| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2002: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2001: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2000: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Research Abstract |
Nutritionally variant streptococci (NVS) are isolated from normal oral flora and various infections disease sites. The NVS are classified into four species (Abiotrophia defectiva, Granulicatella adiacens, Granulicatella paradiacens, Granulicatella elegans). Characteristically, vitamin B6 analogue is necessary to grow of NVS. This necessity limits detection and identification of the NVS in infectious disease sites. This limitation often brings "culture negative" in clinical diagnosis.
Detection and identification of pathogenic species by PCR method is efficacious against culture-negative organism. Primers based on 16S rRNA sequences are used for the PCR. 16S rRNA gene is conserved and the sequence resembles among species. If a single species exist in a disease site, the primers derived from 16S rRNA sequences are effective. However, in many cases, pathogenic species resides in multiple bacterial infections. For those cases, species-specific primers are needed.
We found species-specific primers for NVS species. Primers AD, GA, GP, and GE are exactly specific for A. defectiva, G. adiacens, G. paradiacens, and G. elegans, respectively. These primers are effective for detection and identification of NVS in clinical specimens obtained from human dental plaques. These results suggest that these primers should be useful for the clinical diagnosis of infectious diseases.
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