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. 1981 Apr;13(4):681–687. doi: 10.1128/jcm.13.4.681-687.1981

Minimum number of bacteria needed for antigen detection by counterimmunoelectrophoresis: in vivo and in vitro studies.

J C Fung, K Wicher
PMCID: PMC273859  PMID: 7014618

Abstract

Threshold concentrations of Streptococcus pneumoniae type 3, Haemophilus influenzae type b, and Streptococcus sp. group B type Ib required for positive counterimmunoelectrophoresis reactions were determined in vivo and in vitro. Animals were infected intraperitoneally with various concentrations of microorganisms: adult mice with S. pneumoniae, suckling rats with H. influenzae, and 3-week-old mice with Streptococcus sp. group B. At 24 h after infection a minimum blood concentration of 10(3) colony-forming units (CFU)/ml was needed for S. pneumoniae or H. influenzae before antigen was detected in the serum. A minimum concentration of 10(6) CFU/ml was needed for Streptococcus sp. group B at 10 h after infection. Larger threshold concentrations (10(4) CFU/ml for S. pneumoniae, 10(5) CFU/ml for H. influenzae, and 10(7) CFU/ml for Streptococcus) were required in broth-grown cultures before cell-free antigens could be demonstrated by counterimmunoelectrophoresis in the medium. Marked levels of antigen release by group B streptococci were observed as the cultures entered early stationary phase. This study provides evidence of a long-accepted, though poorly substantiated, hypothesis that a threshold concentration of microorganism is necessary before counterimmunoelectrophoresis reactions become positive. Counterimmunoelecrophoresis results for clinical specimens should be interpreted cautiously in light of this evidence.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson P., Pitt J., Smith D. H. Synthesis and release of polyribophosphate by Haemophilus influenzae type b in vitro. Infect Immun. 1976 Feb;13(2):581–589. doi: 10.1128/iai.13.2.581-589.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anhalt J. P., Yu P. K. Counterimmunoelectrophoresis of pneumococcal antigens:improved sensitivity for the detection of types VII and XIV. J Clin Microbiol. 1975 Dec;2(6):510–515. doi: 10.1128/jcm.2.6.510-515.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anthony B. F. Immunity to the group B streptococci: interaction of serum and macrophages with types Ia, Ib, and Ic. J Exp Med. 1976 May 1;143(5):1186–1198. doi: 10.1084/jem.143.5.1186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baltimore R. S., Kasper D. L., Baker C. J., Goroff D. K. Antigenic specificity of opsonophagocytic antibodies in rabbit anti-sera to group B streptococci. J Immunol. 1977 Feb;118(2):673–678. [PubMed] [Google Scholar]
  5. Barrett-Connor E. Bacterial infection and sickle cell anemia. An analysis of 250 infections in 166 patients and a review of the literature. Medicine (Baltimore) 1971 Mar;50(2):97–112. [PubMed] [Google Scholar]
  6. Colding H., Lind I. Counterimmunoelectrophoresis in the diagnosis of bacterial meningitis. J Clin Microbiol. 1977 Apr;5(4):405–409. doi: 10.1128/jcm.5.4.405-409.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Coonrod J. D., Drennan D. P. Pneumococcal pneumonia: capsular polysaccharide antigenemia and antibody responses. Ann Intern Med. 1976 Mar;84(3):254–260. doi: 10.7326/0003-4819-84-3-254. [DOI] [PubMed] [Google Scholar]
  8. Coonrod J. D., Rytel M. W. Detection of type-specific pneumococcal antigens by counterimmunoelectrophoresis. I. Methodology and immunologic properties of pneumococcal antigens. J Lab Clin Med. 1973 May;81(5):770–777. [PubMed] [Google Scholar]
  9. Damask L. J., Montoya O., Axelrod J. L. Rapid slide agglutination test for Lancefield grouping of streptococci. Arch Pathol Lab Med. 1979 Aug;103(9):456–458. [PubMed] [Google Scholar]
  10. Dossett J. H., Williams R. C., Jr, Quie P. G. Studies on interaction of bacteria, serum factors and polymorphonuclear leukocytes in mothers and newborns. Pediatrics. 1969 Jul;44(1):49–57. [PubMed] [Google Scholar]
  11. Drexhage H. A., van der Gaag R. D., Namavar F. Nitroblue tetrazolium-dye reduction by rat peritoneal macrophages during the uptake of Diplococcus pneumoniae, type VI. Antonie Van Leeuwenhoek. 1978;44(3-4):377–389. doi: 10.1007/BF00394314. [DOI] [PubMed] [Google Scholar]
  12. Edwards E. A., Muehl P. M., Peckinpaugh R. O. Diagnosis of bacterial meningitis by counterimmunoelectrophoresis. J Lab Clin Med. 1972 Sep;80(3):449–454. [PubMed] [Google Scholar]
  13. El-Refaie M., Dulake C. Counter-current immunoelectrophoresis for the diagnosis of pneumococcal chest infection. J Clin Pathol. 1975 Oct;28(10):801–806. doi: 10.1136/jcp.28.10.801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ellis E. F., Smith R. T. The role of the spleen in immunity. With special reference to the post-splenectomy problem in infants. Pediatrics. 1966 Jan;37(1):111–119. [PubMed] [Google Scholar]
  15. Feldman W. E. Effect of prior antibiotic therapy on concentrations of bacteria in CSF. Am J Dis Child. 1978 Jul;132(7):672–674. doi: 10.1001/archpedi.1978.02120320032006. [DOI] [PubMed] [Google Scholar]
  16. Feldman W. E. Relation of concentrations of bacteria and bacterial antigen in cerebrospinal fluid to prognosis in patients with bacterial meningitis. N Engl J Med. 1977 Feb 24;296(8):433–435. doi: 10.1056/NEJM197702242960806. [DOI] [PubMed] [Google Scholar]
  17. Fossieck B., Jr, Fedorko J. Counterimmunoelectrophoresis of blood cultures. Temporal Relationship of positive Gram stain to positive counterimmunoelectrophoresis. Am J Clin Pathol. 1979 Mar;71(3):326–329. doi: 10.1093/ajcp/71.3.326. [DOI] [PubMed] [Google Scholar]
  18. Granoff D. M., Congeni B., Baker R., Jr, Ogra P., Nankervis G. A. Countercurrent immunoelectrophoresis in the diagnosis of Haemophilus influenzae type b infection: relationship of detection of capsular antigen to age, antibody response, and therapy. Am J Dis Child. 1977 Dec;131(12):1357–1362. doi: 10.1001/archpedi.1977.02120250039006. [DOI] [PubMed] [Google Scholar]
  19. Hill H. R., Riter M. E., Menge S. K., Johnson D. R., Matsen J. M. Rapid identification of group B streptococci by counterimmunoelectrophoresis. J Clin Microbiol. 1975 Feb;1(2):188–191. doi: 10.1128/jcm.1.2.188-191.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ingram D. L., Anderson P., Smith D. H. Countercurrent immunoelectrophoresis in the diagnosis of systemic diseases caused by Hemophilus infleunzae type b. J Pediatr. 1972 Dec;81(6):1156–1159. doi: 10.1016/s0022-3476(72)80252-x. [DOI] [PubMed] [Google Scholar]
  21. Kenny G. E., Wentworth B. B., Beasley R. P., Foy H. M. Correlation of circulating capsular polysaccharide with bacteremia in pneumococcal pneumonia. Infect Immun. 1972 Oct;6(4):431–437. doi: 10.1128/iai.6.4.431-437.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lancefield R. C., McCarty M., Everly W. N. Multiple mouse-protective antibodies directed against group B streptococci. Special reference to antibodies effective against protein antigens. J Exp Med. 1975 Jul 1;142(1):165–179. doi: 10.1084/jem.142.1.165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mathews J. H., Klesius P. H., Zimmerman R. A. Opsonin system of the group B streptococcus. Infect Immun. 1974 Dec;10(6):1315–1320. doi: 10.1128/iai.10.6.1315-1320.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Merritt K., Treadwell T. L., Jacobs N. J. Rapid recognition of group B streptococci by pigment production and counterimmunoelectrophoresis. J Clin Microbiol. 1976 Mar;3(3):287–290. doi: 10.1128/jcm.3.3.287-290.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. O'Reilly R. J., Anderson P., Ingram D. L., Peter G., Smith D. H. Circulating polyribophosphate in Hemophilus influenzae, type b meningitis. Correlation with clinical course and antibody response. J Clin Invest. 1975 Oct;56(4):1012–1022. doi: 10.1172/JCI108148. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Shackelford P. G., Campbell J., Feigin R. D. Countercurrent immunoelectrophoresis in the evaluation of childhood infections. J Pediatr. 1974 Oct;85(4):478–481. doi: 10.1016/s0022-3476(74)80448-8. [DOI] [PubMed] [Google Scholar]
  27. Siegel J. D., McCracken G. H., Jr Detection of group B streptococcal antigens in body fluids of neonates. J Pediatr. 1978 Sep;93(3):491–492. doi: 10.1016/s0022-3476(78)81174-3. [DOI] [PubMed] [Google Scholar]
  28. Summary of the workshop on perinatal infections due to group B Streptococcus. J Infect Dis. 1977 Jul;136(1):137–152. doi: 10.1093/infdis/136.1.137. [DOI] [PubMed] [Google Scholar]
  29. Weller P. F., Smith A. L., Anderson P., Smith D. H. The role of encapsulation and host age in the clearance of Haemophilus influenzae bacteremia. J Infect Dis. 1977 Jan;135(1):34–41. doi: 10.1093/infdis/135.1.34. [DOI] [PubMed] [Google Scholar]

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