华西口腔医学杂志

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噻唑蓝比色法检测变异链球菌、血链球菌、伴放线菌嗜血菌的研究

王忠朝1 范丽苑2 蒋俊强1 蔡炜1 丁一3   

  1. 1.泸州医学院附属口腔医院口腔内科; 2.修复科, 四川泸州646000;3.四川大学华西口腔医院牙周病科, 四川成都610041
  • 收稿日期:2010-06-25 修回日期:2010-06-25 出版日期:2010-06-20 发布日期:2010-06-20
  • 通讯作者: 丁一,Tel:028-61153007
  • 作者简介:王忠朝(1980—),男,四川人,住院医师,硕士

Study on the counting of Streptococcus mutans, Streptococcus sanguis, Haemophilus actinomycetemcomitans by methyl thiazolyl tetrazolium colorimetric method

WANG Zhong-chao1, FAN Li-yuan2, JIANG Jun-qiang1, CAI Wei1, DING Yi3   

  1. 1. Dept. of Endodontics, Stomatological Hospital of Luzhou Medical College, Luzhou 646000, China; 2. Dept. of Prosthodontics, Stomatological Hospital of Luzhou Medical College, Luzhou 646000, China; 3. Dept. of Periodontology, West China College of Stomatology, Sichuan University, Chengdu 610041, China
  • Received:2010-06-25 Revised:2010-06-25 Online:2010-06-20 Published:2010-06-20
  • Contact: DING Yi,Tel:028-61153007

摘要:

目的探讨噻唑蓝(MTT)法用于口腔常见细菌(变异链球菌、血链球菌、伴放线菌嗜血菌)活菌计数的可行性及具体应用过程中的实验条件。方法本实验以菌落形成单位(CFU)法为标准对照。实验通过改变比色的波长、反应时间、试剂剂量、细菌菌龄等实验条件,获得MTT法测量常见口腔细菌的一系列参数,对变异链球菌、血链球菌、伴放线菌嗜血菌进行活菌计数,并与CFU法所测的细菌数量相比较。结果MTT法在测量变异链球菌时,最佳的检测波长为510 nm,最佳的测定细菌范围是1.5×105~1.0×107 CFU·mL-1。MTT法在测量血链球菌时,最佳的检测波长为545 nm,最佳的测定细菌范围是1.5×105~2.0×107 CFU·mL-1。MTT法在测量伴放线菌嗜血菌时,最佳的检测波长为557 nm,最佳的测定细菌范围是1.0×106~5.0×107 CFU·mL-1。MTT法与菌落形成单位法的测量结果是一致的,并且对于不同菌龄的变异链球菌、血链球菌、伴放线菌嗜血菌均适用。结论MTT法可以用于检测变异链球菌、血链球菌、伴放线菌嗜血菌等口腔常见细菌的活菌数量,并且具有快速、方便等优点。

关键词: 噻唑蓝比色法, 变异链球菌, 血链球菌, 伴放线菌嗜血菌, 活菌计数

Abstract:

Objective To explore the feasibility of methyl thiazolyl tetrazolium(MTT) colorimetric method and the applied condition for the normal bacteria in the mouth, as Streptococcus mutans(S.mutans), Streptococcus sanguis(S. sanguis), Haemophilus actinomycetemcomitans(H.actinomycetemcomitans). Methods Colony forming units(CFU) which was the standard antitheses was used to count bacteria. This study would gain some parameters by changing wavelength, reactive time, dosage and so on. MTT colorimetric method was applied in the counting of S.mutans, S. sanguis and H.actinomycetemcomitans. Results When counting S.mutans, the best wavelength was 510 nm, the best range was 1.5×105~1.0×107 CFU·mL-1. When counting S.sanguis, the best wavelength was 545 nm, the best range was 1.5×105~2.0 ×107 CFU·mL -1. When counting H.actinomycetemcomitans, the best wavelength was 557 nm, the best range was 1.0×106~5.0×107 CFU·mL-1. MTT colorimetric method can be used for different aged S.mutans, S.sanguis and H.actinomycetemcomitans. Conclusion Oral bacteria could be counted by MTT colorimetric method, which is fast and convenient.

Key words: methyl thiazolyl tetrazolium colorimetric method, Streptococcus mutans, Streptococcus sanguis, Haemophilus actinomycetemcomitans, bacterial cell counting