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Bacteriological Methods (bacteriological + methods)
Selected AbstractsBacteria of asymptomatic periradicular endodontic lesions identified by DNA-DNA hybridizationDENTAL TRAUMATOLOGY, Issue 5 2000J. J. Gatti Abstract , Possible inclusion of contaminant bacteria during surgery has been problematic in studies of periradicular lesions of endodontic origin. Therefore, in this study, two different surgical techniques were compared. A second problem is that some difficult to cultivate species may not be detected using bacteriological methods. Molecular techniques may resolve this problem. DNA-DNA hybridization technology has the additional advantage that DNA is not amplified. The purpose of this investigation was to determine if bacteria from periradicular endodontic lesions could be identified using DNA-DNA hybridization. A full thickness intrasulcular mucoperiosteal (IS) flap (n=20) or a submarginal (SM) flap (n=16) was reflected in patients with asymptomatic apical periodontitis. DNA was extracted and incubated with 40 digoxigenin-labeled whole genomic probes. Bacterial DNA was detected in all 36 lesions. Seven probes were negative for all lesions. In patients with sinus tract communication, in teeth lacking intact full coverage crowns, and in patients with a history of trauma, 4,13 probes provided positive signals. Seven probes were positive in lesions obtained by the IS, but not the SM technique. Two probes were in samples obtained with the SM technique, but not the IS. Only Bacteroides forsythus and Actinomyces naeslundii genospecies 2 were present in large numbers using either the IS or the SM technique. The SM flap technique, in combination with DNA-DNA hybridization, appeared to provide excellent data pertaining to periradicular bacteria. These results supported other studies that provide evidence of a bacterial presence and persistence in periradicular lesions. [source] Subgingival microbial profiles in chronic periodontitis patients from Chile, Colombia and SpainJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 2 2008David Herrera Abstract Aim: To investigate the subgingival microbiota of distinct periodontitis patient populations, in Chile, Colombia and Spain, using identical clinical and bacteriological methods. Material and Methods: In this multicentre study, 114 chronic periodontitis patients were selected. Patients were examined using an identical clinical protocol and pooled subgingival samples were obtained from each patient. Samples were processed in the three laboratories by means of culturing under identical clinical and microbiological protocols. Total anaerobic counts and frequency of detection and proportions of nine periodontal pathogens were calculated. Variables were analysed by means of anova, ,2, Kruskal,Wallis and Dunn's multiple comparison tests. Results: The Colombian population demonstrated greater severity of periodontitis, with significantly deeper mean probing pocket depth, and had a significantly lower percentage of current smokers. When comparing samples from the three patient populations, the total counts were significantly higher in the Colombian patients. The numbers of putative pathogens differed among groups. Tannerella forsythia was found less frequently in Chilean samples, while Parvimonas micra and enteric rods differed significantly among the three population groups. Conclusion: Significant differences among Chile, Colombia and Spain existed regarding the frequency and proportions of specific periodontal pathogens in the subgingival microbiota of periodontitis patients. [source] Humoral immune response against 38- and 16-kDa mycobacterial antigens in childhood tuberculosis,PEDIATRIC PULMONOLOGY, Issue 9 2009Gunes Senol MD Abstract Several enzyme-linked immunosorbent assays (ELISAs) based on mycobacterial antigens have been tried for the rapid diagnosis of tuberculosis (TB). In this study, the value of the 16 and 38-kDa mycobacterial antigens in the diagnosis of TB was investigated in pediatric patients in Izmir, Turkey in whom they were found using clinical and/or bacteriological methods. A commercial ELISA kit was used for measuring IgG against 38 and 16-kDa recombinant antigens. The humoral immune response was analyzed in a group of 32 TB patients (24 pulmonary, 3 lymphadenitis and 2 pleuritis, 2 meningitis and a disseminated TB) and in control groups consisting of 20 healthy children and 20 pulmonary diseases other than TB cases. The sensitivity, specificity, positive predictive value, and the negative predictive value of the test were found to be 25%, 90%, 66.7%, and 60%, respectively, in the TB cases. The ELISA test shows very good specificity, but low level of sensitivity and negative predict value. It was thought that it might be used in combination with other methods to increase diagnostic accuracy, especially for culture-negative TB pediatric cases, which are difficult to diagnose. Pediatr Pulmonol. 2009; 44:839,844. © 2009 Wiley-Liss, Inc. [source] Multiplex nested-polymerase chain reaction for the simultaneous detection of Aeromonas hydrophila, Edwardsiella tarda, Photobacterium damselae and Streptococcus iniae, four important fish pathogens in subtropical AsiaAQUACULTURE RESEARCH, Issue 10 2009Chin-I Chang Abstract A multiplex nested-polymerase chain reaction (PCR)-based (m-nested PCR) method was developed for simultaneous detection of four important freshwater/marine fish pathogens in subtropical Asia, including Aeromonas hydrophila, Edwardsiella tarda, Photobacterium damselae and Streptococcus iniae. The specificity of the oligonucleotide primers used for PCR detection was confirmed to generate specific amplicons for the corresponding pathogens. Moreover, non-specific amplicons were observed when the primers were tested using pure DNA extracted from 31 related bacterial strains belonging to 23 species or tissue homogenates of infected tilapia. This m-nested PCR approach could detect 19 colony forming unit (CFU) for A. hydrophila, 62 CFU for E. tarda, 280 CFU for P. damselae subsp. piscicida and 179 CFU for S. iniae in infected tilapia kidney homogenates, consistent with the results derived from bacteriological methods. The assay described in this paper is a sensitive and effective method for simultaneous detection of multiple fish pathogens. [source] |