%0 Journal Article %1 falla_pcr_1994 %A Falla, T J %A Crook, D W %A Brophy, L N %A Maskell, D %A Kroll, J S %A Moxon, E R %D 1994 %J Journal of Clinical Microbiology %K Bacterial Bacterial, Base Capsules, Chain Data, Genotype, Haemophilus Molecular Polymerase Reaction Sequence Sequence, influenzae, {DNA,} %N 10 %P 2382--6 %R 7814470 %T PCR for capsular typing of Haemophilus influenzae %U http://www.ncbi.nlm.nih.gov/pubmed/7814470 %V 32 %X 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.

@article{falla_pcr_1994, 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.}, added-at = {2011-03-11T10:05:34.000+0100}, author = {Falla, T J and Crook, D W and Brophy, L N and Maskell, D and Kroll, J S and Moxon, E R}, biburl = {https://www.bibsonomy.org/bibtex/29e1489531a68704ad13d943c35392723/jelias}, doi = {7814470}, interhash = {627b7ea175958ea6dc63800e0777b8dd}, intrahash = {9e1489531a68704ad13d943c35392723}, issn = {0095-1137}, journal = {Journal of Clinical Microbiology}, keywords = {Bacterial Bacterial, Base Capsules, Chain Data, Genotype, Haemophilus Molecular Polymerase Reaction Sequence Sequence, influenzae, {DNA,}}, month = oct, note = {{PMID:} 7814470}, number = 10, pages = {2382--6}, timestamp = {2011-03-11T10:06:09.000+0100}, title = {{PCR} for capsular typing of Haemophilus influenzae}, url = {http://www.ncbi.nlm.nih.gov/pubmed/7814470}, volume = 32, year = 1994 }