor used in the Wassermann reaction. The Wassermann technic would be simplified in this way, as one of the reagents would be eliminated without endangering its accuracy. This procedure is known as the Bauer modification of the Wassermann reaction. Shortly after this, Bering²reported a series

SPECIFICITY OF THE NOGUCHI MODIFICATION OF THE WASSERMANN REACTION

Early in 1909 Noguchi¹brought to the attention of the medical profession a modification of the Wassermann system of complement fixation test for syphilis. It was in reality a new system for the detection of the so-called syphilitic antibodies, based on the Bordet-Gengou²phenomenon of complement fixation, as in the Wassermann method, but using an antihuman hemolytic system as indicator instead of the antisheep system of the original method. Although differing from the latter in technical application, this variation in the hemolytic indicator represents the fundamental difference between the two systems. Noguchi claimed for his method simplicity, greater facility of application, greater sensitiveness in the detection of the specific reacting bodies, and the possibility of so preparing the various reagents used in the test as to make it available for every laboratory worker or physician familiar with clinical laboratory methods. A glance at Noguchi's