微生物学技术

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 ...

Although accurate and rapid diagnosis of fusariosis is now required, morphological similarities among molds make it difficult to histologically differentiate Fusarium spp. from other molds. In this chapter, we present our in situ hybridization (ISH) technique as a valuable tool to id ...

Oleaginous yeast Yarrowia lipolytica is capable of accumulating large amount of lipids. There is a growing interest to engineer this organism to produce lipid-derived compounds for a variety of applications. In addition, biosynthesis of value-added products such as carotenoid and i ...

Mucor circinelloides is a β-carotene producing zygomycete amenable to metabolic engineering using molecular tools. The crtS gene of the heterobasidiomycetous yeast Xanthophyllomyces dendrorhous encodes the enzymatic activities β-carotene hydroxylase and ketolase, ...

Several fungal species, particularly some included in the Mucorales, have been used to develop fermentation processes for the production of β-carotene. Oxygenated derivatives of β-carotene are more valuable products, and the preference by the market of carotenoids from biological ...

The carotene producer fungus Mucor circinelloides is the zygomycete more amenable to genetic manipulations by using molecular tools. Since the initial development of an effective procedure of genetic transformation, more than two decades ago, the availability of new molecular ap ...

The Mucoral fungus Blakeslea trispora is used for the industrial production of β-carotene and lycopene. Two genetic techniques have been used to increase carotene accumulation: the isolation of mutants and the formation and segregation of heterokaryons. Because all life stages are mu ...

The zygomycete Blakeslea trispora produces high amounts of the general zygomycete β-carotene-derived sexual signal compounds, the trisporoids. These can be isolated from the culture medium and purified by extraction with organic solvents followed by thin layer chromatograph ...

Carotenoids are among the most widely distributed pigments in nature, and they are exclusively synthesized by plants and microorganisms. These compounds may serve a protective role against many chronic diseases such as cancers, age-related macular degeneration, and cardiovasc ...

Microorganisms are one of the greatest sources of metabolic and enzymatic diversity. In recent years, emerging recombinant DNA and genomic techniques have facilitated the development of new efficient expression systems, modification of biosynthetic pathways leading to new me ...

Yeasts of the genera Rhodotorula are able to synthesize different pigments of high economic value like β-carotene, torulene, and torularhodin, and therefore represent a biotechnologically interesting group of yeasts. However, the low production rate of pigment in these microorga ...

The orange pigmentation of the ascomycete fungi Neurospora and Fusarium is mainly due to the accumulation of neurosporaxanthin, a carboxylic apocarotenoid whose possible biotechnological applications have not been investigated. From the discovery of the first enzyme of the bio ...

DNA assembler enables design and rapid construction of biochemical pathways in a one-step fashion by exploitation of the in vivo homologous recombination mechanism in Saccharomyces cerevisiae. It has many applications in pathway engineering, metabolic engineering, combina ...

The yeasts Xanthophyllomyces dendrorhous (teleomorph) and Phaffia rhodozyma (anamorph) are of basidiomycetous affinity and have the unique property among yeasts of producing the carotenoid pigment astaxanthin. Astaxanthin imparts the attractive coloration to salmon ...