An analysis of the diversity ofHaemophilus parainfluenzaein the adult human respiratory tract by genomic DNA fingerprinting

Abstract

SUMMARYA method for typing Haemophilus species is described, based on the analysis of genomic DNA from Haemophilus parainfluenzae. The DNA was extracted by a rapid method and digested with the restriction enzyme BamHI to provide a characteristic 'fingerprint'. The pattern of fragments in the ranges 1-1 6 kb, 1-6-2 kb and 2-3 kb were used to produce a numerical profile of each isolate. In total 97 isolates were examined; 88 from throat swab material isolated from the 15 members of a British Antarctic Survey base and 9 type strains. Seventy-two of the 88 antarctic isolates were H. parainfluenzae and were found to be very diverse, comprising 41 identifiable strains with up to 5 strains being isolated from a single throat swab sample. There was evidence for both carriage and transmission within the isolated community. The technique provided a highly discriminatory method for characterizing Haemophilus strains which is suitable for epidemiological studies.

Population Dynamics of Streptococcus mitis in Its Natural Habitat

“…The results furthermore confirm that the S. mitis biovar 1 population, even in well-defined, narrow, and homogeneous habitats like the buccal and oropharyngeal mucosae, consists of a multitude of distinct genotypes (15,22). Degrees of clonal diversity within single individuals approaching that of S. mitis biovar 1 have been demonstrated for Eikenella corrodens, Actinomyces naeslundii, and Haemophilus parainfluenzae (4,13,23).…”

“…In contrast, genetic typing of bacteria that are commensals or opportunistic pathogens have revealed considerably more diversity, sometimes even within a single host (1,2,7,11,13,15,22,23,37,49), although typing methods such as restriction endonuclease analysis (REA), ribotyping, or other highly sensitive DNA-based typing methods may miss the clonal relationships of types. However, only rarely are the causes and ecological significance of this diversity considered.…”

Molecular characterization of a pre‐Columbian mummy and in situ coprolite

“…Unfortunately, only very recently has the phylogeny of the genus Haemophylus been the object of attention by specialists (Hedegaard et al, 2001;Nørskov-Lauritsen et al, 2005).These works, however, are mainly aimed at verifying the correspondence between traditional and molecular methods of bacterial classification, rather than reconstructing an evolutionary history of the pathogens. In addition, the techniques used in the recent past to differentiate among biotypes of H. parainfluenzae are either based on amino-acid metabolism (Taylor et al, 1992) or DNA fingerprinting based on the analysis of restriction-fragment patterns in the range of 1-3 kb (Kerr et al, 1993), and thus are totally unsuitable for the analysis of archaeological specimens. Other methods such as PCR-ribotyping (Privitera et al, 1998), based on identification of length polymorphisms in the intergenic spacer regions of Haemophilus spp rDNA, were only able to distinguish between H. influenzae and H. parainfluenzae, but not to discriminate within species.…”

Incidence and spread ofHaemophilus influenzaeon an Antarctic base determined using the polymerase chain reaction

“…The most recent of such studies was that of Kerr and colleagues (4), who investigated the carriage and transmission of Haemophilus parainfluenzae in the inhabitants of Signy base in the South Orkney Islands. In this paper we report the results of a study which used the polymerase chain reaction (PCR) and DNA restriction enzyme analysis to investigate the carriage and spread of Haemophilus influenzae in contemporaneous samples from the subjects of Kerr and colleagues [4].…”

“…Each of the samples from the microbiologists was plated out onto two agar plates, one of which was 'spiked ' by the addition to its inoculum of a loopful of Hib bacteria from a pure culture before spreading and incubation. Antarctic samples were the duplicate freeze-dried samples of Kerr and colleagues [4], which had been obtained from the left tonsillar fossae of the Antarctic subjects during the first week of each month between February and September 1990 and cultured on non-selective media prior to freeze drying. Freeze-dried material was rehydrated with 200 #1 sterile distilled water and inoculated on vancomycin supplemented chocolate agar plates for 24 h culture.…”