complement fixation tests


Summary: Serologic tests based on inactivation of complement by the antigen-antibody complex (stage 1). Binding of free complement can be visualized by addition of a second antigen-antibody system such as red cells and appropriate red cell antibody (hemolysin) requiring complement for its completion (stage 2). Failure of the red cells to lyse indicates that a specific antigen-antibody reaction has taken place in stage 1. If red cells lyse, free complement is present indicating no antigen-antibody reaction occurred in stage 1.

Top Publications

  1. Naseem S, Meens J, Jores J, Heller M, Dubel S, Hust M, et al. Phage display-based identification and potential diagnostic application of novel antigens from Mycoplasma mycoides subsp. mycoides small colony type. Vet Microbiol. 2010;142:285-92 pubmed publisher
    ..This prototype test resulted in 100% diagnostic sensitivity and specificity. In comparison, the complement fixation test and the competitive ELISA performed with a diagnostic sensitivity of 70% and 60%, respectively...
  2. Bruderer U, Regalla J, Abdo E, Huebschle O, Frey J. Serodiagnosis and monitoring of contagious bovine pleuropneumonia (CBPP) with an indirect ELISA based on the specific lipoprotein LppQ of Mycoplasma mycoides subsp. mycoides SC. Vet Microbiol. 2002;84:195-205 pubmed
    ..The study revealed that the ELISA was both specific and sensitive for CBPP positive bovine sera and was shown also to be robust to harsh climatic conditions...
  3. Sacchini F, Naessens J, Awino E, Heller M, Hlinak A, Haider W, et al. A minor role of CD4+ T lymphocytes in the control of a primary infection of cattle with Mycoplasma mycoides subsp. mycoides. Vet Res. 2011;42:77 pubmed publisher
    ..The slightly higher mortality in the depleted group suggests a minor role of CD4+ T cells in control of CBPP. ..
  4. Marobela Raborokgwe C, Nicholas R, Ayling R, Bashiruddin J. Comparison of complement fixation test, immunoblotting, indirect ELISA, and competitive ELISA for detecting antibodies to Mycoplasma mycoides subspecies mycoides small colony (SC) in naturally infected cattle from the 1995 outbreak in Botswana. Onderstepoort J Vet Res. 2003;70:21-7 pubmed
    ..It became clearly evident from this comparative study that no single serological test was capable of detecting all animals affected by CBPP under natural field conditions of infection...
  5. Moncayo A, Hice C, Watts D, Travassos de Rosa A, Guzman H, Russell K, et al. Allpahuayo virus: a newly recognized arenavirus (arenaviridae) from arboreal rice rats (oecomys bicolor and oecomys paricola) in northeastern peru. Virology. 2001;284:277-86 pubmed publisher
    ..A predicted stable single hairpin secondary structure was identified in the intergenic region between GPC and NP. Details of the genetic organization of Allpahuayo virus are discussed...
  6. Adone R, Ciuchini F, Olsen S. Field validation of the use of RB51 as antigen in a complement fixation test to identify calves vaccinated with Brucella abortus RB51. Clin Diagn Lab Immunol. 2001;8:385-7 pubmed
  7. Amanfu W, Sediadie S, Masupu K, Raborokgwe M, Benkirane A, Geiger R, et al. Comparison between c-ELISA and CFT in detecting antibodies to Mycoplasma mycoides mycoides biotype SC in cattle affected by CBPP in Botswana. Ann N Y Acad Sci. 2000;916:364-9 pubmed
    ..The CFT was found to be slightly more sensitive than the c-ELISA, and this could be related to the stage of the disease. A long-term study comparing the progression of the disease with the two tests is, therefore, essential...
  8. Williams S, Scalarone G. Lack of antibody induction following skin testing with histolyn-CYL: ELISA determinations with Histoplasma capsulatum yeast and mycelial phase antigens. Diagn Microbiol Infect Dis. 1987;8:127-30 pubmed
  9. Raso T, Carrasco A, Silva J, Marvulo M, Pinto A. Seroprevalence of antibodies to Chlamydophila psittaci in zoo workers in Brazil. Zoonoses Public Health. 2010;57:411-6 pubmed publisher
    ..This seroepidemiological survey confirmed the need to adopt preventive measures to control avian chlamydiosis and protect the health of zoo workers in the country. ..

More Information


  1. Mellbye O, Natvig J. Evidence for immune complexes containing antibody to the pepsin site of IgG in rheumatoid synovial fluids. Clin Exp Immunol. 1971;8:889-99 pubmed
  2. Schubert E, Sachse K, Jores J, Heller M. Serological testing of cattle experimentally infected with Mycoplasma mycoides subsp. mycoides Small Colony using four different tests reveals a variety of seroconversion patterns. BMC Vet Res. 2011;7:72 pubmed publisher
    ..The methods used included an in-house complement fixation test (CFT), a commercially available CFT, a competitive antibody ELISA (cELISA) and the immunoblotting test (IBT). In addition, lung tissue samples were examined by culture...
  3. Tellez A, Martin A, Anda P, De la Fuente L, Benitez P, Garcia C, et al. Study of C. burnetii human and animal seroprevalence in a rural population in Madrid community. Eur J Epidemiol. 1989;5:444-6 pubmed
    ..An urban sample shows an 8.8% positivity. Animal seroprevalence in livestock located in the village under study is 76.6% in caprine and 17.7% in bovine. A high risk epidemiological situation is detected. ..
  4. Yoshida T, Kamiyama T, Okada T, Nakanishi K, Yokoo H, Kamachi H, et al. Alveolar echinococcosis of the liver in children. J Hepatobiliary Pancreat Sci. 2010;17:152-7 pubmed publisher
    ..Thus, screening examinations are essential for children in contaminated areas and, if a liver tumor is found on a screening examination and diagnosed as AEL, complete radical resection should be performed. ..
  5. Okada M, Asai T, Futo S, Mori Y, Mukai T, Yazawa S, et al. Serological diagnosis of enzootic pneumonia of swine by a double-sandwich enzyme-linked immunosorbent assay using a monoclonal antibody and recombinant antigen (P46) of Mycoplasma hyopneumoniae. Vet Microbiol. 2005;105:251-9 pubmed
    ..Therefore, this ELISA is a useful alternative to the CF test currently used for the diagnosis of EP. ..
  6. Henchal E, Henchal L, Schlesinger J. Synergistic interactions of anti-NS1 monoclonal antibodies protect passively immunized mice from lethal challenge with dengue 2 virus. J Gen Virol. 1988;69 ( Pt 8):2101-7 pubmed publisher
    ..These results may affect the design of recombinant dengue vaccines which may require the inclusion of serotype-specific antigenic domains...
  7. Aguirre N, Vanzini V, Torioni de Echaide S, Valentini B, De Lucca G, Aufranc C, et al. Antibody dynamics in Holstein Friesian heifers vaccinated with Brucella abortus strain 19, using seven serological tests. J Immunoassay Immunochem. 2002;23:471-8 pubmed
    ..5% positive animals at week 4 and then declined. Based on these data, the c-ELISA and FPA discriminated residual antibody activity due to vaccination more efficiently than the other tests. ..
  8. Burkhardt U, Zahn R, Hoffler U, Siegler K, Frilling B, Weber M, et al. Antibody levels against Chlamydia pneumoniae and outcome of roxithromycin therapy in patients with acute myocardial infarction. Results from a sub-study of the randomised Antibiotic Therapy in Acute Myocardial Infarction (ANTIBIO) trial. Z Kardiol. 2004;93:671-8 pubmed
    ..pneumoniae and treatment with roxithromycin did not influence these events, independently of the serostatus against C. pneumoniae. However, the power of this subgroup analysis was low to detect small but significant differences. ..
  9. Shepherd A, Hummitzsch D, Leman P, Swanepoel R, Searle L. Comparative tests for detection of plague antigen and antibody in experimentally infected wild rodents. J Clin Microbiol. 1986;24:1075-8 pubmed
    ..Plague fraction 1 antigen was detected in 16 of 34 bacteremic sera from M. coucha and M. natalensis. The threshold sensitivity of the ELISA was approximately 10(5) Y. pestis per ml. ..
  10. Persson K, Boman J. Comparison of five serologic tests for diagnosis of acute infections by Chlamydia pneumoniae. Clin Diagn Lab Immunol. 2000;7:739-44 pubmed
    ..The tests were equally sensitive in different age groups. Reinfections seemed to be rare. ..
  11. Yamashita T. [Diagnostic tests: Varicella-zoster virus]. Nihon Rinsho. 2005;63 Suppl 7:294-6 pubmed
  12. Parrott R, Kim H, Arrobio J, Hodes D, Murphy B, Brandt C, et al. Epidemiology of respiratory syncytial virus infection in Washington, D.C. II. Infection and disease with respect to age, immunologic status, race and sex. Am J Epidemiol. 1973;98:289-300 pubmed
  13. Chin C, Chen G, Sequeria F, Berry G, Siehr S, Bernstein D, et al. Clinical usefulness of a novel C1q assay to detect immunoglobulin G antibodies capable of fixing complement in sensitized pediatric heart transplant patients. J Heart Lung Transplant. 2011;30:158-63 pubmed publisher
    ..Avoiding only the donor antigens that would be recognized by the C1q assay may accelerate time to transplant by expansion of the donor pool and potentially allows transplantation of previously "incompatible" organs. ..
  14. Bradway D, Torioni de Echaide S, Knowles D, Hennager S, McElwain T. Sensitivity and specificity of the complement fixation test for detection of cattle persistently infected with Anaplasma marginale. J Vet Diagn Invest. 2001;13:79-81 pubmed
    ..Results of this study indicate that the CF test is ineffective for identifying cattle persistently infected with A. marginale and thus is inadequate for anaplasmosis regulatory and surveillance programs...
  15. Grønhaug T, Ghildyal P, Barsett H, Michaelsen T, Morris G, Diallo D, et al. Bioactive arabinogalactans from the leaves of Opilia celtidifolia Endl. ex Walp. (Opiliaceae). Glycobiology. 2010;20:1654-64 pubmed publisher
    ..Collectively, these results indicate that the arabinogalactan type II polymers present in the leaves of O. celtidifolia may be used to develop medical devices for regulating inflammatory processes. ..
  16. Calder J, Reddy G, Chieves L, Courtney C, Littell R, Livengood J, et al. Monitoring Babesia bovis infections in cattle by using PCR-based tests. J Clin Microbiol. 1996;34:2748-55 pubmed
    ..bovis ranged from 58 to 70% for a single determination, while the sensitivity of the CF test was only 6%. Both PCR-based and CF tests for B. bovis had high specificity values ranging from 96 to 100%...
  17. Stanford F. Virus antibody in synovial fluid of rheumatoid and control patients. Ann Rheum Dis. 1974;33:342-5 pubmed
  18. Hokputsa S, Harding S, Inngjerdingen K, Jumel K, Michaelsen T, Heinze T, et al. Bioactive polysaccharides from the stems of the Thai medicinal plant Acanthus ebracteatus: their chemical and physical features. Carbohydr Res. 2004;339:753-62 pubmed
    ..The presence of the rare monosaccharide 3-O-methylgalactose may indicate that this can be used as a chemotaxonomic marker. The traditional way of using this plant as a medical remedy appears to have a scientific basis. ..
  19. Halsey E, Rasnake M, Hospenthal D. Coccidioidomycosis of the male reproductive tract. Mycopathologia. 2005;159:199-204 pubmed
    ..One received fluconazole for prostatic disease, while one received surgery followed by itraconazole for testicular disease. After 12 months of therapy, both remain asymptomatic and have decreased antibody titers against C. immitis. ..
  20. Li M, Shi Z, Li C, Yu L. Combining complement fixation and luminol chemiluminescence for ultrasensitive detection of avian influenza A rH7N9. Analyst. 2016;141:2061-6 pubmed publisher
    ..It is anticipated that the luminol CL-CFT assay would be particularly suitable for investigation of small molecules, toxins, and short peptides. ..
  21. Kumba F, Claasen B, Petrus P. Apparent prevalence of dourine in the Khomas region of Namibia. Onderstepoort J Vet Res. 2002;69:295-8 pubmed
  22. Muskens J, van Zijderveld F, Eger A, Bakker D. Evaluation of the long-term immune response in cattle after vaccination against paratuberculosis in two Dutch dairy herds. Vet Microbiol. 2002;86:269-78 pubmed
    ..In conclusion, for a large number of vaccinated cattle, a long lasting interference is to be expected with the presently available immunodiagnostic methods for both bovine tuberculosis and paratuberculosis. ..
  23. Peter O, Dupuis G, Bee D, Luthy R, Nicolet J, Burgdorfer W. Enzyme-linked immunosorbent assay for diagnosis of chronic Q fever. J Clin Microbiol. 1988;26:1978-82 pubmed
    ..The reading is objective, and the test is simple to perform and more sensitive than the IFA and complement fixation tests. The ELISA is recommended for serologic evaluation of patients with chronic Q fever.
  24. Scharp D, al Khalaf S, al Muhanna M, Cheema R, Godana W. Use of mass vaccination with a reduced dose of REV 1 vaccine for Brucella melitensis control in a population of small ruminants. Trop Anim Health Prod. 1999;31:135-41 pubmed
    ..The conclusion from these findings is that mass vaccination with reduced dose Rev 1 administered by the subcutaneous route is a practical field strategy for control of Brucella melitensis. ..
  25. Filipowicz E. Diagnostic methods for detection of respiratory RNA viruses. Acta Microbiol Pol. 2002;51:13-21 pubmed
    ..Patients at greatest risk of acquiring nosocomial viral respiratory disease are children, patients with immunodeficiency and patients treated in intensive care. ..
  26. McDonald H. Diagnostic and therapeutic challenges. Retina. 2003;23:847-51 pubmed
  27. Sidibé C, Grosbois V, Thiaucourt F, Niang M, Lesnoff M, Roger F. Performance evaluation of two serological tests for contagious bovine pleuropneumonia (CBPP) detection in an enzootic area using a Bayesian framework. Trop Anim Health Prod. 2012;44:1233-8 pubmed publisher
    ..However, negative conditional sensitivity dependence between the two tests was detected, implying that to achieve maximum sensitivity, the two tests should be applied in parallel. ..
  28. Khan I, Wieler L, Melzer F, Gwida M, Santana V, de Souza M, et al. Comparative evaluation of three commercially available complement fixation test antigens for the diagnosis of glanders. Vet Rec. 2011;169:495 pubmed publisher
    ..1 per cent) for each antigen were calculated to be 95.88 and 99.48 (, 97.04 and 100 (CIDC), 100 and 76.33 per cent (USDA), respectively. Almost perfect agreement (0.96) was found between CFT using either or CIDC and IB. ..
  29. Baatrup G, Jonsson H, Sjoholm A, Sturfelt G, Svehag S. Complement fixation by solid phase immune complexes. Reduced capacity in SLE sera. J Clin Lab Immunol. 1988;26:73-9 pubmed
    ..The concentrations of serum properdin were never found to be low, which was in contrast to the finding of reduced properdin uptake by solid phase IC in most of the samples. ..
  30. Schepetkin I, Xie G, Jutila M, Quinn M. Complement-fixing activity of fulvic acid from Shilajit and other natural sources. Phytother Res. 2009;23:373-84 pubmed publisher
    ..These data provide a molecular basis to explain at least part of the beneficial therapeutic properties of Shilajit and other humic extracts. ..
  31. Nielsen K, Smith P, Yu W, Elmgren C, Halbert G, Nicoletti P, et al. Validation of a second generation competitive enzyme immunoassay (CELISA) for the diagnosis of brucellosis in various species of domestic animals. Vet Immunol Immunopathol. 2008;125:246-50 pubmed publisher
    ..Sufficient samples from cattle, swine, sheep and goats to validate the test were included based on OIE guidelines suggesting inclusion of a minimum of 300 positive and 1000 negative samples. ..
  32. Tittarelli M, Bonfini B, De Massis F, Giovannini A, Di Ventura M, Nannini D, et al. Brucella abortus strain RB51 vaccine: immune response after calfhood vaccination and field investigation in Italian cattle population. Clin Dev Immunol. 2008;2008:584624 pubmed publisher
  33. Hermentin K, Hassl A, Picher O, Aspöck H. Comparison of different serotests for specific Toxoplasma IgM-antibodies (ISAGA, SPIHA, IFAT) and detection of circulating antigen in two cases of laboratory acquired Toxoplasma infection. Zentralbl Bakteriol Mikrobiol Hyg A. 1989;270:534-41 pubmed
  34. Niemczuk K, Bednarek D. Changes in the peripheral leukocyte phenotype of calves in clinical cases of bronchopneumonia complicated with chlamydial co-infectious agent. Pol J Vet Sci. 2003;6:125-9 pubmed
    ..Effective recruitment from a reserve marginal pool of these cells into blood vessels and activation of bone marrow proliferation are probably the reason for their high circulating number. ..
  35. Garin Bastuji B, Hummel N, Gerbier G, Cau C, Pouillot R, Da Costa M, et al. Non specific serological reactions in the diagnosis of bovine brucellosis: experimental oral infection of cattle with repeated doses of Yersinia enterocolitica O:9. Vet Microbiol. 1999;66:223-33 pubmed
    ..entercolitica O:9 shedding observed in herds showing false positive serological reactions in brucellosis. ..
  36. Fang X, Jiang B, Wang X. Purification and partial characterization of an acidic polysaccharide with complement fixing ability from the stems of Avicennia marina. J Biochem Mol Biol. 2006;39:546-55 pubmed
    ..Traditionally, this mangrove plant is used for medicinal purposes and it appears to have some scientific applications...
  37. de Freitas Raso T, Seixas G, Guedes N, Pinto A. Chlamydophila psittaci in free-living Blue-fronted Amazon parrots (Amazona aestiva) and Hyacinth macaws (Anodorhynchus hyacinthinus) in the Pantanal of Mato Grosso do Sul, Brazil. Vet Microbiol. 2006;117:235-41 pubmed
    ..No birds had clinical signs suggestive of chlamydiosis. To the best of our knowledge, this is the first report on C. psittaci in free-living Blue-fronted Amazon parrots and Hyacinth macaws in Brazil...
  38. Rawlins M, Gerstner C, Hill H, Litwin C. Evaluation of a western blot method for the detection of Yersinia antibodies: evidence of serological cross-reactivity between Yersinia outer membrane proteins and Borrelia burgdorferi. Clin Diagn Lab Immunol. 2005;12:1269-74 pubmed
    ..burgdorferi, which can cause an oligoarthritis similar to reactive arthritis, the diagnosis of reactive arthritis should be based on clinical findings and complete serologic analysis of the potential causative infectious pathogens. ..
  39. Pappagianis D. Seeking a vaccine against Coccidioides immitis and serologic studies: expectations and realities. Fungal Genet Biol. 2001;32:1-9 pubmed
    ..Concurrent conduct of the two approaches should maximize the chances for success. ..
  40. Ludwig R, Recke A, Bieber K, Müller S, Marques A, Banczyk D, et al. Generation of antibodies of distinct subclasses and specificity is linked to H2s in an active mouse model of epidermolysis bullosa acquisita. J Invest Dermatol. 2011;131:167-76 pubmed publisher
    ..Furthermore, our data are the basis for future studies with the goal of identifying non-MHC EBA susceptibility genes. ..
  41. Sutherland S, Chen G, Sequeira F, Lou C, Alexander S, Tyan D. Complement-fixing donor-specific antibodies identified by a novel C1q assay are associated with allograft loss. Pediatr Transplant. 2012;16:12-7 pubmed publisher
    ..These data suggest that DSA with the ability to activate complement, as determined by this novel C1q assay, are associated with greater risk of acute rejection and allograft loss. ..
  42. López L, Valencia Y, Tobón A, Velasquez O, Santa C, Caceres D, et al. Childhood histoplasmosis in Colombia: Clinical and laboratory observations of 45 patients. Med Mycol. 2016;54:677-83 pubmed publisher
    ..was reactive in 95%, the M band of the precipitate was more commonly observed (81%), and the complement fixation tests were reactive in 88% of the patients...
  43. Díaz Bello Z, Zavala Jaspe R, Díaz Villalobos M, Mauriello L, Maekelt A, de Noya B. [A confirmatory diagnosis of antibodies anti-Trypanosoma cruzi in donors referred by blood banks in Venezuela]. Invest Clin. 2008;49:141-50 pubmed
    ..cruzi in the integral evaluation of blood donors, since many of them with antibodies anti-T. cruzi, have donated blood several times previous to diagnosis. ..
  44. Capparelli R, Alfano F, Amoroso M, Borriello G, Fenizia D, Bianco A, et al. Protective effect of the Nramp1 BB genotype against Brucella abortus in the water buffalo (Bubalus bubalis). Infect Immun. 2007;75:988-96 pubmed
    ..The higher basal level of the antibacterial protein Nramp1 most probably provides the BB animals with the possibility of controlling bacteria immediately after their entry inside the cell...
  45. Hartley C, Wilks C, Studdert M, Gilkerson J. Comparison of antibody detection assays for the diagnosis of equine herpesvirus 1 and 4 infections in horses. Am J Vet Res. 2005;66:921-8 pubmed
    ..To compare methods of detecting equine herpesvirus type 1 (EHV1)- and EHV4-specific antibodies in horse sera...
  46. Gwozdz J, Thompson K, Murray A, Reichel M, Manktelow B, West D. Comparison of three serological tests and an interferon-gamma assay for the diagnosis of paratuberculosis in experimentally infected sheep. Aust Vet J. 2000;78:779-83 pubmed
    ..The superior performance of the IFN-gamma assay in detecting infected sheep in this small experimental population warrants its further evaluation in a larger population of sheep naturally exposed to M a paratuberculosis. ..
  47. Morahan P, Thomson T, Kohl S, Murray B. Immune responses to labial infection of BALB/c mice with herpes simplex virus type 1. Infect Immun. 1981;32:180-7 pubmed
  48. Vikerfors T, Lindegren G, Grandien M, van der Logt J. Diagnosis of influenza A virus infections by detection of specific immunoglobulins M, A, and G in serum. J Clin Microbiol. 1989;27:453-8 pubmed
    ..IgM detection by the hemadsorption immunosorbent technique was found to be a valuable supplement for the diagnosis of influenza A in an early phase of the disease. ..
  49. Lavaroni O, Aguirre N, Vanzini V, Lugaresi C, Torioni de Echaide S. [Assessment of polymerase chain reaction (PCR) to diagnose brucellosis in a Brucella infected herd]. Rev Argent Microbiol. 2004;36:101-6 pubmed
    ..Serological test showed 100% sensitivity related to PCR. The specificity for CF, C-ELISA and I-ELISA was 100%, 99% and 95% respectively. PCR could be useful to identify Brucella biotypes and to complement serologic tests...
  50. Mathias L, Girio R, Duarte J. Serosurvey for antibodies against Brucella abortus and Leptospira interrogans in pampas deer from Brazil. J Wildl Dis. 1999;35:112-4 pubmed
    ..Antibodies against B. abortus were detected by plate agglutination, rose Bengal, and complement fixation tests; antibodies against Leptospira interrogans were detected by the microscopic agglutination test...
  51. Cavanaugh D, Elisberg B, Llewellyn C, Marshall J, Rust J, Williams J, et al. Plague immunization. V. Indirect evidence for the efficacy of plague vaccine. J Infect Dis. 1974;129:Suppl:S37-40 pubmed
  52. Hegazy Y, Elmonir W, Abdel Hamid N, Elbauomy E. Seroprevalence and "Knowledge, Attitudes and Practices" (KAPs) survey of endemic ovine brucellosis in Egypt. Acta Vet Scand. 2016;58:1 pubmed publisher
  53. Vernon S, Webb P. Recent vesicular stomatitis virus infection detected by immunoglobulin M antibody capture enzyme-linked immunosorbent assay. J Clin Microbiol. 1985;22:582-6 pubmed
    ..The capture IgM ELISA is applicable for the rapid detection of IgM antibody to VSV in cattle and horses and is a useful assay of recent infection. ..