Инфекция и иммунитет. 2020; 10: 747-754
Роль инфекционного агента в развитии поражения зубов
https://doi.org/10.15789/2220-7619-TRO-1138Аннотация
Цель: оценить влияние колонизации полости рта штаммами S. mutans с различными генотипическими характеристиками на степень пораженности зубов у детей.
Материалы и методы. В исследование были включены 274 ребенка в возрасте от 5 до 17 лет (153 девочки и 121 мальчик), которым проводился профилактический стоматологический осмотр. Степень пораженности зубов оценивалась по индексу КПУ (сумма кариозных, пломбированных, удаленных по причине кариеса зубов), согласно рекомендациям ВОЗ. Зубной налет собирался стерильными деревянными зубочистками с щечно-десневого края либо с фиссур первых моляров и помещался в 1,5 мл пробирки Eppendorf, затем проводился посев на среду Mitis Salivarius Agar (HiMedia, REF, Индия). Для дальнейшего изучения был отобран 481 штамм S. mutans. Выделение ДНК осуществлялось экспресс-методом. Амплификация выполнялась в термоциклере CFX-96 (Bio-Rad, США). Серотипирование проводилось методом мультиплекс-ПЦР. Анализ ПЦР-продуктов осуществлялся методом гель-электрофореза в 1,5% агарозном геле с окраской ДНК этидиумом бромида (10 мг/мл) производства Helicon (Москва) и визуализацией в УФ-лучах на трансиллюминаторе UVT1 производства Biokom. Генотипи-рование осуществлялось согласно методике (Saarela и соавт., 1996) олигонуклеотидным праймером OPA-02 (5'-TGCCGAGCTG-3'). Штаммы S. mutans были изучены на наличие следующих генов: gtfB, spaP, cnm, fruA, gtfB, htrA, comE, mutA (I), mutA (II), mutA (III), nlmAB (IV), adcA, Smu.399, Smu.583, Smu.761, Smu.940c, Smu.1449, Smu.2130.
Результаты. S. mutans был выделен у всех обследованных детей. Поражение зубов выявлено у 82,4% детей. Среди изучаемых штаммов были обнаружены все 4 серотипа: у детей с индексом КПУ = 0 обнаруживались только серотипы k и f, доминирующим серотипом у детей с поражением зубов являлся с (74,7%). Были идентифицированы 19 генотипов S. mutans. У детей без кариеса (индекс КПУ = 0) S. mutans не содержали гены spaP, comE, adcA, Smu.2130, smu1449, gtfB, htrA. С увеличением индекса КПУ увеличивалась частота их обнаружения. Все 7 факторов вирулентности имели 9 генотипов S. mutans. При колонизации данными «вирулентными» генотипами у 94,9% детей отмечался высокий индекс КПУ.
Заключение. Полученные данные свидетельствуют о том, что лишь отдельные штаммы обладают кариесогенным потенциалом. У детей с поражением зубов были выделены штаммы S. mutans серотипов е и с, имеющие сочетание генов вирулентности: spaP, gtfB, comE, adcA, Smu.2130, Smu.1449, htrA. Штаммы, не имеющие данных факторов, повреждений зубов не вызывали. Степень пораженности зубов увеличивается при колонизации несколькими генотипами с данным профилем факторов вирулентности.
Список литературы
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14. Palmer E.A., Vo A., Hiles S.B., Peirano P., Chaudhry S., Trevor A., Kasimi I., Pollard J., Kyles C., Leo M., Wilmot B., Engle J., Peterson J., Maier T., Machida C.A. Mutans streptococci genetic strains in children with severe early childhood caries: follow-up study at one-year post-dental rehabilitation therapy. J. Oral Microbiol., 2012, no. 4, pp. 167-173. doi: 10.3402/jom.v4i0.19530
15. Rincon-Rodriguez R.J., Parada-Sanchez M.T., Bedoya-Correa C.M., Arboleda-Toro D. Genetic diversity of Streptococcus mutans serotype c isolated from white spot and cavitated caries lesions from schoolchildren. Arch. Oral Biol, 2019, vol. 100, pp. 33-41. doi: 10.1016/j.archoralbio.2019.02.003
16. Struzycka I. The oral microbiome in dental caries. Pol. J. Microbiol., 2014, vol. 63, no. 2, pp. 127-135.
17. Suzuki S., Ukiya T., Kawauchi Y., Ishii H., Sugihara N. Decision tree analysis for factors associated with dental caries in school-aged children in Japan. Community Dent. Health., 2018, vol. 35, no. 4, pp. 247-251. doi: 10.1922/CDH_4409Suz.uki05
18. Zhou Q., Qin X., Qin M., Ge L. Genotypic diversity of Streptococcus mutans and Streptococcus sobrinus in 3-4-year-old children with severe caries or without caries. Int. J. Paediatr. Dent., 2011, vol. 21, no. 6, pp. 422-431. doi: 10.1111/j.1365-263X.2011.01145.x
Russian Journal of Infection and Immunity. 2020; 10: 747-754
The role of infectious agent in development of tooth decay
https://doi.org/10.15789/2220-7619-TRO-1138Abstract
Aim: to assess the relationship between colonization of the oral cavity with S. mutans and different genotypic characteristics and the degree of tooth decay in children.
Materials and methods. 274 children aged 5 to 17 years (153 girls and 121 boys) who received a preventive dental checkup were included in the study. The dental caries experience was assessed by the DMFT index (number of decayed, missing due to caries, and filled teeth), according to WHO recommendations. The plaque was collected with sterile wooden toothpicks from the buccal gingival margin or from fissures of the first molars and placed in 1.5 mL Eppendorf tubes, and then plated on Mitis Salivarius Agar medium (HiMedia, India). 481 strains of S. mutans were selected for further study. DNA was extracted by an express method. Amplification was performed in the CFX-96 thermal cycler (Bio-Rad, USA). Serotyping was performed by multiplex PCR. PCR products were analyzed by gel electrophoresis in 1.5% agarose gel with ethidium bromide (10 mg/mL) manufactured by Helicon, Moscow, and visualized in UV light in transilluminator UVT1 by Biokom. Genotyping was performed according to the methodology (Saarela et al., 1996) with the oligonucleotide primer OPA-02 (5’-TGCCGAGCTG-3’). Strains of S. mutans were studied for the presence of the following genes: gtfB, spaP, cnm, fruA, gtfB, htrA, comE, mutA x(I), mutA (II), mutA (III), nlmAB (IV), adcA, Smu.399, Smu.583, Smu.761, Smu.940c, Smu.1449, Smu.2130.
Results. S. mutans was isolated from all the examined children. Dental decay was detected in 82.4% of the children. Among the strains studied, all 4 serotypes were found: in children with a DMFT = 0 only serotypes k and f were detected; the predominant serotype in children with tooth decay was serotype c (74.7%). 19 genotypes of S. mutans were identified. In children without caries (DMFT = 0), S. mutans did not contain the genes spaP, comE, adcA, Smu.2130, Smu.1449, gtfB, htrA. With the increase in the DMFT index, the frequency of their detection increased. 9 genotypes of S. mutans had all 7 virulence factors. In 94.9% of children colonized by these “virulent” genotypes, high DMFT index scores were observed.
Conclusion. The data obtained indicate that only a limited number of specific strains have a cariogenic potential. Strains of S. mutans belonging to serotypes e and c with a combination of virulence genes spaP, gtfB, comE, adcA, Smu.2130, Smu.1449, and htrA were isolated from children with tooth decay. Strains without these factors did not cause any damage to the teeth. The degree of tooth decay increases with colonization by several genotypes with the combination of virulence factors described above.
References
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3. Bottner A., He R.Y., Sarbu A., Nainar S.M.H., Dufour D., Gong S.G., Levesque C.M. Streptococcus mutans isolated from children with severe-early childhood caries form higher levels of persisters. Arch. Oral Biol., 2020, vol. 110: 104601. doi: 10.1016/j.archoralbio.2019.104601
4. Carletto-Korber F.P., Gonzalez-Ittig R.E., Jimenez M.G., Cornejo L.S. Serotype diversity of Streptococcus mutans and caries activity in children in Argentina. Eur. J. Paediatr. Dent., 2015, vol. 16, no. 3, pp. 177—180.
5. Cheon K., Moser S.A., Wiener H.W., Whiddon J., Momeni S.S., Ruby J.D., Cutter G.R., Childers N.K. Characteristics of Streptococcus mutans genotypes and dental caries in children. Eur. J. Oral Sci., 2013, vol. 121, no. 3, pt. 1, pp. 148—155. doi: 10.1111/eos.12044
6. Dame-Teixeira N., Arthur R.A., Parolo C.C., Maltz M. Genotypic diversity and virulence traits of Streptococcus mutans isolated from carious dentin after partial caries removal and sealing. ScientificWorldJournal, 2014, vol. 21:165201. doi: 10.1155/2014/165201
7. Fan C.C., Wang W.H., Xu T., Zheng S.G. Risk factors of early childhood caries (ECC) among children in Beijing — a prospective cohort study. BMC Oral Health, 2019, vol. 19, no. 1, p. 34. doi: 10.1186/s12903-019-0721-9
8. Gomes M.C., Pinto-Sarmento T.C., Costa E.M., Martins C.C., Granville-Garcia A.F., Paiva S.M. Impact of oral health conditions on the quality of life of preschool children and their families: a cross-sectional study. Health Qual. Life Outcomes, 2014, no. 12, pp. 55- 63. doi: 10.1186/1477-7525-12-55
9. Kreth J., Merritt J., Zhu L., Shi W., Qi F. Cell density- and ComE-dependent expression of a group of mutacins and mutacin-like gene in Streptococcus mutans. FEMSMicrobiol. Lett., 2006, vol. 265, no. 1, pp. 11-17. doi: 10.1111/j.1574-6968.2006.00459.x
10. Lembo F.L., Longo P.L., Ota-Tsuzuki C., Rodrigues C.R., Mayer M.P. Genotypic and phenotypic analysis of Streptococcus mutans from different oral cavity sites of caries-free and caries-active children. Oral Microbiol. Immunol., 2007, vol. 22, no. 5, pp. 313-319. doi: 10.1111/j.1399-302X.2007.00361.x
11. Nakano K., Ooshima T. Serotype classification of Streptococcus mutans and its detection outside the oral cavity. Future Microbiol., 2009, vol. 4, no. 7,pp. 891-902. doi: 10.2217/fmb.09.64
12. Oral Health Survey. Basic methods; 5th ed. Geneva: World Health Organization, 2013. 70p.
13. Palmer E.A., Nielsen T., Peirano P., Nguyen A.T., Vo A., Nguyen A., Jackson S., Finlayson T., Sauerwein R., Marsh K., Edwards I., Wilmot B., Engle J., Peterson J., Maier T., Machida C.A. Children with severe early childhood caries: pilot study examining mutans streptococci genotypic strains after full-mouth caries restorative therapy. Pediatr. Dent., 2012, vol. 34, no. 2, pp. 1-10.
14. Palmer E.A., Vo A., Hiles S.B., Peirano P., Chaudhry S., Trevor A., Kasimi I., Pollard J., Kyles C., Leo M., Wilmot B., Engle J., Peterson J., Maier T., Machida C.A. Mutans streptococci genetic strains in children with severe early childhood caries: follow-up study at one-year post-dental rehabilitation therapy. J. Oral Microbiol., 2012, no. 4, pp. 167-173. doi: 10.3402/jom.v4i0.19530
15. Rincon-Rodriguez R.J., Parada-Sanchez M.T., Bedoya-Correa C.M., Arboleda-Toro D. Genetic diversity of Streptococcus mutans serotype c isolated from white spot and cavitated caries lesions from schoolchildren. Arch. Oral Biol, 2019, vol. 100, pp. 33-41. doi: 10.1016/j.archoralbio.2019.02.003
16. Struzycka I. The oral microbiome in dental caries. Pol. J. Microbiol., 2014, vol. 63, no. 2, pp. 127-135.
17. Suzuki S., Ukiya T., Kawauchi Y., Ishii H., Sugihara N. Decision tree analysis for factors associated with dental caries in school-aged children in Japan. Community Dent. Health., 2018, vol. 35, no. 4, pp. 247-251. doi: 10.1922/CDH_4409Suz.uki05
18. Zhou Q., Qin X., Qin M., Ge L. Genotypic diversity of Streptococcus mutans and Streptococcus sobrinus in 3-4-year-old children with severe caries or without caries. Int. J. Paediatr. Dent., 2011, vol. 21, no. 6, pp. 422-431. doi: 10.1111/j.1365-263X.2011.01145.x
