microbiological rapid on-site evaluation using dq-staining for clinical decisions

Aprimorando o tratamento da infecção pulmonar usando o M-ROSE

The technique of rapid on-site evaluation (ROSE) is a highly efficient method employed in the field of pulmonary disease procedures. It enables real-time sampling and diagnostic intervention, facilitating immediate cytological analysis1. This innovative approach involves imprinting a portion of the tissue specimen onto a slide while preserving its integrity. One of the key advantages of ROSE lies in its capacity to facilitate prompt interpretation of clinical information through specialized microscopy techniques. This encompasses the analysis of cell morphology, classification, quantification, determination of constituent ratios, assessment of arrangement, correlation analysis, evaluation of background, and identification of foreign objects. By integrating all this data with the patient's clinical information, ROSE plays a pivotal role in evaluating sampling adequacy and guiding real-time interventional procedures and techniques2.
As a subsidiary of ROSE technology, M-ROSE technology primarily focuses on acquiring the microbiological background of target lesions rather than discerning between benign and malignant cells3,4. On one hand, M-ROSE enables the microscopic identification of pathogens such as Aspergillus, Cryptococcus, Pneumocystis, and Candida5. On the other hand, it holds significant guiding implications in assessing respiratory specimen quality, distinguishing infectious from non-infectious diseases, discriminating infection from contamination, as well as evaluating infection severity and prognosis6,7. For instance, within a respiratory specimen, the coexistence of bacteria exhibiting identical morphology alongside infiltrating inflammatory cells indicates an infection; conversely, the presence of multiple morphologically diverse bacteria accompanied by epithelial cells suggests contamination. The capacity to comprehensively analyze clinical information and predict outcomes renders M-ROSE an invaluable tool in pulmonary disease procedures.
In conclusion, ROSE serves as a crucial cytological carrier that significantly enhances the efficiency and precision of diagnosing lung diseases. Its multifaceted capabilities contribute to improved patient outcomes by ensuring timely intervention and facilitating accurate diagnosis through real-time sampling and diagnostic intervention techniques. However, it is important to note that these advancements rely on obtaining qualified samples. Herein, we present a standardized M-ROSE protocol encompassing slide preparation, staining techniques, and interpretation guidelines. This protocol serves as an invaluable reference for clinicians to establish accurate evaluation and treatment plans while facilitating decision-making regarding subsequent handling of target specimens.

Autor em destaque: Avançando na detecção de patógenos e avaliação de doenças em tempo real usando M-ROSE

Avaliação microbiológica rápida no local para doenças infecciosas pulmonares

Our intent is to introduce a real time rapid assessment method, M-ROSE to assist the physicians for formulating accurate treatment strategies for patient with pulmonary infections. Rapid cell staining technology and a high definition microscopic imaging system are currently used to advance research in the field. The number of clinical studies on M-ROSE is relatively limited.The biggest challenge is interpretation of M-ROSE results, which requires personnel, and trained personnel as it is difficult to visually identify pathogens and differentiate between infection and contamination. With a better understanding of pathogens causing infections in real time, physicians can avoid unnecessary broad spectrum antibiotics contributing to antibiotic resistance and as a treatment of pulmonary infections. M-ROSE is real time and highly efficient.M-ROSE enables the microscopic identification of pathogens such as aspergillus, cryptococcus, numerous and candida. It holds significant guiding implications in sizing respiratory basic quality, distinguishing infections from non-infectious disease, discriminating, infection from contamination, as well as evaluating infection, severity, and prognose.

Avaliação microbiológica rápida no local usando coloração DQ para decisões clínicas

Para começar, obtenha todos os materiais necessários para M-ROSE e coloque o conjunto completo de soluções de coloração Diff-Quik ou DQ em frascos de coloração de vidro com tampas seladas. Para a preparação da lâmina do rolo, extraia as partículas de tecido com uma agulha de seringa descartável de 2,5 a cinco mililitros. Em seguida, espalhe uma área circular de espessura moderada da seção interna para a externa de 1/3 da lâmina de citologia.Mergulhe a lâmina na solução Diff-A por 10 a 30 segundos. Para remover o excesso de solução de Diff-A, coloque a lâmina no banho de corante PBS e balance-a levemente para cima e para baixo. Em seguida, sacuda suavemente qualquer tampão restante.Mergulhe a lâmina na solução Diff-B por 20 a 40 segundos. Em seguida, coloque o slide no tanque de tingimento de água e balance-o levemente para cima e para baixo. Finalmente, usando papel absorvente, limpe o líquido residual da lâmina e prossiga para o exame microscópico.O M-ROSE da amostra de biópsia broncoscópica do paciente usando coloração DQ identificou pneumocystis, caracterizado por uma estrutura clara, contraste acentuado e ASCUS.

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Research

JoVE Journal

Medicine

Microbiological Rapid On-Site Evaluation Using DQ-Staining for Clinical Decisions

41 visualizações.  Chongqing Hospital of Traditional Chinese Medicine. Para começar, obtenha todos os materiais necessários para M-ROSE e coloque o conjunto completo de soluções de coloração Diff-Quik ou DQ em frascos de coloração de vidro com tampas seladas. Para a preparação da lâmina do rolo, extraia as partículas de tecido com uma agulha de seringa descartável de 2,5 a cinco mililitros. Em seguida, espalhe uma área circular de espessura moderada da seção interna para a externa de 1/3 da lâmina de citologia.Mergulhe a lâmina na soluç...

Vídeo: Avaliação microbiológica rápida no local usando coloração DQ para decisões clínicas

Microbiological Rapid On-Site Evaluation for Pulmonary Infectious Diseases

The prompt initiation of empirical anti-infective therapy is crucial in patients presenting with unexplained pulmonary infection. Although imaging acquisition is relatively straightforward in clinical practice, its lack of specificity often necessitates additional time-consuming tests such as sputum culture, bronchoalveolar-lavage fluid culture, or genetic sequencing to identify the underlying etiology of the disease accurately. Moreover, the limited efficacy of empirical anti-infective treatment may contribute to antibiotic misuse. Recent advancements in interpreting microbial background on rapid on-site evaluation (ROSE) slides have enabled clinicians to promptly obtain samples through bronchoscopy (e.g., alveolar lavage, mucosal brushing, tissue clamp), facilitating bedside staining and interpretation that provides essential microbial background information. Consequently, this establishes a foundation for developing targeted anti-infection treatment and individualized drug therapy plans. With a better understanding of which pathogens are causing infections in real-time, physicians can avoid unnecessary broad-spectrum antibiotics contributing to antibiotic resistance. Establishing a rapid and standardized M-ROSE workflow within respiratory medicine departments or intensive care units will greatly assist physicians in formulating accurate treatment strategies for patients, which holds significant clinical implications.

The protocol here demonstrates a fast and standardized microbiological rapid on-site evaluation (M-ROSE) workflow, including three steps: slide making, staining, and interpretation. This protocol will help physicians make rapid clinical decisions.