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Query: UMLS:C0348321 (Haemophilus)
 15,372 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

This study introduces a strategy to identify and produce sequences useful as genetic markers, or native genetic probes for DNA-DNA hybridization in bacterial strains where the genetics is not well described. Actinobacillus actinomy-cetemcomitans (A.a.) was used as an example. Fifty ng genomic DNA from A.a. ATCC 33384 and Haemophilus aphrophilus ATCC 33389 was amplified in a thermocycler using a single 10-mer primer. The PCR products were separated by electrophoresis on a 1% submarine agarose gel containing ethidium bromide and visualized by UV illumination, and the strain-specific amplitypes were compared. DNA from two bands, 0.9 and 4 kb, unique for the A.a. strain, was cut out, amplified under high stringency with the same primer and labeled by replacing 33.3 microM dTTP with digoxigenin-labeled dUTP in the reaction mixture. The labeled probe was then repeatedly used for hybridization to DNA from various A.a., H. aphrophilus, and other bacterial strains of the Pasteurellaceae family. The results showed that the 0.9-kb probe detected all A.a. tested, and distinguished it from other closely related bacterial species. We conclude that the described strategy is useful for identifying and selecting genetic sequences useful as genetic markers in A.a.
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PMID:Use of a nonradioactive genetic probe identified, synthesized, and labeled in the polymerase chain reaction. 752 67

The multiplex PCR method for the detection of Alloiococcus otitidis, Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae (P. H. Hendolin, A. Markkanen, J. Ylikoski, and J. J. Wahlfors, J. Clin. Microbiol. 35:2854-2858, 1997) in middle ear effusions (MEEs) was modified to be better suited for clinical use. To detect false-negative results, an internal amplification was added to the reaction, and to prevent carryover contamination, the dUTP-uracil-N-glycosidase system was incorporated into the procedure. Labor was minimized by using the heat-activatable AmpliTaq Gold polymerase in order to circumvent manual hot start and by detecting the amplification products on an automated sequencer. The performance of the improved protocol was verified with MEEs from patients with otitis media with effusion. In addition, a ligase detection reaction (LDR) was developed for confirmation of the PCR products. The modifications increased the reliability of the protocol and the hands-off time significantly. However, when two DNA extraction protocols were compared, gram-negative bacteria were detected more often in phenol-treated MEEs (94 versus 46%; P < 0.001), and gram-positive bacteria were detected more often in MEEs dissolved in sodium dodecyl sulfate-NaOH-chaotropic salt (83 versus 27%; P < 0.001). The LDR was found to be 100% specific. In all, the results demonstrate the feasibility of the rapid (7-h) multiplex PCR method for routine laboratory use.
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PMID:Clinically applicable multiplex PCR for four middle ear pathogens. 1061 75

Haemophilus somnus causes pneumonia, reproductive failure, infectious myocarditis, thrombotic meningoencephalitis, and other diseases in cattle. Although vasculitis is commonly seen as a result of systemic H. somnus infections, the pathogenesis of vascular damage is poorly characterized. In this study, we demonstrated that H. somnus (pathogenic isolates 649, 2336, and 8025 and asymptomatic carrier isolates 127P and 129Pt) induce apoptosis of bovine endothelial cells in a time- and dose-dependent manner, as determined by Hoechst 33342 staining, terminal deoxynucleotidyl transferase-mediated dUTP-FITC nick end labeling, DNA fragmentation, and transmission electron microscopy. H. somnus induced endothelial cell apoptosis in as little as 1 h of incubation and did not require extracellular growth of the bacteria. Viable H. somnus organisms induced greater endothelial cell apoptosis than heat-killed organisms. Since viable H. somnus cells release membrane fibrils and blebs, which contain lipooligosaccharide (LOS) and immunoglobulin binding proteins, we examined culture filtrates for their ability to induce endothelial cell apoptosis. Culture filtrates induced similar levels of endothelial cell apoptosis, as did viable H. somnus organisms. Heat inactivation of H. somnus culture filtrates partially reduced the apoptotic effect on endothelial cells, which suggested the presence of both heat-labile and heat-stable factors. We found that H. somnus LOS, which is heat stable, induced endothelial cell apoptosis in a time- and dose-dependent manner and was inhibited by the addition of polymyxin B. These data demonstrate that H. somnus and its LOS induce endothelial cell apoptosis, which may play a role in producing vasculitis in vivo.
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PMID:Haemophilus somnus induces apoptosis in bovine endothelial cells in vitro. 1117 40

Ribotyping profiles were determined for 12 Page serovar A isolates of Haemophilus paragallinarum from five outbreaks of infectious coryza in Hebei province in the People's Republic of China. Four enzymes were used to generate restriction digests of chromosomal DNA: HaeIII, HindIII, HpaII and SspI. The digests were probed with a digoxigenin-11-dUTP-labelled DNA probe produced by PCR amplification of the 16S rDNA of the type strain of H. paragallinarum. Only one profile was seen among the 12 isolates when using either HindIII or SspI. Three different ribotype profiles were seen with HpaII, while four different profiles were detected with HaeIII. Cluster analysis of the profiles generated by HpaII and HaeIII corresponded with the known epidemiological history of the isolates. These results are the first molecular characterization of isolates of H. paragallinarum from China and confirm the existence of genetic diversity in the H. paragallinarum population in that country.
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PMID:Molecular characterization of isolates of Haemophilus paragallinarum from China by ribotyping. 1848 95