Methodology of microbiological analysis of wound swabs within the framework of modern concepts of wound infection process

Yarets, Yu. I.; Шевченко, Н. Ю.; Vf, Eremin

doi:10.17116/labs20211003133

Cited by 7 publications

(3 citation statements)

References 15 publications

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“…Также учитывали микроорганизмы, выделенные качественным методом из среды обогащения. Дополнительно оценивали комплекс генотипических и фенотипических признаков, определяющих патогенный и персистентный потенциал бактерий [8]. С учетом полученных положительных результатов посевов в хронических ранах устанавливали основные состояния, определяющие стадии инфекционного процесса: колонизацию (n=39), критическую колонизацию (n=37) и инфекцию (n=29).…”

Section: материалы и методыunclassified

Features of the Local Immune Response in the Infectious Process of Acute and Chronic Wounds: Results of Immunohistochemical Studies of Cellular Immunity Factors

Ярец¹,

Zhandarov²

2023

Лабораторная Диагностика. Восточная Европа

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Цель. Осуществить иммуногистохимические исследования факторов клеточного иммунитета в биоптатах острых и хронических ран с учетом клинико-микробиологических характеристик раневой поверхности. Материалы и методы. Выполнен анализ относительного содержания (%) CD3+ (Т-лимфоциты), CD4+ (Т-хелперы) и CD8+ (Т-цитотоксические лимфоциты), CD20+ (В-лимфоциты), CD138+ (плазмоциты), CD68+ (макрофаги), CD15+ (нейтрофилы), CD34+ (эндотелий сосудов), CD57+ (натуральные киллеры), d2-40 (лимфатический эндотелий), Ki 67+ (маркер пролиферации), каспаза 3+ (маркер апоптоза), S100 (маркер воспаления), COX-2 (маркер воспаления) клеток по результатам иммуногистохимического исследования биоптатов грануляционной ткани пациентов с острыми ранами (ОР, n=30, срок раны от 5 до 21 суток) и хроническими ранами (ХР, n=120, срок раны более 3 недель). Оценивали также клинический статус раны на предмет наличия признаков воспаления и выполняли микробиологический посев раневого отделяемого. Результаты. Более высокие значения иммунорегуляторного индекса (отношение CD4+/CD8+) и маркера пролиферации Ki 67+ регистрировались в острых ранах сроком от 5 до 21 суток, более низкие – в хронических ранах сроком более 22 суток и существенно не изменялись в зависимости от клинико-микробиологического состояния ран. Дополнительным признаком локального иммунного ответа являлось количество нейтрофилов, преобладающее в хронических ранах. Воспалительный статус ран и присутствие микроорганизмов, переход от стадии колонизации к стадии критической колонизации и инфекции характеризовались увеличением относительного содержания в ранах Т-лимфоцитов и их основных субпопуляций (CD4+ и CD8+ клеток), В-лимфоцитов, нейтрофилов и макрофагов, более высокими значениями S100 и COX-2. При выделении из хронических ран монокультур S. aureus в биоптатах в меньшем количестве обнаруживались В-лимфоциты, натуральные киллеры и макрофаги по сравнению с их содержанием в ранах, в которых этиологическими факторами инфекционного процесса были P. aeruginosa и A. baumannii. Выводы. Определение клеточных факторов локального иммунитета является перспективным направлением для дифференциальной диагностики острой и хронической раны, а особенности их изменения позволяют установить стадию инфекционного процесса. Purpose. To analyze the results of immunohistochemical studies of biopsy specimens from acute and chronic wounds taking into account their clinical and microbiological characteristics. Materials and methods. The relative content (%) of CD3+ (T-lymphocytes), CD4+ (T-helpers) and CD8+ (T-cytotoxic lymphocytes), CD20+ (B-lymphocytes), CD138+ (plasmocytes), CD68+(macrophages), CD15+(neutrophils), CD34+(vascularendothelium), CD57+ (natural killer cells), d2-40 (lymphatic endothelium), Ki 67+ (proliferation marker), caspase 3+ (apoptosis marker), S100 (inflammatory marker), COX-2 (inflammatory marker) cells was analyzed according to the results of immunohistochemical study of granulation tissue biopsy specimens from patients with acute wounds (n=30, wound duration from 5 to 21 days) and chronic wounds (n=120, wound duration more than 3 weeks). The clinical status of the wound was also assessed for signs of inflammation, and microbiological culture of wound swabs was performed. Results. Higher values of the immunoregulatory index (the ratio of CD4+/CD8+ lymphocytes) and the proliferation marker Ki 67 were recorded in acute wounds for a period of 5 to 21 days, lower values – in chronic wounds for a period of more than 22 days, and did not significantly change depending on the clinical and microbiological state of wounds. An additional feature was the number of neutrophils predominating in chronic wounds. The inflammatory status of wounds and the presence of microorganisms, as well as switch from colonisation stage to critical colonisation and infection stage were characterised by an increase in T-lymphocytes relative content and their major subpopulations (CD4+ and CD8+ cells), B-lymphocytes, neutrophils and macrophages, higher values of S100 and COX-2 in wounds. When S. aureus monocultures were isolated from chronic wounds, B-lymphocytes, natural killer cells and macrophages were lower in biopsy specimens compared to their content in wounds in which P. aeruginosa and A. baumannii were etiologic factors of the infectious process. Conclusions. Immunohistochemical cell markers are promising objective criteria for differential diagnosis of acute and chronic wounds, allowing to establish the infectious process stage.

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Section: материалы и методыunclassified

Features of the Local Immune Response in the Infectious Process of Acute and Chronic Wounds: Results of Immunohistochemical Studies of Cellular Immunity Factors

Ярец¹,

Zhandarov²

2023

Лабораторная Диагностика. Восточная Европа

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“…Для гистологического исследования выполняли биопсию раны, при этом в образец включали участок здоровой ткани и зону наиболее выраженных гнойно-некротических изменений грануляционной ткани. [19]. Исследование выполнялось однократно, на момент поступления пациентов.…”

Section: материалы и методыunclassified

“…Микробиота ран была представлена монокультурами и ассоциациями, частота встречаемости которых не различалась на различных сроках существования. Учитывая соблюдение необходимых правил преаналитического этапа получения раневого отделяемого, можно исключить возможность обнаружения контаминантов в результате микробиологического посева [19]. Выделенные изоляты будут иметь клиническое значение в плане нарушения процесса заживления, что, несмотря на отсутствие явного воспалительного статуса, подтверждается клиническими особенностями грануляционной ткани (таблица 2).…”

Section: результаты и обсуждениеunclassified

Colonized, critically colonized and infected wounds: differentiation using clinical and microbiological and morphological methods of investigation

Yarets

Славников

Дундаров

2022

jour

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Objective. To analyze the clinical and microbiological and morphological features of wounds of different duration, allowing to differentiate the stages of the infectious process and to determine recommendations for further tactics of preoperative preparation.Materials and methods. Clinical and microbiological and morphological assessment of the state of acute and chronic wounds in 313 patients was performed.Results. Colonized wounds were characterized by the presence of pathological signs (atrophy, cicatricial changes) in fine-grained granulations, the frequency of which increased from 38.5% for wounds lasting 22–28 days up to 85.1% for wounds lasting more than 2 months (χ2 = 14.0 ; p=0.003). Infected wounds were more often detected at terms of 22–28 days (24.6%, vs 3.9% for wounds more than 2 months old, χ2=40.51; p<0.001). Critically colonized wounds made up with coarse-grained granulations were detected at all stages of wound existence (from 21.6 to 32.8%). The analysis of biopsy specimens revealed the initial signs of proliferation disorders in 22-28 day wounds, which justifies the need to classify them as chronic. Second phase reparation failure was detected in all cases of chronic wounds; the severity of the signs of purulent inflammation increased depending on the stage of the infectious process.Conclusion. Differentiation of the stages of the infectious process in wounds using the clinical and microbiological and morphological research methods is necessary for the determination of the indications for the use of local wound debridement and systemic antibiotic therapy at the stage of wound preparation for plastic closure.

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Substantiation of tactics of chronic wound bed preparation at various stages of the infectious process on the basis of the analysis of the results of microbiological, clinical and morphological parameters

Yarets,

Slavnikov

2023

Vescì Nac. akad. navuk Belarusì, Ser. med. navuk

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The chronic wounds (CW) bed preparation requires an integrated approach using wound assessment, microbiological and histological analysis, which will avoid skin graft failure.The aim was to evaluate the possibility of using microbiological and morphological parameters of CW assessment to substantiate the choice of tactics of wound bed preparation to skin grafting (SG).Morphological and microbiological indicators of CW (n = 229) at various stages of the infectious process were analyzed. To prepare CW to SG, the following treatment methods were used: dressings, vacuum therapy (VT), mechanical debridement (MD), and ultrasound debridement (UD).The criteria that determine the possibility of CW preparation “under a bandage” in combination with MD are: the absence of inflammation signs and the normal state of granulations (red-pink color, firm and moist), a negative result of wound swabs, the histological result of Si 1/Si 2 and Sp 1/Sp 2. For CW performed with pathologically altered granulations colonized by potential pathogens and having Sp 3, Si 2/Si 3 criteria, the use of 2 UD procedures is indicated (1st in combination with MD, 2nd before SG), between which VT is applied. Critically colonized wounds showing the signs of Sp 3 in combination with Si 1 or Si 2, from which microorganisms are isolated in an amount of ≤105 CFU/ml and are characterized by persistent properties, are an indication for VT, which is performed between two UD procedures. Clinical infection signs, the presence of bright red friable and bulge granulations in the wound, morphological signs of Si 3/Si 2 and Sp 1/Sp 2 are indications for the use of 2 UD procedures in combination with systemic antibiotic therapy based on wound swab culture results.Microbiological and morphological parameters characterize the CW state at various stages of the infectious process (colonization, critical colonization, infection) and are recommended as criteria for choosing tactics for wound bed prepara tion to SG.

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Methodology of microbiological analysis of wound swabs within the framework of modern concepts of wound infection process

Cited by 7 publications

References 15 publications

Features of the Local Immune Response in the Infectious Process of Acute and Chronic Wounds: Results of Immunohistochemical Studies of Cellular Immunity Factors

Features of the Local Immune Response in the Infectious Process of Acute and Chronic Wounds: Results of Immunohistochemical Studies of Cellular Immunity Factors

Colonized, critically colonized and infected wounds: differentiation using clinical and microbiological and morphological methods of investigation

Substantiation of tactics of chronic wound bed preparation at various stages of the infectious process on the basis of the analysis of the results of microbiological, clinical and morphological parameters

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