This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Falla, T J Articles by Moxon, E R Search for Related Content PubMed PubMed Citation Articles by Falla, T J Articles by Moxon, E R

PCR for capsular typing of Haemophilus influenzae.

Public Health Laboratory, University of Oxford, United Kingdom.

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.

This article has been cited by other articles:

• Garcia-Cobos, S., Campos, J., Cercenado, E., Roman, F., Lazaro, E., Perez-Vazquez, M., de Abajo, F., Oteo, J. (2008). Antibiotic Resistance in Haemophilus influenzae Decreased, except for {beta}-Lactamase-Negative Amoxicillin-Resistant Isolates, in Parallel with Community Antibiotic Consumption in Spain from 1997 to 2007. Antimicrob. Agents Chemother. 52: 2760-2766 [Abstract] [Full Text]  
• Ladhani, S, Slack, M P, Heys, M, White, J, Ramsay, M E (2008). Fall in Haemophilus influenzae serotype b (Hib) disease following implementation of a booster campaign. Arch. Dis. Child. 93: 665-669 [Abstract] [Full Text]  
• Benson, R., Tondella, M. L., Bhatnagar, J., Carvalho, M. d. G. S., Sampson, J. S., Talkington, D. F., Whitney, A. M., Mothershed, E., McGee, L., Carlone, G., McClee, V., Guarner, J., Zaki, S., Dejsiri, S., Cronin, K., Han, J., Fields, B. S. (2008). Development and Evaluation of a Novel Multiplex PCR Technology for Molecular Differential Detection of Bacterial Respiratory Disease Pathogens. J. Clin. Microbiol. 46: 2074-2077 [Abstract] [Full Text]  
• Sill, M. L., Tsang, R. S. W. (2008). Antibiotic Susceptibility of Invasive Haemophilus influenzae Strains in Canada. Antimicrob. Agents Chemother. 52: 1551-1552 [Abstract] [Full Text]  
• Erwin, A. L., Sandstedt, S. A., Bonthuis, P. J., Geelhood, J. L., Nelson, K. L., Unrath, W. C. T., Diggle, M. A., Theodore, M. J., Pleatman, C. R., Mothershed, E. A., Sacchi, C. T., Mayer, L. W., Gilsdorf, J. R., Smith, A. L. (2008). Analysis of Genetic Relatedness of Haemophilus influenzae Isolates by Multilocus Sequence Typing. J. Bacteriol. 190: 1473-1483 [Abstract] [Full Text]  
• Starner, T. D., Shrout, J. D., Parsek, M. R., Appelbaum, P. C., Kim, G. (2008). Subinhibitory Concentrations of Azithromycin Decrease Nontypeable Haemophilus influenzae Biofilm Formation and Diminish Established Biofilms. Antimicrob. Agents Chemother. 52: 137-145 [Abstract] [Full Text]  
• Mukundan, D., Ecevit, Z., Patel, M., Marrs, C. F., Gilsdorf, J. R. (2007). Pharyngeal Colonization Dynamics of Haemophilus influenzae and Haemophilus haemolyticus in Healthy Adult Carriers. J. Clin. Microbiol. 45: 3207-3217 [Abstract] [Full Text]  
• Satola, S. W., Collins, J. T., Napier, R., Farley, M. M. (2007). Capsule Gene Analysis of Invasive Haemophilus influenzae: Accuracy of Serotyping and Prevalence of IS1016 among Nontypeable Isolates. J. Clin. Microbiol. 45: 3230-3238 [Abstract] [Full Text]  
• Garcia-Cobos, S., Campos, J., Lazaro, E., Roman, F., Cercenado, E., Garcia-Rey, C., Perez-Vazquez, M., Oteo, J., de Abajo, F. (2007). Ampicillin-Resistant Non-{beta}-Lactamase-Producing Haemophilus influenzae in Spain: Recent Emergence of Clonal Isolates with Increased Resistance to Cefotaxime and Cefixime. Antimicrob. Agents Chemother. 51: 2564-2573 [Abstract] [Full Text]  
• Maaroufi, Y., De Bruyne, J.-M., Heymans, C., Crokaert, F. (2007). Real-Time PCR for Determining Capsular Serotypes of Haemophilus influenzae. J. Clin. Microbiol. 45: 2305-2308 [Abstract] [Full Text]  
• Zhou, J., Law, D. K. S., Sill, M. L., Tsang, R. S. W. (2007). Nucleotide Sequence Diversity of the bexA Gene in Serotypeable Haemophilus influenzae Strains Recovered from Invasive Disease Patients in Canada. J. Clin. Microbiol. 45: 1996-1999 [Abstract] [Full Text]  
• Cowgill, K. D., Ndiritu, M., Nyiro, J., Slack, M. P. E., Chiphatsi, S., Ismail, A., Kamau, T., Mwangi, I., English, M., Newton, C. R. J. C., Feikin, D. R., Scott, J. A. G. (2006). Effectiveness of Haemophilus influenzae type b Conjugate vaccine introduction into routine childhood immunization in Kenya.. JAMA 296: 671-678 [Abstract] [Full Text]  
• Tsang, R. S. W., Mubareka, S., Sill, M. L., Wylie, J., Skinner, S., Law, D. K. S. (2006). Invasive Haemophilus influenzae in Manitoba, Canada, in the Postvaccination Era. J. Clin. Microbiol. 44: 1530-1535 [Abstract] [Full Text]  
• Cerquetti, M., Cardines, R., Giufre, M., Castella, A., Rebora, M., Mastrantonio, P., Ciofi degli Atti, M. L. (2006). Detection of Six Copies of the Capsulation b Locus in a Haemophilus influenzae Type b Strain Isolated from a Splenectomized Patient with Fulminant Septic Shock. J. Clin. Microbiol. 44: 640-642 [Abstract] [Full Text]  
• Ho, P. L., Mak, G. C., Tse, C. W. S., Chow, K. H., Cheung, C. H. Y. (2006). Invasive Haemophilus influenzae isolates with decreased levofloxacin susceptibility in Hong Kong. J Antimicrob Chemother 57: 366-366 [Full Text]  
• Skoczynska, A., Kadlubowski, M., Empel, J., Hryniewicz, W. (2005). Characteristics of Haemophilus influenzae Type b Responsible for Meningitis in Poland from 1997 to 2004. J. Clin. Microbiol. 43: 5665-5669 [Abstract] [Full Text]  
• Sacchi, C. T., Alber, D., Dull, P., Mothershed, E. A., Whitney, A. M., Barnett, G. A., Popovic, T., Mayer, L. W. (2005). High Level of Sequence Diversity in the 16S rRNA Genes of Haemophilus influenzae Isolates Is Useful for Molecular Subtyping. J. Clin. Microbiol. 43: 3734-3742 [Abstract] [Full Text]  
• Scott, J. A. G., Mwarumba, S., Ngetsa, C., Njenga, S., Lowe, B. S., Slack, M. P. E., Berkley, J. A., Mwangi, I., Maitland, K., English, M., Marsh, K. (2005). Progressive Increase in Antimicrobial Resistance among Invasive Isolates of Haemophilus influenzae Obtained from Children Admitted to a Hospital in Kilifi, Kenya, from 1994 to 2002. Antimicrob. Agents Chemother. 49: 3021-3024 [Abstract] [Full Text]  
• Shen, K., Antalis, P., Gladitz, J., Sayeed, S., Ahmed, A., Yu, S., Hayes, J., Johnson, S., Dice, B., Dopico, R., Keefe, R., Janto, B., Chong, W., Goodwin, J., Wadowsky, R. M., Erdos, G., Post, J. C., Ehrlich, G. D., Hu, F. Z. (2005). Identification, Distribution, and Expression of Novel Genes in 10 Clinical Isolates of Nontypeable Haemophilus influenzae. Infect. Immun. 73: 3479-3491 [Abstract] [Full Text]  
• Sam, I.-C, Smith, M (2005). Failure to detect capsule gene bexA in Haemophilus influenzae types e and f by real-time PCR due to sequence variation within probe binding sites. J Med Microbiol 54: 453-455 [Abstract] [Full Text]  
• Nicolas, P., Djibo, S., Moussa, A., Tenebray, B., Boisier, P., Chanteau, S. (2005). Molecular Epidemiology of Meningococci Isolated in Niger in 2003 Shows Serogroup A Sequence Type (ST)-7 and Serogroup W135 ST-11 or ST-2881 Strains. J. Clin. Microbiol. 43: 1437-1438 [Abstract] [Full Text]  
• Skoczynska, A., Lewandowska, M., Klarowicz, A., Hryniewicz, W. (2005). Prevalence and Serotype Distribution of Encapsulated Haemophilus influenzae Isolates from Patients with Lower Respiratory Tract Infections in Poland. J. Clin. Microbiol. 43: 938-941 [Abstract] [Full Text]  
• Berkley, J. A., Lowe, B. S., Mwangi, I., Williams, T., Bauni, E., Mwarumba, S., Ngetsa, C., Slack, M. P.E., Njenga, S., Hart, C. A., Maitland, K., English, M., Marsh, K., Scott, J. A. G. (2005). Bacteremia among Children Admitted to a Rural Hospital in Kenya. NEJM 352: 39-47 [Abstract] [Full Text]  
• Westman, E., Lundin, S., Hermansson, A., Melhus, A. (2004). {beta}-Lactamase-Producing Nontypeable Haemophilus influenzae Fails To Protect Streptococcus pneumoniae from Amoxicillin during Experimental Acute Otitis Media. Antimicrob. Agents Chemother. 48: 3536-3542 [Abstract] [Full Text]  
• Sulikowska, A., Grzesiowski, P., Sadowy, E., Fiett, J., Hryniewicz, W. (2004). Characteristics of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis Isolated from the Nasopharynges of Asymptomatic Children and Molecular Analysis of S. pneumoniae and H. influenzae Strain Replacement in the Nasopharynx. J. Clin. Microbiol. 42: 3942-3949 [Abstract] [Full Text]  
• Watson, M. E. Jr, Burns, J. L., Smith, A. L. (2004). Hypermutable Haemophilus influenzae with mutations in mutS are found in cystic fibrosis sputum. Microbiology 150: 2947-2958 [Abstract] [Full Text]  
• Roman, F., Canton, R., Perez-Vazquez, M., Baquero, F., Campos, J. (2004). Dynamics of Long-Term Colonization of Respiratory Tract by Haemophilus influenzae in Cystic Fibrosis Patients Shows a Marked Increase in Hypermutable Strains. J. Clin. Microbiol. 42: 1450-1459 [Abstract] [Full Text]  
• Bajanca, P., Canica, M. (2004). Emergence of Nonencapsulated and Encapsulated Non-b-Type Invasive Haemophilus influenzae Isolates in Portugal (1989-2001). J. Clin. Microbiol. 42: 807-810 [Abstract] [Full Text]  
• Satola, S. W., Schirmer, P. L., Farley, M. M. (2003). Genetic Analysis of the Capsule Locus of Haemophilus influenzae Serotype f. Infect. Immun. 71: 7202-7207 [Abstract] [Full Text]  
• Bokermann, S., Zanella, R. C., Lemos, A. P. S., de Andrade, A. L. S. S., Brandileone, M. C. d. C. (2003). Evaluation of Methodology for Serotyping Invasive and Nasopharyngeal Isolates of Haemophilus influenzae in the Ongoing Surveillance in Brazil. J. Clin. Microbiol. 41: 5546-5550 [Abstract] [Full Text]  
• Campos, J., Roman, F., Perez-Vazquez, M., Aracil, B., Oteo, J., Cercenado, E., the Spanish Study Group for H. influenzae type f, (2003). Antibiotic resistance and clinical significance of Haemophilus influenzae type f. J Antimicrob Chemother 52: 961-966 [Abstract] [Full Text]  
• Augustynowicz, E., Gzyl, A., Szenborn, L., Banys, D., Gniadek, G., Slusarczyk, J. (2003). Comparison of usefulness of randomly amplified polymorphic DNA and amplified-fragment length polymorphism techniques in epidemiological studies on nasopharyngeal carriage of non-typable Haemophilus influenzae. J Med Microbiol 52: 1005-1014 [Abstract] [Full Text]  
• Bruant, G., Watt, S., Quentin, R., Rosenau, A. (2003). Typing of Nonencapsulated Haemophilus Strains by Repetitive-Element Sequence-Based PCR Using Intergenic Dyad Sequences. J. Clin. Microbiol. 41: 3473-3480 [Abstract] [Full Text]  
• Campos, J., Aracil, B., Roman, F., Perez-Vazquez, M. (2003). Molecular Epidemiology of Haemophilus influenzae Type b Isolated from Children with Clinical Cases of Conjugate Vaccine Failures. J. Clin. Microbiol. 41: 3915-3918 [Abstract] [Full Text]  
• Satola, S. W., Schirmer, P. L., Farley, M. M. (2003). Complete Sequence of the cap Locus of Haemophilus influenzae Serotype b and Nonencapsulated b Capsule-Negative Variants. Infect. Immun. 71: 3639-3644 [Abstract] [Full Text]  
• Anyanwu, J. N., Rodriguez, C. A., Fleming, K. E., Adderson, E. E. (2003). pgi Genotyping Is a Surrogate for Serotyping of Encapsulated Haemophilus influenzae. J. Clin. Microbiol. 41: 2080-2083 [Abstract] [Full Text]  
• Meats, E., Feil, E. J., Stringer, S., Cody, A. J., Goldstein, R., Kroll, J. S., Popovic, T., Spratt, B. G. (2003). Characterization of Encapsulated and Noncapsulated Haemophilus influenzae and Determination of Phylogenetic Relationships by Multilocus Sequence Typing. J. Clin. Microbiol. 41: 1623-1636 [Abstract] [Full Text]  
• Dabernat, H., Plisson-Saune, M.-A., Delmas, C., Seguy, M., Faucon, G., Pelissier, R., Carsenti, H., Pradier, C., Roussel-Delvallez, M., Leroy, J., Dupont, M.-J., De Bels, F., Dellamonica, P. (2003). Haemophilus influenzae Carriage in Children Attending French Day Care Centers: a Molecular Epidemiological Study. J. Clin. Microbiol. 41: 1664-1672 [Abstract] [Full Text]  
• Heath, P T, Booy, R, McVernon, J, Bowen-Morris, J, Griffiths, H, Slack, M P E, Moloney, A C, Ramsay, M E, Moxon, E R (2003). Hib vaccination in infants born prematurely. Arch. Dis. Child. 88: 206-210 [Abstract] [Full Text]  
• LaClaire, L. L., Tondella, M. L. C., Beall, D. S., Noble, C. A., Raghunathan, P. L., Rosenstein, N. E., Popovic, T. (2003). Identification of Haemophilus influenzae Serotypes by Standard Slide Agglutination Serotyping and PCR-Based Capsule Typing. J. Clin. Microbiol. 41: 393-396 [Abstract] [Full Text]  
• Cox, R A, Slack, M P E (2002). Clinical and microbiological features of Haemophilus influenzae vulvovaginitis in young girls. J. Clin. Pathol. 55: 961-964 [Abstract] [Full Text]  
• Taha, M.-K., Parent du Chatelet, I., Schlumberger, M., Sanou, I., Djibo, S., de Chabalier, F., Alonso, J.-M. (2002). Neisseria meningitidis Serogroups W135 and A Were Equally Prevalent among Meningitis Cases Occurring at the End of the 2001 Epidemics in Burkina Faso and Niger. J. Clin. Microbiol. 40: 1083-1084 [Abstract] [Full Text]  
• Williams, B. J., Morlin, G., Valentine, N., Smith, A. L. (2001). Serum Resistance in an Invasive, Nontypeable Haemophilus influenzae Strain. Infect. Immun. 69: 695-705 [Abstract] [Full Text]  
• Cerquetti, M., Ciofi degli Atti, M. L., Renna, G., Tozzi, A. E., Garlaschi, M. L., Mastrantonio, P., The Hi Study Group, (2000). Characterization of Non-Type B Haemophilus influenzae Strains Isolated from Patients with Invasive Disease. J. Clin. Microbiol. 38: 4649-4652 [Abstract] [Full Text]  
• Heath, P. T., Booy, R., Azzopardi, H. J., Slack, M. P. E., Bowen-Morris, J., Griffiths, H., Ramsay, M. E., Deeks, J. J., Moxon, E. R. (2000). Antibody Concentration and Clinical Protection After Hib Conjugate Vaccination in the United Kingdom. JAMA 284: 2334-2340 [Abstract] [Full Text]  
• Gomez-De-Leon, P., Santos, J. I., Caballero, J., Gomez, D., Espinosa, L. E., Moreno, I., Piñero, D., Cravioto, A. (2000). Genomic Variability of Haemophilus influenzae Isolated from Mexican Children Determined by Using Enterobacterial Repetitive Intergenic Consensus Sequences and PCR. J. Clin. Microbiol. 38: 2504-2511 [Abstract] [Full Text]  
• Leaves, N. I., Dimopoulou, I., Hayes, I., Kerridge, S., Falla, T., Secka, O., Adegbola, R. A., Slack, M. P. E., Peto, T. E. A., Crook, D. W. M. (2000). Epidemiological studies of large resistance plasmids in Haemophilus. J Antimicrob Chemother 45: 599-604 [Abstract] [Full Text]  
• Weller, T. M. A. (1999). The distribution of mecA, mecR1 and mecI and sequence analysis of mecI and the mec promoter region in staphylococci expressing resistance to methicillin. J Antimicrob Chemother 43: 15-22 [Abstract] [Full Text]  
• Smith-Vaughan, H. C., Sriprakash, K. S., Leach, A. J., Mathews, J. D., Kemp, D. J. (1998). Low Genetic Diversity of Haemophilus influenzae Type b Compared to Nonencapsulated H. influenzae in a Population in Which H. influenzae Is Highly Endemic. Infect. Immun. 66: 3403-3409 [Abstract] [Full Text]  
• Lo, T. M., Ward, C. K., Inzana, T. J. (1998). Detection and Identification of Actinobacillus pleuropneumoniae Serotype 5 by Multiplex PCR. J. Clin. Microbiol. 36: 1704-1710 [Abstract] [Full Text]  
• Heath, P T, Bowen-Morris, J, Griffiths, D, Griffiths, H, Crook, D W M, Moxon, E R (1997). Antibody persistence and Haemophilus influenzae type b carriage after infant immunisation with PRP-T. Arch. Dis. Child. 77: 488-492 [Abstract] [Full Text]