实验基础

In order to assess the diversity and function of microbial communities most effectively, molecular assays need to be designed that target the phylogenetic markers and functional genes that are key to major ecological processes and microorganisms. A streamlined design process is pres ...

Magnetoresistive bioassays, in which the traditional optical labels are replaced by magnetic labels, hold the promise of increased response speed, sensitivity, and portability in the detection system. These properties make these systems ideal for the monitoring of microbiolog ...

We report on the development of a sandwich hybridization assay on an electrode surface-based sensor device for the detection of toxic algae. This DNA (rRNA)-based sensor uses an electrochemical detection of a target sequence by means of its hybridization to a capture probe, with an attachment ...

Marine toxins need a continuous monitoring system that is usually implemented in most producer and consumer countries. This is needed to protect consumers, as marine toxins are very toxic compounds. Up to now, the mouse bioassay has been the method of choice, but there is a legal requirement in Euro ...

An IISA-ATP (integrated in situ analyzer for ATP) was developed for the quantitative determination of ATP (adenosine-5′-triphosphate) in ocean environments, including the deep-sea. Total (intracellular and dissolved) ATP was quantitatively determined using a simple lucifer ...

Invasive fungal infections (IFI) are a common cause of life-threatening events in immunocompromised patients. Early detection and identification of the fungal pathogen is an important prerequisite for timely onset of the most appropriate treatment. Methods based on fungal cultu ...

The multiplexing qualities of padlock probes and Luminex™ technology combined with the well-established quantitative feature of qPCR were the base for a ten-plex fungal detection protocol that quantitatively reveals ten different fungal species in a single experiment. Padlock p ...

The polymerase chain reaction is a powerful molecular tool for the detection and analysis of very small amounts of DNA. Today, hybridization probes are often used in real-time PCR for more sensitive and specific detection of pathogens and for determination of gene regulation or mutation ana ...

Rapid and quantitative detection of Aspergillus from clinical samples may facilitate an early diagnosis of invasive pulmonary aspergillosis (IPA). As nucleic acid-based detection is a viable option, we demonstrate that Aspergillus burdens can be rapidly and accurately detected ...

Fluorescence in situ hybridization (FISH) has been widely used for the detection and identification of microorganisms in their natural environments. In this chapter we describe the use of a simple FISH-based protocol to detect and identify clinically relevant yeast species in culture ...

The identification of fungal pathogens, though continuously improving, is still time-consuming and often inadequate for ensuring an early targeted therapy, which may be crucial for the treatment of invasive mycoses. Here, we describe a DNA-microarray system based on the arrayed-pri ...

The rapid and inexpensive extraction of fungal genomic DNA that is of sufficient quality for molecular approaches is central to the molecular identification, epidemiological analysis, taxonomy, and strain typing of pathogenic fungi. Although many commercially available and in ...

Fungi and yeasts are critical causes of acute infection. As such, the detection and identification of these organisms are crucial in the diagnosis of affected patient populations. There is a vast array of commercial tests currently available for diagnostic purposes. These vary from tradi ...

The incidence of invasive fungal infections (IFIs) has seen a marked increase in the last two decades. This is especially evident among transplant recipients, patients suffering from AIDS, in addition to those in receipt of immunosuppressive therapy. Worryingly, this increased incid ...

Pneumocystis jirovecii is an opportunistic pathogen responsible for severe pneumonia in immunocompromised patients. Flow cytometry (FC) is a method widely used in different areas of clinical diagnosis like hematology and immunology. Recently it has started to be used in microbio ...

Multiplex, real-time PCR has become an invaluable tool for the rapid identification of pathogens in clinical specimens enabling earlier and more targeted management of antimicrobial therapy. In this chapter, we describe the methodology behind a novel multiplex-tandem PCR (MT-PCR) ...

Molecular biological detection and quantification of fungal DNA targets today relies mainly on the �application of the polymerase chain reaction (PCR). However, this well-recognized technique necessitates the use of highly purified DNA, in a well-equipped lab environment by trai ...

Hyperbranching Rolling Circle Amplification (HRCA) is a technique derived from Rolling Circle Amplification (RCA) in which DNA polymerase replicates circularized oligonucleotide probes under isothermal conditions with either linear or geometric kinetics. Since its fi ...

Surface plasmon resonance (SPR)-based DNA biosensors have been shown to be rapid, label-free, and selective tools for the detection of PCR products. Here, we describe an SPR sensor based on DNA hybridization for the detection of Fusarium culmorum, a fungal pathogen of wheat. A 0.57 kb DNA fragment of F. ...

Chip-based flow cytometry is a rather new method that offers an easy, fast opportunity for examination of yeasts, such as Candida cells. In our study cell-chip technology was tested with ATCC Candida strains to determine their viability and susceptibility against antifungal agents, amph ...