Two nested PCRs for the detection of Mycobacterium ulcerans were compared by using a collection of 65 clinical specimens. The first method amplifies the gene coding for 16S rRNA, and the second method amplifies a repetitive DNA sequence. The sensitivities of bacterioscopy, culture, 16S rRNA gene PCR, and repetitive-sequence PCR were 29, 34, 80, and 85%, respectively. Compared to the 16S rRNA ge...

BACKGROUND The intra- and inter-species genetic diversity of bacteria and the absence of 'reference', or the most representative, sequences of individual species present a significant challenge for sequence-based identification. The aims of this study were to determine the utility, and compare the performance of several clustering and classification algorithms to identify the species of 364 seq...

Pseudomonas aeruginosa is a gram negative bacterium isolated from soil. The isolate was cultured in a selective medium (Pseudomonas Agar base) at 37oC for 24 hours. For species identification, Pseudomonas like organisms creates lot of problems when identified with the help of morphological and biochemical characters. However, sequencing of 16S rRNA region is a suitable technique for species ide...

The RDP Classifier is a widely used bioinformatic program that performs taxonomic classification of 16S rRNA gene sequences. However, the accuracy of the program is not clear when it is applied to common PCR products of the 16S rRNA variable regions, which are heavily used in microbiome projects. In this study, fulllength 16S rRNA gene alignments from the SILVA database were used to simulate th...

16S rRNA gene sequencing has been widely used for probing the species structure of a variety of environmental bacterial communities. Alternatively, 16S rRNA gene fragments can be retrieved from shotgun metagenomic sequences (metagenomes) and used for species profiling. Both approaches have their limitations-16S rRNA sequencing may be biased because of unequal amplification of species' 16S rRNA ...

The traditional identification of bacteria on the basis of phenotypic characteristics is generally not as accurate as identification based on genotypic methods. Comparison of the bacterial 16S rRNA gene sequence has emerged as a preferred genetic technique. 16S rRNA gene sequence analysis can better identify poorly described, rarely isolated, or phenotypically aberrant strains, can be routinely...

Even though the 16S rRNA gene is the most commonly used taxonomic marker in microbial ecology, its poor resolution is still not fully understood at the intra-genus level. In this work, the number of rRNA gene operons, intra-genomic heterogeneities and lateral transfers were investigated at a fine-scale resolution, throughout the Pseudomonas genus. In addition to nineteen sequenced Pseudomonas s...

Although the applicability of small subunit ribosomal RNA (16S rRNA) sequences for bacterial classification is now well accepted, the general use of these molecules has been hindered by the technical difficulty of obtaining their sequences. A protocol is described for rapidly generating large blocks of 16S rRNA sequence data without isolation of the 16S rRNA or cloning of its gene. The 16S rRNA...

background aggregatibacter actinomycetemcomitans and tannerella forsythensis are two major pathogens in destructive periodontal disease in humans. the detection of these bacteria is needed for diagnosis and management of the mentioned diseases. objectives we aimed to develop and compare improved multiplex conventional and sybr green real time pcr assays for a specific diagnosis of the organisms...