Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1990-08-29
1993-05-18
Wax, Robert A.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 42, 536 2432, C12G 168, C07H 1512, C12P 3900
Patent
active
052120597
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
Technical Field of the Invention
This invention relates to compositions of oligonucleotide probes for use in the detection of bacteria associated with medical disorders of the human mouth, wherein said probes consist essentially of a segment of nucleic acid capable of selectively hybridizing under hybridizing conditions, to the 16S or 23S ribosomal RNA [rRNA] of said bacteria. Methods for detection, as well as diagnostic kits for the assay of these bacteria, are also disclosed.
Substantial evidence exists implicating plaque bacteria in the etiology of human periodontal disease, particularly gingivitis and periodontitis. As these diseases have been studied, increasingly sophisticated methods for detection, quantitation, and identification of specific oral bacteria and other infectious viruses and microorganisms have been developed. These improved methods have led to the identification of specific pathogenic bacteria playing a major role in the etiology of periodontal disease.
Human periodontal diseases constitute a major health-care problem in the United States. While epidemiologic surveys suggest that there may be a reduction of the proportion of Americans with gingivitis in recent years, the number of patients having some stage of the disease remains high. Furthermore, the proportion of adults, aged 18-79 years, with periodontal pockets appears to be relatively unchanged.
In the early 1970's, roughly 30 percent of Americans had significant periodontal disease problems by age 50, and by age 60, between 30 and 40 percent of all Americans had lost all of their teeth, primarily due to periodontal disease. At present, the exact proportion of persons with severe periodontal disease is unknown, but recent estimates are that 10-20 percent of the overall American population has periodontal disease serious enough to result in tooth loss. Coincidental with the disease is a significant relationship between the onset and course of the disease, and identifiable (suspect) pathogenic bacterial species.
Although the precise complex mechanisms involved in the pathogenesis of human periodontitis are still unclear, there is wide agreement that bacteria that colonize the gingival crevice area around the teeth are of primary etiologic importance. Within the past decade, researchers at several academic institutions have demonstrated that of the large number of bacterial genera and species found in the human periodontal crevice or pocket (ca. 256 species), a relatively limited number of these appear to be consistently associated with periodontal disease. These microorganisms have been designated "suspect periodontal pathogens." Of particular interest are the microorganisms shown in Table 1.
INFORMATION DISCLOSURE
The detection and characterization of the periodontal pathogenic bacteria present in a given periodontal pocket site are, by current methods, both time-consuming and expensive (e.g., microscopic, cultural, gas chromatographic and metabolic product analysis). Furthermore, highly trained individuals, with sophisticated laboratory equipment, are required. Most of the published studies on the microbiology of human periodontal diseases have been compromised by the enormous workload that must be accomplished to isolate and characterize individual members of the microflora at each periodontal site. As a result, the number of sites sampled per patient and the number of patients examined are limited. Clearly, the present methods do not allow routine testing of periodontal patients for the presence of pathogenic bacteria.
Immunoassays are reported as an alternative to the above detection methodologies. Fluorescent monoclonal antibodies to Actinobacillus actinomycetemcomitans (A.a.), Bacteroides gingivalis (B.g.), and Bacteroides intermedius (B.i.) have been developed. Chen, P., et al., The Use Of Monoclonal Antibodies To Detect Bacteroides gingivalis In Biological Samples, Infection and Immunity, 54:798-803 (1986); Gmur, R., et al., Quantitative Monitoring Of The Periodontal Pocket Flora With Mo
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Dix Kim
Kanemoto Roy H.
Schwartz Dennis E.
Watanabe Susan M.
Escallon M.
Microprobe Corporation
Wax Robert A.
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