| Objectives: Decayed tooth sickness is one kind of severely impair humanity health most common oral disease, the milk teeth decayed tooth is in child oral cavity's common disease and frequently-occurring disease, this research in the earlier period experiment's foundation, carries on the discussion to different decayed tooth sensitive child oral cavity variation chain coccus clinical separation HtrA mRNA and the protein expression situation, the analysis sends the decayed tooth strain HtrA expression differently the difference, obtains the milk teeth decayed tooth bad degree and the HtrA relations, to consummate the decayed tooth risk forecast technique to provide the powerful theory and the experiment rests on. Methods: 1. Obtains by the earlier period experiment has different gene nuclear type Gao Qu, the decayed tooth loses makes up the index (decayed, missing, filled teeth index, dmft)≥6, the decayed tooth, 6>dmft≥4, does not have the decayed tooth, in the dmft=0 altogether 111 variation chain coccuses selects 45 (Gao Quzu 15, decayed tooth group 15, do not have decayed tooth group 15) is the experiment strains, take the international standard strain distortion chain coccus (S.mutans) ATCC 25175 (blood serum C) as the masculine comparison, Yu Qing the saliva agar-agar (MS) the plate recovers, smear inspection after pure raise, takes the typical colony vaccination to dip (Brain in the ox heart brain Heart Infusion, BHI), 37°C anaerobic (80%N2, 10%H2, 10%CO2) raise 24h, the separation purification nucleic acid, uses counter copies PCR (reverse transcription PCR, RT-PCR) the law, extraction distortion chain coccus clinical separation total RNA, examines the RNA integrity with 1% agarose gel electrophoresis, synthesizes cDNA and takes the cDNA product to further carry on PCR to increase, responded after having finished, will increase the product to carry on 2.0% agarose gel electrophoresis, take DL500 DNA Marker as the molecular weight reference standard, under the gelatin imaging system observes the record result, obtained goal gene HtrA and the materials for internal reference electrophoresis image 4.0 will carry on the gradation scanning using gelatin quantitative analysis software Gel-Pro analyzer, the analysis computation gene relative expression value. 2. The separation purification protein, uses the protein signature (Western Blot) the law, an extraction distortion chain coccus clinical separation total protein, examines the total protein sample density, the polyacrylamide gelatin (SDS-PAGE) the ionophortic separation, after testing the Mas light blue dyes, transfers the membrane, joins 5% nonfat dry milk, after the room temperature seals up 1h hybridizes, the chemiluminescence development, the result scans into the computer with the Bio-Rad gelatin photograph analysis system, gelatin quantitative analysis software Quantity One 4.4.0 analyze its grey level, the expression protein relative expression level. Results: 1. Gao Qu, the decayed tooth and non-decayed tooth child oral cavity variation chain coccus clinical separation HtrA the mRNA expression existence difference, Gao Quzu > the decayed tooth group > does not have the decayed tooth group, the difference has statistics significance (p<0.05). 2. Gao Qu, the decayed tooth and the non-decayed tooth child oral cavity variation chain coccus clinical separation HtrA protein's expression also has the difference, Gao Quzu > the decayed tooth group > does not have the decayed tooth group, the difference has statistics significance (p<0.05). Conclusion: 1. different decayed tooth sensitive child oral cavity variation chain coccus clinical separation strain HtrA the mRNA expression existence difference, the decayed tooth sensitivity is higher, HtrA the mRNA expression is higher. 2. The different decayed tooth sensitive child oral cavity variation chain coccus clinical separation strain HtrA protein expression existence difference, the decayed tooth sensitivity is higher, the HtrA protein expression is higher.
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