Applications of Flow Cytometry to Clinical Microbiology

Pieretti, Barbara; Masucci, Annamaria; Moretti, M.

doi:10.5772/37325

Cited by 4 publications

(5 citation statements)

References 34 publications

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“…Akım sitometrisi yöntemi, bakteriyolojide hem bakterilerin hızlı tanımlaması hem de antibiyotik duyarlılıklarının saptanmasında kullanılmaya başlanmıştır 23 . İşaretli antikorların kullanması ile balgam örneklerinden direkt olarak mikobakteriler üç saatten kısa sürede tespit edilebilmektedir 5 .…”

Section: Discussionunclassified

“…Akım sitometrisi ile aynı örnekte birden fazla mikroorganizma tanımlanması yapılabilmektedir 24 . Akım sitometrisi, özellikle standart duyarlılık test yöntemleri ile sonuç alınmasının uzun sürdüğü mikobakteri ve mantar gibi mikroorganizmaların antibiyotik duyarlılık sonuçlarının kısa sürede alınmasında da kullanılmakta-dır 23 . Bu yöntem ayrıca, antimikrobiyal ajanların bakteriyostatik ve bakterisidal etkilerini göstermekte de yararlıdır ve minimal bakterisidal konsantrasyon değerlerini de tespit edebilmektedir 6 .…”

Section: Discussionunclassified

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Rapid Detection of Extended-Spectrum Beta-Lactamases by Flow Cytometry Method

Duyan¹,

Kılıç²,

Yılmaz

et al. 2015

Mikrobiyol Bul

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ÖZEnterobacteriaceae üyelerinde görülen genişlemiş spektrumlu beta-laktamaz (GSBL), özellikle klinikte yaygın olarak kullanılan sefalosporin grubu antibiyotiklere direnç gelişmesine neden olan bir enzimdir. GSBL üretiminin erken ve doğru olarak tespit edilmesi, antimikrobiyal tedavi ve enfeksiyon kontrolü için önemlidir; ancak bu amaç için kullanılan yöntemler zaman alıcıdır (24-48 saat). Bu çalışmanın amacı, GSBL üreten bakterilerin kısa sürede tespit edilmesine imkan veren, akım sitometrisi temelli bir test yönteminin belirlenmesidir. Çalışmaya, 2012-2013 yılları arasında izole edilmiş olan, GSBL üreten 38 (29 Escherichia coli, 9 Klebsiella pneumoniae) ve GSBL üretmeyen 10 (5 E.coli, 5 K.pneumoniae) Enterobacteriaceae suşu dahil edilmiştir. İzolatların tanımlanması ve antibiyotik duyarlılık testleri, Phoenix TM 100 otomatize sistemi (Becton Dickinson, ABD) ile yapılmıştır. GSBL pozitif izolatlar polimeraz zincir reaksiyonu yöntemiyle bla TEM , bla SHV , bla CTX-M1 , bla CTX-M2 ve bla CTX-M9 beta-laktamaz genlerinin varlığı açısından araştırılmıştır. Toplam 38 izolatın 36'sında en az bir GSBL geni tespit edilirken, iki E.coli izolatında gen tespit edilmemiştir. Akım sitometrisi yöntemi kullanılarak, sefalosporin [seftazidim (CAZ) veya sefotaksim (CTX)] ve klavulanik asit (CLA) kombinasyonuyla muamele edildiğinde ölen hücre yüzdeleri, sadece sefalosporin (CAZ veya CTX) ile muamele sonrası ölen hücre yüzdelerine oranlanmıştır. CAZ ve CTX için CLA indeks değerleri (CAZ-CLA ve CTX-CLA indeksi) elde edilmiştir. İndeks değerin en az bir sefalosporin için 1.5'ten daha büyük olması GSBL pozitif olarak değerlendirilmiştir. Çalışmamızda, GSBL pozitif olan izolatların genotipik özelliklerine göre CTX-CLA indeksi ortalamaları 1.14 ile 7.22; CAZ-CLA indeks ortalamaları ise 0.85 ile 5.6 arasında bulunmuştur. GSBL pozitif 38 ve GSBL negatif 10 izolattan oluşan iki grup değerlen-dirildiğinde; hem CAZ-CLA hem de CTX-CLA indeksi için iki grup arasında istatistiksel olarak anlamlı fark olduğu saptanmıştır (p< 0.005). CTX-CLA ve CAZ-CLA indeksleri istatistiksel olarak karşılaştırıldığında ise, CTX-CLA indeksinin GSBL pozitifl eri saptayabilme özelliğinin daha iyi olduğu belirlenmiştir (p= 0.001).

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Section: Discussionunclassified

Rapid Detection of Extended-Spectrum Beta-Lactamases by Flow Cytometry Method

Duyan¹,

Kılıç²,

Yılmaz

et al. 2015

Mikrobiyol Bul

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“…Tabel 1. Some of the fluorescent molecules used to study microorganisms by flow cytometry (modified from., [15,16].…”

Section: Fcm Applications In Microbiologymentioning

confidence: 99%

Updates on the applications of flow cytometry in microbiology

2019

Lett Appl NanoBioSci

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Flow cytometry (FCM) was first developed for medical and clinical applications such as hematology and oncology. Although these areas still account for the vast majority of publications on this technique, during the past few years it has been also introduced in other areas, such as optimization and monitoring of biotechnological and environmental processes, pharmacology, toxicology, bacteriology and virology. In the food and drinks industries, the time required for conventional microbiology tests can lead to substantial delays in product release to the market. FCM has been used in conjunction with viability markers for rapid counting of yeast, molds and bacterial cells, including foodborne pathogens and microbial contaminants, in food products as well as for monitoring and improving the final products quality. FCM is an excellent tool, still unexplored in clinical microbiology, allowing for detection of cellular and non-cellular components in different clinical specimens, the study of antimicrobial activity, allowing for rapid and direct antimicrobial susceptibility testing and for the investigation of resistance mechanisms. Recent FCM developments important for addressing questions in environmental microbiology include the study of microbial physiology under environmentally relevant conditions, the development of new methods to identify active microbial populations and to isolate previously uncultured microorganisms and of high-throughput autofluorescence bioreporter assays. Moreover, the technological advancements will make possible the miniaturisation and automation of FCM devices, allowing to revolutionize their applications in the near future. The purpose of this minireview is to update the current applications of FCM in different fields of applied microbiology, and to highlight the main advantages and pitfalls for each of them.

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“…Flow cytometry has been proposed as rapid diagnostic tool in the fight against infection caused by bacteria, viruses, fungi and parasites. It can be used to isolate microorganisms, identify them, and determine antibodies to a particular parasite or pathogen, in different stages of the disease and in direct detection of essential components such as nucleic acids and proteins in clinical specimens (Pieretti et al 2012). It could also count number of parasites and evaluate the malaria-infected red cells (Jang et al, 2014).…”

Section: Flow Cytometric Technique (Fcm)mentioning

confidence: 99%

“…Flow cytometric assays use different staining methods to detect parasite infected cells, most of which use nucleic staining procedure because nucleic acids can be found only in malaria infected RBCs but not in normal RBCs .There are numerous stains used in FCM, such as; Hoechst 33258 (Brown et al, 1980), Acridine orange (Whaun et al, 1987), Thiazole orange, (Makler et al, 1983), Hydroethidine (Wyatt et al, 1991), YOYO-1 (Barkan, et al, 2000), SYBR Green I and CD235A (Pieretti et al, 2012). These DNA-targeting dyes have different target specificity and fluorescence intensity which allows for better separation of cellular populations (Rye et al, 1992) and they can be used simultaneously as in Jang et al (2014).…”

Section: Flow Cytometric Technique (Fcm)mentioning

confidence: 99%

Molecular Diagnostic Techniques for Malaria Infection - A Review

Yangora, Y.M.,

Usman, A.D.

2017

UJMR

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Malaria is the most threatening disease protozoal and a major health problem worldwide especially in developing countries. WHO recommended that for every case of suspected malaria, diagnostic test must be done to confirm the disease. A more advanced Diagnostic Techniques were developed to overcome the problem of conventional microscopy. These techniques are known as Molecular diagnostic techniques, and they detect specific sequence in DNA, RNA and proteins to provide clinical information for human pathogens including malaria parasites. There are several techniques involve in molecular diagnostics, some are however discussed in this review. They include Polymerase Chain Reaction (PCR), Loop-mediated isothermal amplification (LAMP), Flow Cytometric technique (FCM), Nucleic acid based sequence amplification (NASBA), and Luminex xMax technology. Among these techniques; LAMP technique is the best techniques that can be deployed in the field settings (clinical and rural settings) because of its simplicity, reliability, stability, detection method as well as point-of- care and confirmatory ability. On the other hand, PCR-based technique is more suitable for research purposes because it can be used to identify drug resistance, follow-up therapeutic response, and detect asymptomatic malaria carriers who may be targeted for treatment. Hence, molecular diagnostic techniques are most innovative science and technical implementations that can be used to diagnose malaria infection and to overcome the limitations.

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Applications of Flow Cytometry to Clinical Microbiology

Cited by 4 publications

References 34 publications

Rapid Detection of Extended-Spectrum Beta-Lactamases by Flow Cytometry Method

Rapid Detection of Extended-Spectrum Beta-Lactamases by Flow Cytometry Method

Updates on the applications of flow cytometry in microbiology

Molecular Diagnostic Techniques for Malaria Infection - A Review

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