The objective of this study was to develop a novel polymerase chain reaction (PCR) method to comprehensively analyze salivary bacterial flora.
The bacterial flora in the saliva of 10 healthy persons and 11 patients with odontogenic infections were examined using a DNA extraction method with a high level of cell destruction efficiency and a novel universal primer set to amplify approximately 580 bp of the 16S rDNA sequence.
Streptococcus (54.5%), Neisseria (14.7%), Actinomyces (8.4%), Gemella (4.1%), Granulicatella (3.8%), and Prevotella (1.4%) were dominant in a total of 1655 clones examined from the saliva of the healthy subjects. The dominant genera differed among the patients with odontogenic infections (a total of 823 clones) and were entirely different from those of the healthy subjects.
This novel comprehensive salivary bacterial flora analysis method may be a useful supportive method to identify causative agents of odontogenic infections.
[Show abstract][Hide abstract] ABSTRACT: Intracystic fluid was aseptically collected from 11 patients with postoperative maxillary cyst (POMC), and DNA was extracted from the POMC fluid. Bacterial species were identified by sequencing after cloning of approximately 580 bp of the 16S rRNA gene. Identification of pathogenic bacteria was also performed by culture methods. The phylogenetic identity was determined by sequencing 517-596 bp in each of the 1139 16S rRNA gene clones. A total of 1114 clones were classified while the remaining 25 clones were unclassified. A total of 103 bacterial species belonging to 42 genera were identified in POMC fluid samples by 16S rRNA gene analysis. Species of Prevotella (91%), Neisseria (73%), Fusobacterium (73%), Porphyromonas (73%), and Propionibacterium (73%) were found to be highly prevalent in all patients. Streptococcus mitis (64%), Fusobacterium nucleatum (55%), Propionibacterium acnes (55%), Staphylococcus capitis (55%), and Streptococcus salivarius (55%) were detected in more than 6 of the 11 patients. The results obtained by the culture method were different from those obtained by 16S rRNA gene analysis, but both approaches may be necessary for the identification of pathogens, especially of bacteria that are difficult to detect by culture methods, and the development of rational treatments for patients with POMC.
05/2012; 2012:840483. DOI:10.5402/2012/840483
"Until recently, the problem with developing the field of salivary diagnostics was that specific and informative biomarkers exist in saliva in relatively low quantities. However, the development of new, exquisitely sensitive amplification techniques such as RT-PCR, Q-PCR, and high-density oligonucleotide microarrays has demonstrated the feasibility of using saliva as a diagnostic probe for the rapid and unambiguous detection of oral biomarkers. Interestingly, saliva acts as a wide resource for genomic information useful for studying the potential disease status by analyzing their RNA level [Figure 1]. "
[Show abstract][Hide abstract] ABSTRACT: Saliva is a complex fluid, which influences oral health through specific and nonspecific physical and chemical properties. The importance of saliva in our everyday activities and the medicinal properties it possesses are often taken for granted. However, when disruptions in the quality or quantity of saliva do occur in an individual, it is likely that he or she will experience detrimental effects on oral and systemic health. Often head and neck radiotherapy has serious and detrimental side effects on the oral cavity including the loss of salivary gland function and a persistent complaint of a dry mouth (xerostomia). Thus, saliva has a myriad of beneficial functions that are essential to our well-being. Although saliva has been extensively investigated as a medium, few laboratories have studied saliva in the context of its role in maintaining oral and general health.
The Journal of Natural Science, Biology and Medicine 03/2011; 2(1):53-8. DOI:10.4103/0976-9668.82322
[Show abstract][Hide abstract] ABSTRACT: The aim of this study was to determine the influence of menstruation on the bacterial population of healthy Japanese women's vulvas, especially the labia minora. Labia minora swabs were obtained from 10 premenopausal, nonpregnant Japanese women at premenstruation and on day 2 of menstruation. Vaginal swabs were also obtained from 3 out of the 10 women. No significant difference was found in the average bacterial cell count between the menstruation and premenstruation samples. Molecular analysis using a 16S rRNA gene-based clone library method detected 22 genera from the labia minora swabs (total 20), with the genus Lactobacillus being predominant at both premenstruation and during menstruation in 7 out of the 10 women. Of the other 3 women, 2 showed various kinds of bacterial species, including oral and fecal bacteria, with Atopobium vaginae and Gardnerella vaginalis predominating in the remaining woman's vulva in both conditions. In total, 6 out of 10 cases (60%) showed significantly different microbiota of the labia minora between the two conditions. These results imply that menstruation may promote a distortion of the bacterial flora around the vulva, although it causes no significant increase of the bacterial count.
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