A role of methicillinresistant Staphylococcus aureus strains and related molecular genetic features in developing purulent-necrotic forms of the diabetic foot syndrome

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Abstract

Purulent-necrotic  complications in patients with diabetic foot syndrome is one of the main causes resulting in amputation and disability, or even lethal outcome. Our study was aimed at investigating a role played by MRSA and related molecular genetic features, as well as antibiotic resistance in developing purulent-necrotic forms of the diabetic foot syndrome in Krasnoyarsk, in the 2010–2016 period. A microbiota profile related to purulent-necrotic complications, antibiotic susceptibility, as well as the molecular genetic features of methicillin-resistant Staphylococcus aureus were examined in 240 patients with diabetic foot syndrome. A bacteriological method was used to investigate microbiota profile related to purulent complications. Antibiotic sensitivity was analyzed by disc-diffusion method; staphylococcal antibiotic sensitivity was evaluated by screening, PCR, solid medium serial dilution, in accordance with the CLSI and EUCAST international recommendations. Genotyping and examining molecular genetic features were performed by using PCR, M-PCR, and sequencing. The data were analyzed by using WHONET (WHO) software. Significance level was set at p <0.05. It was found that microbiota profile linked to purulent-necrotic forms of the diabetic foot syndrome was presented by various Gram-negative microorganisms including Enterobacteriaceae spp. and non-fermenting bacteria accounting for 34.4% and 19.1%, respectively, as well as Gram-positive microorganisms found in 46.5% cases, including S. aureus (18.4% cases). Moreover, microorganisms were characterized by multiresistance to diverse antimicrobial drugs: percent-age of BLDS- and MBL-producers as well as MRSA comprised 36.4%, 30.3%, and 36.4%, respectively. Further, MRSA ST239/spa3(t037)/agr1/SCCmecIII.1.1.2(IIIA)/coaIV/tst+  clone dominated in patients with purulent-necrotic  forms of diabetic foot syndrome admitted to the right-bank and left-bank hospitals in the city of Krasnoyarsk that was characterized by a high virulence level and multidrug resistance. Next, subdominant MRSA genetic variants were presented by ST8/spa1(t008)/agr1/SCCmecIV.3.1.1/CoaIII and ST12/spanew(t156)/agr1/SCCmecUT/coaIorVII characterized by resistance to 1–2 groups of antimicrobials apart from β-lactams. Importantly, similar MRSA genetic variants earlier isolated in patients with other nosological entities common in this geographic region were also mainly verified in patients with purulent-necrotic forms of diabetic foot syndrome admitted to Krasnoyarsk hospitals. In particular, it was demonstrated that MRSA isolates predominantly belonging to the same genetic variants were detected in patients with purulent-necrotic forms of diabetic foot syndrome hospitalized both at the right-bank and left-bank hospitals of the city of Krasnoyarsk. Thus, long-term hospitalization of patients with purulent-necrotic forms of diabetic foot syndrome as well as subsequent repeated admittance to other hospitals in the city of Krasnoyarsk facilitates inter-hospital bacterial strain transmission.

About the authors

O. E. Khokhlova

Krasnoyarsk State Medical University named after professor V.F. Voyno-Yasenetsky

Author for correspondence.
Email: khokhlovaol@mail.ru
ORCID iD: 0000-0002-2829-5117

Khokhlova OlgaEvgenievna - PhD (Biology), Associate Professor, Department of Microbiology named after B.I. Zelmanovich, Krasnoyarsk State Medical University named after professor V.F. Voyno-Yasenetsky.

660022, Krasnoyarsk, Partizan Zheleznyak str., 1.

Phone: +7 (391) 220-13-61; +7 (908) 018-99-84 (mob.).

Russian Federation

Ya. Iwao

International Medical Education and Research Center

Email: indiansign_64@hotmail.com

Iwao Yasuhisa - PhD, Researcher, Laboratory of Molecular Bacteriology, International Medical Education and Research Center.

Niigata.

Japan

V. V. Kamshilova

Krasnoyarsk State Emergency Hospital named after N.S. Karpovich

Email: kamshylova@mail.ru

Kamshilova VeraVladimirovna - PhD (Biology), Bacteriologist, Krasnoyarsk State Emergency Hospital named after N.S. Karpovich.

Russian Federation

O. V. Teplyakova

Krasnoyarsk State Medical University named after professor V.F. Voyno-Yasenetsky; Krasnoyarsk State Municipal Clinical Hospital No. 7

Email: teplyakova-olga@yandex.ru
ORCID iD: 0000-0003-0005-3316

Teplyakova OlgaValerievna - PhD (Medicine), Associate Professor, Department of General Surgery named after M.I. Gulman, Krasnoyarsk State Medical University named after professor V.F. Voyno-Yasenetsky; Surgeon, Krasnoyarsk State Municipal Clinical Hospital No. 7.

660022, Krasnoyarsk, Partizan Zheleznyak str., 1.

Russian Federation

A. I. Motova

Krasnoyarsk State Emergency Hospital named after N.S. Karpovich

Email: tihotay@mail.ru

Motova AnastasiaIvanovna - Clinical Pharmacologist, Krasnoyarsk State Emergency Hospital named after N.S. Karpovich.

Russian Federation

A. I. Drobushevskaya

Krasnoyarsk State Municipal Clinical Hospital No. 7

Email: annushkadoc@mail.ru

Drobushevskaya AnnaIvanovna - PhD (Medicine), Surgeon, Krasnoyarsk State Municipal Clinical Hospital No. 7.

Russian Federation

O. V. Perianova

Krasnoyarsk State Medical University named after professor V.F. Voyno-Yasenetsky; Krasnoyarsk State Municipal Clinical Hospital No. 7

Email: perianova@mail.ru

Perianova OlgaVladimirovna - PhD (Biology), Head of Department of Microbiology named after B.I. Zelmanovich, Krasnoyarsk State Medical University named after professor V.F. Voyno-Yasenetsky; Director of Russian-Japan Center of Microbiology, Мetagenomics and Infectious Diseases, Krasnoyarsk State Medical University.

660022, Krasnoyarsk, Partizan Zheleznyak str., 1.

Russian Federation

Yu. S. Vinnik

Krasnoyarsk State Medical University named after professor V.F. Voyno-Yasenetsky

Email: yuvinnik@yandex.ru

Vinnik Yury Semenovich - PhD, MD (Medicine), Professor, Head of Department of General Surgery named after M.I. Gulman, Krasnoyarsk State Medical University named after professor V.F. Voyno-Yasenetsky.

660022, Krasnoyarsk, Partizan Zheleznyak str., 1.

Russian Federation

N. K. Potkina

Krasnoyarsk State Medical University named after professor V.F. Voyno-Yasenetsky; Russia-Japan Center of Microbiology, Мetagenomics and Infectious Diseases, Krasnoyarsk State Medical University

Email: potkinanad@mail.ru

Potkina Nadezhda Konstantinovna - Researcher, Krasnoyarsk State Medical University named after professor V.F. Voyno-Yasenetsky;  Researcher, Russian-Japan Center of Microbiology, Мetagenomics and Infectious Diseases, Krasnoyarsk State Medical University.

660022, Krasnoyarsk, Partizan Zheleznyak str., 1.

Russian Federation

D. E. Zdzitowiecki

Krasnoyarsk State Medical University named after professor V.F. Voyno-Yasenetsky; Krasnoyarsk State Emergency Hospital named after N.S. Karpovich

Email: zdz64@mail.ru
ORCID iD: 0000-0002-2498-3844

Zdzitowiecki Dmitry Eduardovich - PhD, MD (Medicine), Associate Professor, Head of the Department of Surgical Pathology named after Yu.M. Lubenskiy, Krasnoyarsk State Medical University named after professor V.F. Voyno-Yasenetsky; Surgeon, Krasnoyarsk State Municipal Clinical Hospital No. 7.

660022, Krasnoyarsk, Partizan Zheleznyak str., 1.

Russian Federation

T. Yamamoto

Russia-Japan Center of Microbiology, Мetagenomics and Infectious Diseases, Krasnoyarsk State Medical University; International Medical Education and Research Center

Email: tatsuoy@imerc.jp

Yamamoto Tatsuo - PhD, Professor, Curator of Russian-Japan Center of Microbiology, Мetagenomics and Infectious Diseases, Krasnoyarsk State Medical University; Director of International Medical Education and Research Center.

Niigata.

Japan

References

  1. Бахарев И.В., Редькин Ю.А. Синдром диабетической стопы: диагностика, лечение, профилактика. Качество жизни // Медицина. 2003. № 1. С. 35–38.
  2. Гостев В.В., Калиногорская О.С., Попенко Л.Н., Черненькая Т.В., Науменко З.С., Ворошилова Т.М., Захарова Ю.А., Хох лова О.Е., Круглов А.Н., Ершова М.Г., Молчанова И.В., Сидоренко С.В. Антибиотикорезистентность метициллинрезистентных Staphylococcus aureus, циркулирующих в Российской Федерации // Антибиотики и химиотерапия. 2015. Т. 60, № 1–2. C. 3–10.
  3. Доброквашин С.В., Якупов Р.Р., Валеев А.З. Лечение больных с гнойно-некротическими осложнениями синдрома диабетической стопы // Акушерство. Гинекология. Эндокринология. 2011. Т. 54, № 6. С. 97–99.
  4. Митиш В.А., Пасхалова Ю.С., Ерошкин И.А., Галстян Г.Р., Блатун Л.А. Гнойно-некротические поражения при нейроишемической форме синдрома диабетической стопы // Хирургия. 2014. № 1. С. 48–53.
  5. Попова Т.Е., Шнайдер Н.А., Петрова М.М., Говорина Ю.Б., Николаева Т.Я. Диагностика нарушений микроциркуляции у пациентов с преимущественно сенсорным вариантом хронической воспалительной демиелинизирующей полиневропатии: пилотное исследование // Сибирское медицинское обозрение. 2015. № 1. С. 32–37.
  6. Привольнев В.В., Решедько Г.К., Савкин В.А., Кречикова О.И. Структура возбудителей и их антибиотикорезистентность при инфекциях нижних конечностей у больных сахарным диабетом // Клиническая микробиология и антимикробная химитерапия. 2009. Т. 11, № 1. С. 86–89.
  7. Рундо А.И. Современные аспекты этиологии и патогенеза синдрома диабетической стопы // Новости хирургии. 2015. Т. 23, № 1. С. 97–104.
  8. Токмакова А.Ю. Современная концепция диагностики и лечения хронических ран у больных с синдромом диабетической стопы // Сахарный диабет. 2009. № 1. С. 14-17.
  9. Фенотипическое определение Enterobacter – продуцентов β-лактамаз расширенного спектра: обзор и руководство по проведению испытаний. Европейское общество к линической микробиологии и инфекционных болезней // К линическая микробиология и инфекционные заболевания. 2008. Т. 14 (Прил. 1). С. 90–103.
  10. Bal A.M., Coombs G.W., Holden M.T., Lindsay J.A., Nimmo G.R., Tattevin P., Skov R.L. Genomic insights into the emergence and spread of international clones of healthcare-, community- and livestock-associated meticillin-resistant Staphylococcus aureus: Blurring of the traditional definitions. J. Glob. Antimicrob. Resist., 2016, vol. 95–101, pp. 8–34. doi: 10.1016/j.jgar.2016.04.004
  11. Сavanagh P., Lipsky B.A., Bradbury A.W., Botek G. Treatment for diabetic foot ulcers. Lancet, 2005, vol. 366, pp. 1725–1735. doi: 10.1016/S0140-6736(05)67699-4
  12. Drieux L., Brossier F., Sougakoff W., Jarlier V. Phenotypic detection of extended-spectrum β-lactamase production in Enterobacteriaceae: review and bench guide. Clin. Microbiol. Infect., 2008, vol. 14 (suppl. 1), pp. 90 –103. doi: 10.1111/j.1469-0691.2007.01846.x
  13. Echaniz-Aviles G., Velazquez-Meza M.E., Vazquez-Larios Mdel R., Soto-Noguerón A., Hernández-Dueñas A.M. Diabetic foot infection caused by community-associated methicillin-resistant Staphylococcus aureus (USA300). J. Diabetes, 2015, vol. 7 (6), pp. 891–892. doi: 10.1111/1753-0407.12324
  14. Gostev V., Kruglov A., Kalinogorskaya O., Dmitrenko O., Khokhlova O., Yamamoto T., Lobzin Y., Ryabchenko I., Sidorenko S. Molecular epidemiology and antibiotic resistance of methicillin-resistant Staphylococcus aureus circulating in the Russian Federation. Infect. Genet. Evol., 2017, vol. 53, pp. 189–194. doi: 10.1016/j.meegid.2017.06.006
  15. Hiramatsu K., Ito T., Tsubakishita S., Sasaki T., Takeuchi F., Morimoto Y., Katayama Y., Matsuo M., Kuwahara-Arai K., Hishinuma T., Baba T. Genomic basis for methicillin resistance in Staphylococcus aureus. Infect. Chemother., 2013, vol. 45, pp. 117–136. doi: 10.3947/ic.2013.45.2.117
  16. International working group on the classification of staphylococcal cassette chromosome elements (IWG-SCC). Classification of staphylococcal cassette chromosome mec (SCCmec): guidelines for reporting novel SCCmec elements. Agents Chemother., 2009, vol. 53, pp. 4961–4967. doi: 10.1128/AAC.00579-09
  17. Jeffcoate W.J., Harding K.G. Diabetic foot ulcers. Lancet, 2003, vol. 361, no. 9368, pp. 1545–1551. doi: 10.1016/S0140-6736(03)13169-8
  18. Kado C.I., Liu S.T. Rapid procedure for detection and isolation of large and small plasmids. J. Bacteriol., 1981, vol. 145, pp. 1365–1373.
  19. Khokhlova O.E., Hung W.C, Wan T.W. Iwao Y., Takano T., Higuchi W., Yachenko S.V., Teplyakova O.V., Kamshilova V.V., Kotlovsky Y.V., Nishiyama A., Reva I.V., Sidorenko S.V., Peryanova O.V., Reva G.V., Teng L.J., Salmina A.B., Yamamoto T. Healthcare- and community-associated methicillin-resistant Staphylococcus aureus (MRSA) and fatal pneumonia with pediatric deaths in Krasnoyarsk, Siberian Russia: unique MRSA’s multiple virulence factors, genome, and stepwise evolution. PLoS One, 2015, vol. 1, pp. 1–30. doi: 10.1371/journal.pone.0128017
  20. Kondo Y., Ito T., Ma X.X., Watanabe S., Kreiswirth B.N., Etienne J., Hiramatsu K. Combination of multiplex PCRs for staphylococcal cassette chromosome mec type assignment: rapid identification system for mec, ccr, and major differences in junkyard regions. Antimicrob. Agents Chemother., 2007, vol. 51, pp. 264–274. doi: 10.1128/AAC.00165-06
  21. Lindsay J.A. Genomic variation and evolution of Staphylococcus aureus. Int. J. Med. Microbiol., 2010, vol. 300, pp. 98–103. doi: 10.1016/j.ijmm.2009.08.013
  22. Morbach S., Müller E., Reike H., Risse A., Rümenapf G., Spraul M. Diabetic foot syndrome. Exp. Clin. Endocrinol. Diabetes, 2014, vol. 122, no. 7, pp. 416–424. doi: 10.1055/s-0034-1366455
  23. Nelson A., Wright-Hughes A., Backhouse M.R., Lipsky B.A., Nixon J., Bhogal M.S., Reynolds C., Brown S. CODIFI (Concordance in Diabetic Foot Ulcer Infection): a crosssectional study of wound swab versus tissue sampling in infected diabetic foot ulcers in England. BMJ Open, 2018, vol. 8 (1): e019437, pp. 1–12. doi: 10.1136/bmjopen-2017-019437
  24. Post V., Wahl P., Uckay I., Ochsner P., Zimmerli W., Corvec S., Loiez C., Richards R.G., Moriarty T.F. Phenotypic and genotypic characterisation of Staphylococcus aureus causing musculoskeletal infections. Int. J. Med. Microbiol., 2014, vol. 304 (5–6), pp. 565–576. doi: 10.1016/j.ijmm.2014.03.003
  25. Prevost G., Jaulhac B., Piemont Y. DNA fingerprinting by pulsed-field gel electrophoresis is more effective than ribotyping in distinguishing among methicillin-resistant Staphylococcus aureus isolates. J. Clin. Microbiol, 1992, vol. 30, pp. 967–973.
  26. Reveles K.R., Duhon B.M., Moore R.J., Hand E.O., Howell C.K. Epidemiology of methicillin-resistant Staphylococcus aureus diabetic foot infections in a large academic hospital: implications for antimicrobial stewardship. PLoS One, 2016, vol. 24, no. 11 (8): e0161658. doi: 10.1371/journal.pone.0161658
  27. Sorensen A.I., Toft N., Boklund A., Espinosa-Gongora C., Græsbøll K., Larsen J., Halasa T. A mechanistic model for spread of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) within a pig herd. PLoS One, 2017, vol. 12, no. 11: e0188429, pp. 1–18. doi: 10.1371/journal.pone.0188429
  28. Takano T., Hung W.C., Shibuya M., Higuchi W., Iwao Y., Nishiyama A., Reva I., Khokhlova O.E., Yabe S., Ozaki K., Takano M., Yamamoto T. A new local variant (ST764) of the globally disseminated ST5 lineage of hospital-associated methicillin-resistant Staphylococcus aureus (MRSA) carrying the virulence determinants of community-associated MRSA. Antimicrob. Agents Chemother., 2013, vol. 57, pp. 1589–1595. doi: 10.1128/AAC.01147-12
  29. Tenover F.C., Arbeit R., Archer G., Biddle J., Byrne S., Goering R., Hancock G., Hébert G.A., Hill B., Hollis R. Comparison of traditional and molecular methods of typing isolates of Staphylococcus aureus. J. Clin. Microbiol., 1994, vol. 32, pp. 407–415.
  30. Van der Zwaluw K., de Haan A., Pluister G.N., Bootsma H.J., de Neeling A.J., Schouls L.M. The carbapenem inactivation method (CIM), a simple and low-cost alternative for the Carba NP test to assess phenotypic carbapenemase activity in gram-negative rods. PLoS One, 2015, vol. 10, no. 3: e0123690, pp. 1–13. doi: 10.1371/journal.pone.0123690

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Copyright (c) 2019 Khokhlova O.E., Iwao Y., Kamshilova V.V., Teplyakova O.V., Motova A.I., Drobushevskaya A.I., Perianova O.V., Vinnik Y.S., Potkina N.K., Zdzitowiecki D.E., Yamamoto T.

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