CF Test Research Articles

Effects of lymphoid cells and serum on target cells in xenogeneic cell-culture systems.

SummarySpleen lymphocytes of rabbits immunized with KB cells brought about destruction of these cells in tissue culture. No such effect was seen with normal lymphocytes. Immune sera from these rabbits reacted with KB cells in CF, CT, and SGI tests. The reaction was found to be specific as these sera did not react with cells of human amnion or of hamster tumors induced by adenovirus type 12. Anti-KB serum did react, however, with cells derived from fetal human skin.

A Sensitive Neutralization Test for the Adeno-Associated Satellite Viruses

Abstract A highly sensitive neutralization test for the four satellite serotypes was developed by selecting particularly effective combinations of adenovirus helper and cell system. Neutralizing antibody titers were measured by inhibition of satellite replication as assayed in terms of complement-fixation-producing units (CFPU) of virus. Neutralizing antibody measured by this method was more type-specific, and the antibody titers were much higher than those obtained by the direct complement-fixing antibody test. Homotypic neutralizing antibody titers of 103 were common, and in some instances titers as high as 106 were observed. The neutralizing activity against satellite virus was found to reside in the λ globulin fraction of post-infection human and post-immunization rabbit sera. The frequency of individuals with neutralizing antibodies but without CF antibodies suggests that the CF antibody titer falls off more rapidly than the neutralizing titer. Both neutralization and CF tests revealed significant intertypic crossing, especially between types 2 and 3.

Comparative evaluation of tests in the diagnosis of pulmonary hydatid cysts

A comparative study of 3 immunological tests was carried out in 25 pulmonary hydatid cyst patients. In the pre-operative period, the mean percentage of positive reactors tested by Casoni's test, complement fixation test and tanned red cell haemagglutination test were 80%, 60% and 52% respectively. However, when all the 3 tests were performed, one or the other test was found to be positive in 88% of the cases. In the post-operative period the Casoni test remained positive in all the positive cases for 5 years, whereas the CF and TRCH tests became negative within 1 year.

Paracoccidioidomycosis (South American Blastomycosis) *

A study designed to determine the various clinical, epidemiologic, and diagnostic aspects of paracoccidioidomycosis was undertaken. Clinical studies revealed that most of the patients had pulmonary involvement either as the only manifestation of the disease (46.1%) or in combination with other manifestations (47.3%). Mucous-membrane lesions were second in importance (51.0%). More than 90.0% of the patients were over 30 years of age, all but one were males, and more than 80.0% worked in occupations related to contact with soil. It was found that 69.4% of the cases originated in the subtropical forest. Methods such as biopsies, direct (KOH) mounts, and cultures proved to be reliable diagnostic tools as they allowed observation, or isolation, or both, of Paracoccidiodes brasiliensis in more than 80.0% of the cases. Serologic procedures (complement-fixation [CF] and agar-gel immunodiffusion tests) were also found to be valuable diagnostic methods. At the time of admission, 79.4% of the patients had a positive CF test, and 92.3% gave precipitin bands. The results of the serologic tests were of great help in case-finding.

Comparison of Macrocomplement and Microcomplement Fixation Techniques Used in Fungus Serology

A comparative evaluation was performed on the micro- and macrocomplement fixation (CF) tests that are used as standard procedures in the serodiagnosis of blastomycosis, coccidioidomycosis, and histoplasmosis. Tests with 937 sera from suspected and culturally proven cases of these diseases against yeast-form antigens of Blastomyces dermatitidis and Histoplasma capsulatum and against the soluble antigens, coccidioidin and histoplasmin, revealed that the microtiters were within +/- 1 dilution of the macrotiters in 83 to 93% of the sera. Tests on randomly coded quality control sera revealed the microform of the CF test to be highly reproducible. Our studies indicate that the results obtained by the two tests have similar diagnostic and prognostic interpretations. Because of the sensitivity, reproducibility, economy, and ease of performance, the microtest is highly recommended for use in fungus serology.

Infections with viruses in patients hospitalized with acute respiratory illness, Stockholm 1963-1967.

During 1963–1967 a study was undertaken of 1128 patients hospitalized with acute respiratory illness. 53% of the patients had pneumonia and most of them were 30 years of age or older.Complement fixation (CF) tests were carried out against respiratory viruses, including adenovirus, influenza A and B viruses, parainfluenza viruses 1, 2, and 3, and respiratory syncytial virus, as well as a Bedsonia agent, and Mycoplasma pneumoniae (MP). Antilysin tests against β-haemolytic streptococci, Staphylococcus aureus and pneumococci, and CF tests against Haemophilus influenzae were also performed.A significant titre rise to viruses was noted in 27% of the patients. This overall incidence was almost the same in different age groups, whether or not the patients had pneumonia.Antibody responses to more than one viral antigen, multiple responses within the parainfluenza group excluded, were found in 9% of the positive cases. In patients with CF titre rises to viral antigens 59% of those with pneumonia and 28% of those without pneumonia also developed significant titre rises against bacteria and/or Bedsonia and/or MP.Patients with CF titre rise to viral antigens were analysed with respect to the seasonal incidence of infection and the clinical picture. In spite of the fact that the same investigator had examined all the patients, only minor variations in signs and symptoms were noted at admission between patients with signs of different single virus infections.

Properties of a "Herellea" Antigen Which Reacts with Ornithosis Antibody

Antigen. The characteristics of the HL organism of the Bacterium anitratum group used to prepare the antigen for the CF test have been described in detail by Shimizu and Bankowski [6]. Preparation of the bacterial CF antigen has also been described elsewhere [5]. Briefly, the HL bacteria were cultured on 50 tryptose agar plates for 18 hr at 37 C. Cells were harvested from the agar surface,1 pooled, washed in 0.85^o saline solution, and resuspended in 150 ml of

A Complement Fixation Test for the Study of Avian Mycoplasmas

A CF test was developed which was of use in identifying newly isolated strains of avian mycoplasmas. An unequivocal assignment of serotype was possible for about half of the antigens prepared from 2nd or 3rd broth passage. With others the reactions were insufficient to allow for positive identification. From preliminary investigation it appeared possible that the CF test might be used as a diagnostic aid by applying it to the detection of mycoplasmal antigens in tissues. The test was useful for quantitating the production of antigen in cultures.

Scrotal abnormalities in rams in tropical Queensland with particular reference to ovine brucellosis and its control.

SummaryClinical conditions affecting the reproductive organs were found affecting 17.2% of 2,332 rams. The following conditions were found: epididytimis 69.4%, atrophy 15.4%, varicocoele 9.5%, hernia 2.9%, cryptorchidism 1.2%, and hydrocoele 1.5%. The incidence of Brucella ovis antibodies as determined by the CF test in 2,332 rams from 12 flocks in north western Queensland was found to be 18.3%.The age distribution of ovine brucellosis among 1,772 rams whose ages were known was as follows: 1 year 2.1%, 2 years 10.4%, 3 years 15.9%, 4–7 years 21.8%, and over 7 years 24.4%. The age incidence of all the various clinical conditions as they affected the 1,772 rams was: 1 year 3.6%, 2 years 8.4%, 3 years 13.1%, 4–7 years 28.4%, and over 7 years 27.9%.Control of brucellosis in two flocks is reported and a possible means of control of this disease in Queensland is discussed.

An Evaluation Of Intraderaml Tests For The Screening Of Paragonimiasis By Comparing Antigens Variously Processed.

An evaluation of intradermal test in screening paragonimiasis was attempted by comparing antigens, filtered through Sephadex G 50, fractionated by DEAE Sephadex A 25, and V.B.S.(crude extract of adult worms) antigen on 402 school children. The sensitivity and false negative rates were compared with one another on results of two examinations for eggs, one with sputum and the other with stool. The purified antigens showed more intense skin reaction than the crude extract. The crude extrct(V.B.S.) appeared to have lower false negative rate than the purified even though the difference was not statistically significant. Intradermal test with V.B.S. antigen on 12,000 subjects was analysed to find some characteristics of the reaction. Three size groups were classified by egg detection rate; negative reaction, 20~50 mm(2) size group of which egg detection rate was 3.5% among 118 persons, intermediate, 60~100 mm(2) with 26.8% among 1,078, and positive reaction, larger than 100 mm(2) with 45.7% among 2,098 subjects. The distribution of wheal size by age for total population surveyed revealed that younger ages had smaller wheal sizes with larger proportion of negative reaction, which decreased when the age increased. Skin sensitization in a population seemed to occur gradually to and reached a maximum by the age of 13 year. The egg detection rate increased proportionately to wheal size for the intermediate group, however, it did not vary much by size for positive group(larger than 100 mm(2)). When the wheal size distributions were compared in three groups, egg positives, egg negatives, and the group whose sputa were not examined, curves of the two groups, egg positive and negative, did not show clear separation. This fact seems to indicate that there is a low sensitivity and specificity in both tests(sputum and skin). The stability of the intradermal test measured by a second survey done one year after the primary survey revealed that the intermediate size group had more marked unstability than the other groups, and the treated group had tendency to decrease in wheal size. The purification of antigen for intradermal test did not add any better means of screening for paragonimiasis, probably due to the nature of skin sensitization evoked by many elements of the parasite but not by a few selected components. The V.B.S. antigen may be valuable for screening a large population, yet the diagnosis can not be dependent entirely upon the test. The final diagnosis should be made by repeated sputum examinations in spaced time with careful history taking aided by CF test and X-ray examination if necessary.

Fluorescent antibody tests in pre-clinical experimental Mycobacterium johnei infections in cattle

Antibodies detectable by the fluorescent antibody test developed in animals experimentally infected with M. johnei orally, between 3 and 12 months after dosing. The FA test was found to be positive 3 times more often than the CF test up to 22 to 24 months after dosing, when the animals were killed and their infections assessed. The fluorescent antibody levels were shown to vary from month to month and in some cases weekly, but tests on individual sera were reproducible. The incidence of fluorescent antibodies in cattle which had not been experimentally infected with M. johnei but came from a similar source, was 8·5 per cent. The possible causes of such reactions are discussed.

A plaque neutralization test for determining mumps antibodies.

SummaryThe mumps plaque neutralization assay procedure was found to be a sensitive indicator of low levels of mumps antibodies. Undiluted serum can be studied for the presence of mumps antibodies and there is no problem of nonspecific inhibition. The levels of antibody 1 year after natural mumps are more stable by the PN method than by HAI or CF tests. This assay procedure is valuable in studying sera where low levels of antibody may be present, as after remote natural disease, or vaccination with attenuated mumps strains.

Yaba tumor virus. I. Studies on pathogenesis and immunity.

SummaryTumor development and CF antibody responses of monkeys inoculated s.c. and i.v. with various dilutions of Yaba tumor virus were investigated. Tumors developed at different times and at different sites with CF antibodies appearing soon after the development of tumors and attaining final CF titers comparable in all inoculated animals. No antigenic relationship between Yaba virus and monkey pox viruses was found by in vivo challenge, or between Yaba, monkey pox, and vaccinia virus by cross CF and HI tests. The development of viral antigen(s) in tumor cells removed from infected monkeys at weekly intervals was monitored by the FA technique and CF test. Although the FA technique detected the viral antigen earlier, both procedures were equally effective for detection of antigen in tumors removed 4 or more weeks postinfection.

Method for increasing the diagnostic capacity of the complement fixation test in some resporatory virus infections.

SummaryThe effect of adding dextran T 80 (av mol. wt. 80,000) in certain concentrations to the mixture of antigen, complement (C) and serum in the diagnostic CF test for RS and parainfluenza (P) virus infections has been investigated. Preliminary experiments showed that the diluent employed (Veronal buffered saline, VBS) had to be used with hypertonic salt concentrations to prevent C consumption from increasing when dextran was added to the test system. Sera from infants and children with respiratory infections were titrated in parallel CF tests using either regular VBS without dextran (CF-v) or hypertonic VBS supplemented with dextran (CF-d). In the majority of sera examined the CF-d titers were significantly higher than the corresponding CF-v titers. Furthermore, using dextran, a rise in titer in the second of a pair of sera was obtained, which was not found in regular CF tests without dextran. Addition of dextran was also able to increase the CF titers in tests with LCM antigen and immune sera from mice.

Comparative Antigenic Study of Besnoitia Jellisoni, B. Panamenis and Five Toxoplasma Gondii Isolates

Summary Besnoitia jellisoni and B. panamensis were compared antigenically with five Toxoplasma gondii isolates. These were found to be related antigenically as shown by the CF, IHA and IFA serologic tests. Not all of the T. gondii isolates revealed the same degree of related antigenicity to Besnoitia. The T. gondii RH antigen showed no antigenic crossing with the Besnoitia antiserum in the CF and IFA tests, but cross-reactions were observed when T. gondii RH antiserum was reacted with Besnoitia antigen. The other Toxoplasma isolates crossed antigenically with both species of Besnoitia in the CF test, and all the Toxoplasma isolates were found to cross with both Besnoitia species in the IHA test. The IHA test with absorbed sera revealed that of the Toxoplasma isolates, the T. gondii RH was antigenically the least related to the Besnoitia. The comparative antigenic studies of these two genera suggested that an antigenically related spectrum of organisms exists with the Besnoitia and the T. gondii RH isolate at opposite ends of the spectrum. The other Toxoplasma isolates were found to have strong antigenic similarities between the two extremes with the T. gondii VI-6 chicken isolate being closer to the Besnoitia and the T. gondii Taylor isolate closer to the T. gondii RH isolate. These studies also showed that there are antigenic differences among the Toxoplasma group.

A New Serologic Test for Adenovirus Infection

Summary A new serologic test for adenovirus infection in man has been described. The SAFA test utilizes an adenovirus tissue culture antigen which is dried upon cellulose acetate discs, incubated with human serum at a single dilution of 1:25 and then reacted with fluorescein-conjugated antihuman globulin antiserum. Fluorescence is read objectively in a fluorometer. The SAFA test is comparable to the CF test for adenoviruses in that it is group-specific rather than type-specific. Twenty-nine of 32 patients with significant CF antibody responses were SAFA test positive. Eleven of 36 CF negative patients were SAFA positive. It would appear that the SAFA test is more sensitive than the CF test since 7 of the 11 had other evidence for adenoviral infection. Other advantages of the SAFA test are the use of a single dilution of serum, the requirement for very small amounts of serum and the stability of the antigens used.

A whey complement fixation test. Its relation to whey agglutination and isolation of Brucella abortus from the milk of individual cows.

SUMMARYTwo hundred and ninety-three milk ring test-positive individual milk samples were examined by whey complement fixation and agglutination tests and the results were compared with the isolation of B. abortus by cultures made from the milk.There was a closer correlation between whey CF tests and isolation of the organism than between whey AG tests and isolation. Brucella organisms were not isolated from 116 MRT-positive milk samples when the whey CF titre was < 1/10; whereas 9/104 (9%) isolations were made from milk when the whey AG titre was < 1/20.Because of its closer correlation with culture results it is suggested that the whey CF test would be valuable in an eradication programme for the detection of infected udders.The limitations of the milk ring test for the examination of individual milk samples are emphasized and it should be used only as a preliminary screening test. In the individual milk samples examined, 32% of the MRT-positives showed no evidence of infection by either the whey AG, whey CF, or culture tests.

Antigens of Rickettsia Prowazeki

Summary Antigenic activities of intact rickettsiae, cell walls, protoplasm and soluble antigens of Rickettsia prowazeki were compared on the basis of weight in complement fixation and mouse protection tests. All of the materials were active in both systems. The protoplasm antigens showed the greatest specificity and heat lability in the CF test. Cell walls and soluble antigens as well as intact rickettsiae contained more heat-stable CF antigens and tended to be more active than protoplasm in the mouse protection test although differences in this activity were not statistically significant. Heating the antigens provided evidence of dissociation between heat-labile mouse-protecting antigens and heat-stable CF antigens.

Characteristics of and relationship between C particles and intracisternal A particles in cloned cell strains.

Four murine tissue culture cell strains, which originated by cloning from one common cell of subcutaneous connective tissue origin, were examined for the presence of virus by electron microscopy and complement fixation techniques. The relative distribution of C particles and intracisternal A particles was determined. Thereafter, the characteristics of and relationship between A- and C-type particles were investigated. Cell extracts were passaged onto virus-free Swiss and C3Hf mouse embryo tissue cultures; CF and EM tests were again made to investigate the infective capacities of the various particle types. Although C particles were found in passage cultures exposed to extracts from the C particle-bearing strains, no A particles were found in any passage culture. These results indicate that the intracisternal A particle was neither infective nor developmentally associated with the C particle. A positive CF test was correlated with the presence of morphologically detectable C particles and was independent of the concentration of A particles.

螢光抗体間接法による日本脳炎ウイルス抗体の一測定法

The indirect method of immunofluorescence was used to titrate the antibody (Fluorescent antibody, FA) response in patients of Japanese encephalitis, after confirming that they had Japanese encephalitis by complement fixation test and hemagglutination inhibition test.The fluorescent antibody response patterns of 53 patients were studied. FA was found to appear early in the course of the disease, and tended to increase through the second or third post-onset week, although a few cases had maximal FA titer in the early stage of infection.The criterion for the serological diagnosis of Japanese encephalitis using indirect FA test was tentatively established as follow: more than 4-fold conversion should be observed between acute and convalescent specimens the latter titer being ≥1:80, or the serum at any stage should have a titer ≥1:160.According to the above criterion, 52 of 53 patients were positively diagnosed by the FA test. On the other hand, a diagnosis of Japanese encephalitis was made 51 cases out of 53 using HI test, and 36 cases out of 51 using CF test.Further analysis of the using various serological methods showed positive diagnosis were obtaind by HI test in 50% of patients, by FA test 30%, and by CF test 10% during the first 10 days of illness, by HI test was 90%, by FA test 75%, by CF test 30% during the first 20 days of illness.It is to be noted that FA test provide us a rapid reliable serological method for diagnosis. It takes about 200 minutes to examine one batch of 10 specimens with this test.Maximal FA titer obtained with case of inapparent Japanese encephalitis virus infection did not exceed 1:80, and those of normal cases 1:40 regardless their history of vaccination.