Rapid identification of beta-hemolytic streptococci by fluorescence in situ hybridization (FISH)

Poppert, Sven; Nickel, Daniel; Berger, Anja; Yildiz, Tatjana; Kaestner, Nicole; Mauerer, Stefanie; Spellerberg, Barbara

doi:10.1016/j.ijmm.2009.02.001

Cited by 8 publications

(7 citation statements)

References 26 publications

(23 reference statements)

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“…pneumoniae), betahemolytic, or nonhemolytic when describing their appearance and being able to be grouped into different serogroups via typing of Lancefield antigens. Species identification of cultured streptococci is traditionally performed either by biochemical methods, including bile solubility and antibiotic resistance patterns (optochin/bacitracin sensitivity), or by agglutination testing (98). While streptococcal taxonomy has been in a state of flux over the previous decade, the accurate identification of these organisms in infectious processes is essential in most cases.…”

Section: Streptococcimentioning

confidence: 99%

“…These organisms are important human pathogens, and their timely and accurate identification is paramount in the context of infection and is crucial to clinical diagnosis. As such, a number of phenotypic and molecular methods were developed for their identification (98). BHS include Lancefield antigenic group A (S. pyogenes), antigenic group B (S. agalactiae), antigenic group C (S. dysgalac-tiae and others), and group G (S. canis and others).…”

Section: Streptococcimentioning

confidence: 99%

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Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: a Fundamental Shift in the Routine Practice of Clinical Microbiology

et al. 2013

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SUMMARY Within the past decade, clinical microbiology laboratories experienced revolutionary changes in the way in which microorganisms are identified, moving away from slow, traditional microbial identification algorithms toward rapid molecular methods and mass spectrometry (MS). Historically, MS was clinically utilized as a high-complexity method adapted for protein-centered analysis of samples in chemistry and hematology laboratories. Today, matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) MS is adapted for use in microbiology laboratories, where it serves as a paradigm-shifting, rapid, and robust method for accurate microbial identification. Multiple instrument platforms, marketed by well-established manufacturers, are beginning to displace automated phenotypic identification instruments and in some cases genetic sequence-based identification practices. This review summarizes the current position of MALDI-TOF MS in clinical research and in diagnostic clinical microbiology laboratories and serves as a primer to examine the “nuts and bolts” of MALDI-TOF MS, highlighting research associated with sample preparation, spectral analysis, and accuracy. Currently available MALDI-TOF MS hardware and software platforms that support the use of MALDI-TOF with direct and precultured specimens and integration of the technology into the laboratory workflow are also discussed. Finally, this review closes with a prospective view of the future of MALDI-TOF MS in the clinical microbiology laboratory to accelerate diagnosis and microbial identification to improve patient care.

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Section: Streptococcimentioning

confidence: 99%

Section: Streptococcimentioning

confidence: 99%

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: a Fundamental Shift in the Routine Practice of Clinical Microbiology

et al. 2013

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“…As a result, screening of mothers before delivery is critical for GBS, not only for prevention but also to exclude unnecessary administration of antibiotics to the mother. There is currently FISH probes in development to rapidly identify GBS organisms; however, the appropriate culture media to be used is still being determined [83].…”

Section: Beta Hemolytic Streptococcimentioning

confidence: 99%

Fish

Forrest¹,

Mohammadi²,

Mohammadi³

2015

Modern Techniques for Pathogen Detection

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“…6,15,16 A culture-independent molecular method allowing the direct detection of pathogens in clinical samples is fluorescent in situ hybridization (FISH). 24,25,29 Fluorescence-labeled oligonucleotide probes bind specific regions of the target RNA/DNA. Furthermore, several probes labeled with different fluorophores can be combined into a single test and therefore different species can be detected simultaneously.…”

Section: Introductionmentioning

confidence: 99%

Identification of pathogens in mastitis milk samples with fluorescent in situ hybridization

et al. 2013

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Abstract. Traditionally, the bacteriological examination of mastitis milk samples is performed by culture followed by biochemical tests on the cultured bacteria to allow identification of the causative pathogen. Depending on the species involved, this classic identification is time-consuming compared to other techniques such as fluorescent in situ hybridization (FISH), a culture-independent method that utilizes oligonucleotides (labeled with a fluorophore) that are specific to a string of target DNA/RNA. In the current study, the applicability of FISH was evaluated for the detection of mastitis pathogens directly in milk samples. To remove interfering lipids and proteins from mastitis milk samples prior to FISH, a previously published enzymatic treatment with savinase was evaluated. FISH was performed using oligonucleotides specific for Staphylococcus aureus, Streptococcus agalactiae, Streptococcus uberis, Enterococcus faecalis, Enterococcus faecium, Escherichia coli, and Trueperella (Arcanobacterium) pyogenes. The enzymatic pretreatment and the sensitivity of FISH were evaluated using spiked whole milk samples and mastitis milk samples with bacterial loads of less than 10 3 up to 10 8 colony-forming units (CFU)/ml. Bacteria were reliably detected in milk samples with bacterial numbers of 10 6 CFU/ml or higher. However, bacteria present in numbers below 10 6 CFU/ml were not detectable in all cases. The ability of FISH to identify mastitis-causing pathogens directly in milk samples, and therefore earlier than classical culture methods, can supplement the classic diagnostic procedures for mastitis milk samples.

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Rapid identification of beta-hemolytic streptococci by fluorescence in situ hybridization (FISH)

Cited by 8 publications

References 26 publications

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: a Fundamental Shift in the Routine Practice of Clinical Microbiology

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: a Fundamental Shift in the Routine Practice of Clinical Microbiology

Fish

Identification of pathogens in mastitis milk samples with fluorescent in situ hybridization

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