PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of jcmPermissionsJournals.ASM.orgJournalJCM ArticleJournal InfoAuthorsReviewers
 
J Clin Microbiol. 1994 October; 32(10): 2382–2386.
PMCID: PMC264070

PCR for capsular typing of Haemophilus influenzae.

Abstract

A PCR method for the unequivocal assignment of Haemophilus influenzae capsular type (types a to f) was developed. PCR primers were designed from capsule type-specific DNA sequences cloned from the capsular gene cluster of each of the six capsular types. PCR product was amplified only from the capsular type for which the primers were designed. Product was confirmed by using either an internal oligonucleotide or restriction endonuclease digestion. A total of 172 H. influenzae strains of known capsular type (determined genetically) comprising all capsular types and noncapsulate strains were tested by PCR capsular typing. In all cases the PCR capsular type corresponded to the capsular genotype determined by restriction fragment length polymorphism analysis of the cap region. When used in conjunction with PCR primers derived from the capsular gene bexA, capsulate, noncapsulate, and capsule-deficient type b mutant strains could be differentiated. PCR capsular typing overcomes the problems of cross-reaction and autoagglutination associated with the serotyping of H. influenzae strains. The rapid and unequivocal capsular typing method that is described will be particularly important for typing invasive H. influenzae strains isolated from recipients of H. influenzae type b vaccine.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Adams WG, Deaver KA, Cochi SL, Plikaytis BD, Zell ER, Broome CV, Wenger JD. Decline of childhood Haemophilus influenzae type b (Hib) disease in the Hib vaccine era. JAMA. 1993 Jan 13;269(2):221–226. [PubMed]
  • Anderson P, Johnston RB, Jr, Smith DH. Human serum activities against Hemophilus influenzae, type b. J Clin Invest. 1972 Jan;51(1):31–38. [PMC free article] [PubMed]
  • Catlin BW, Bendler JW, 3rd, Goodgal SH. The type b capsulation locus of Haemophilus influenzae: map location and size. J Gen Microbiol. 1972 May;70(3):411–422. [PubMed]
  • Falla TJ, Dobson SR, Crook DW, Kraak WA, Nichols WW, Anderson EC, Jordens JZ, Slack MP, Mayon-White D, Moxon ER. Population-based study of non-typable Haemophilus influenzae invasive disease in children and neonates. Lancet. 1993 Apr 3;341(8849):851–854. [PubMed]
  • Kroll JS, Ely S, Moxon ER. Capsular typing of Haemophilus influenzae with a DNA probe. Mol Cell Probes. 1991 Oct;5(5):375–379. [PubMed]
  • Kroll JS, Hopkins I, Moxon ER. Capsule loss in H. influenzae type b occurs by recombination-mediated disruption of a gene essential for polysaccharide export. Cell. 1988 May 6;53(3):347–356. [PubMed]
  • Kroll JS, Loynds B, Brophy LN, Moxon ER. The bex locus in encapsulated Haemophilus influenzae: a chromosomal region involved in capsule polysaccharide export. Mol Microbiol. 1990 Nov;4(11):1853–1862. [PubMed]
  • Kroll JS, Loynds BM, Moxon ER. The Haemophilus influenzae capsulation gene cluster: a compound transposon. Mol Microbiol. 1991 Jun;5(6):1549–1560. [PubMed]
  • Kroll JS, Zamze S, Loynds B, Moxon ER. Common organization of chromosomal loci for production of different capsular polysaccharides in Haemophilus influenzae. J Bacteriol. 1989 Jun;171(6):3343–3347. [PMC free article] [PubMed]
  • Musser JM, Kroll JS, Granoff DM, Moxon ER, Brodeur BR, Campos J, Dabernat H, Frederiksen W, Hamel J, Hammond G, et al. Global genetic structure and molecular epidemiology of encapsulated Haemophilus influenzae. Rev Infect Dis. 1990 Jan-Feb;12(1):75–111. [PubMed]
  • Salvi RJ, Ahroon W, Saunders SS, Arnold SA. Evoked potentials: computer-automated threshold-tracking procedure using an objective detection criterion. Ear Hear. 1987 Jun;8(3):151–156. [PubMed]
  • Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PubMed]
  • Shively RG, Shigei JT, Peterson EM, de la Maza LM. Typing of Haemophilus influenzae by coagglutination and conventional slide agglutination. J Clin Microbiol. 1981 Dec;14(6):706–708. [PMC free article] [PubMed]
  • Turk DC. The pathogenicity of Haemophilus influenzae. J Med Microbiol. 1984 Aug;18(1):1–16. [PubMed]
  • van Ketel RJ, de Wever B, van Alphen L. Detection of Haemophilus influenzae in cerebrospinal fluids by polymerase chain reaction DNA amplification. J Med Microbiol. 1990 Dec;33(4):271–276. [PubMed]
  • Wallace RJ, Jr, Musher DM, Septimus EJ, McGowan JE, Jr, Quinones FJ, Wiss K, Vance PH, Trier PA. Haemophilus influenzae infections in adults: characterization of strains by serotypes, biotypes, and beta-lactamase production. J Infect Dis. 1981 Aug;144(2):101–106. [PubMed]
  • Zwahlen A, Kroll JS, Rubin LG, Moxon ER. The molecular basis of pathogenicity in Haemophilus influenzae: comparative virulence of genetically-related capsular transformants and correlation with changes at the capsulation locus cap. Microb Pathog. 1989 Sep;7(3):225–235. [PubMed]
  • Zwahlen A, Winkelstein JA, Moxon ER. Surface determinants of Haemophilus influenzae pathogenicity: comparative virulence of capsular transformants in normal and complement-depleted rats. J Infect Dis. 1983 Sep;148(3):385–394. [PubMed]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)