微生物学技术

The use of DNA probes for the detection and identification of para sites has become a routine practice in many laboratories and the tech nique is rapidly becoming established as a diagnostic tool for field and clinical studies. DNA probes have been used in the identification of a large number of paras ...

Whole parasites or nucleic acids from these organisms can be immo bilized on filter supports, such as nylon or nitrocellulose. This allows many samples to be tested for the presence of a specific sequence by hybridization to a complementary DNA probe. In addition, the copy number of this sequence can ...

Trypanosoma cruzi is a protozoan flagellate that is transmitted to mammals by bloodsucking triatomine bugs. Transmission is not by the bite of the insect but by contamination of skin abrasions or mucous membranes with bug feces containing infective (metacyclic) trypomastigote form ...

The preparation of nucleic acids from schistosomes can be divided into several stages, beginning with the collection and cleaning of the parasite material, its subsequent mechanical disruption to release cellular contents, the separation of nucleic acids from that lysate and, final ...

Extraction of nucleic acids is fundamental to molecular genetic studies of parasitic organisms. DNA is required for gene bank construction and analysis of the genome, and RNA is needed for cDNA synthesis, analysis of transcription, and translation studies. Early methods of isolating DNA ...

The genus Leishmania includes species that are the causative agents of visceral, cutaneous, and mucocutaneous leishmaniasis. Infections caused by these organisms are common in both the Old and New Worlds, where they represent a major public health problem. As a result, Leishmania species ...

The World Health Organization estimates that between 16 and 18 million people are infected with Trypanosoma cruzi, the protozoan parasite that causes American trypanosomiasis, or “Chagas′ disease” (1). An additional 70 million people are thought to be at risk of infection with the parasit ...

The protozoan Trypanosoma brucei is the most destructive parasite of domestic livestock in sub-Saharan Africa. The parasite lives extracellularly in its two hosts; alternately in the bloodstream of a mammal, and in the midgut and subsequently the salivary glands of the tsetse fly (Glossi ...

Any biochemical analysis is usually made easier by the availability of large numbers of cells to be analyzed, and one of the reasons for the position held by Trypanusoma brucei as the best characterized parasite is the relative ease with which it can be cultured in the laboratory. The ability to culture ...

The scope of this book does not allow a complete description of the many techniques available for purification and treatment of reagents for facilitating immunoassays in general. There is a large amount of literature covering techniques, and this can be consulted for specific problems. The ...

In this chapter, the use of control charts to both continuously evaluate testing in individual laboratories as well as provide data for external monitoring is examined in detail. The data is based on the publication by D. E. Rebeski' et al., “Charting methods to monitor the operational performance ...

This chapter deals with control charts to monitor the performance of Indirect ELISAs. An Indirect ELISA kit for the detection of antibodies against Brucella is used to demonstrate the methods. Many of the features explained in Chapter 9 are relevant to this chapter; some repetition is intended, ...

Standard protocols for definite species identification of nontuberculous mycobacteria by phenotypic characteristics include complex and time-consuming procedures. Other methods based on lipid analysis, such as high-performance liquid chromatography (HPLC), thin ...

Tuberculosts (TB) IS the most sign&ant smgle mfecttous cause of morbrdtty and mortalrty, producing eight million new cases and killing three million people annually. It causes approximately one quarter of all avoidable adult deaths from mfectton (1–3).

Mycobacteria, like other bacteria, respond to environmental signals with changes in gene expression regulated at the level of transcription. The significance of gene-expression patterns in pathogens, such asMycobacterium tuberculosis, is that specific signals encounter ...

Mycobacterium bovis Bacille Calmette Gu�rin (BCG) has several features that make it desirable as a vaccine vehicle for foreign antigens. BCG is a live vaccine and stimulates a strong cell-mediated immune response, is extremely safe, and can be given at birth (1). Using recently developed tools to ...

Over the past 10 years, major advances have been made in the field of molecular mycobacterioiogy. These include the generation of shuttle vectors for use in both Escherichia coli and mycobacteria, electroporation protocols to introduce these plasmids into mycobacteria and promoter c ...

Bioluminescence is the product of two distinct enzymes, firefly luciferase and bacterial luciferase. The application of the firefly enzyme in the study of mycobacteria has been described by other groups (1,2) (see Chapter 31). The bacterial-luciferase enzyme is a drmer of approx 80 kDa, cons ...

Gene replacement is a powerful technique that exploits the innate ability of an organism to recombme homologous regions of DNA. It allows for the specrfic replacement of targeted genomic sequences with copies of those sequences carrying defined mutations, and therefore can facilitate ...

Much of the present knowledge of the bacterial cell emanates from studies of spontaneous mutants and mutants obtained as a result of random or directed mutagenesis. Spontaneous mutants in a specific gene can generally be Isolated only if the mutation leads to a selectable phenotype, e.g., muta ...