实验基础

Comparative genomic hybridization analyses have contributed greatly to our understanding of bacterial evolution, population genetics, and pathogenesis. Here, we describe a robust protocol for microarray-based comparison of genome content, which could be applied to any bact ...

Bacteria communicate with other members of their community through the secretion and perception of small chemical cues or signals. The recognition of a signal normally leads to the expression of a large suite of genes, which in some bacteria are involved in the regulation of virulence factors, ...

The lack of a system for genetic manipulation has hindered studies on the molecular pathogenesis of relapsing fever Borrelia. The focus of this chapter is to describe selectable markers, manipulation strategies, and methods to electro-transform and clone wild-type infectious Borr ...

Borrelia burgdorferi, the causative agent of Lyme disease, is an obligate parasite that cycles between vertebrate hosts and tick vectors. Attempts to understand the genetic factors that allow B. burgdorferi to sense, adapt to, and survive in different environments have been limited by a rel ...

Exopolysaccharides play a crucial role in the formation of biofilms and biofilm resistance to antimicrobials and innate host defense. Here we describe methods to analyze and quantify polysaccharide intercellular adhesin (PIA), a biofilm exopolysaccharide made of N-acetylglu ...

We developed PCR assays to detect and quantitate Yersinia pestis, the bacterial agent of plague, in flea vector and mammalian host tissues. Bacterial numbers in fleas, fleabite sites, and infected lymph nodes were determined using real-time PCR with primers and probes for a gene target on a multi ...

Intracellular bacterial pathogens have evolved sophisticated strategies to survive and proliferate within cells of their hosts. Studying their intracellular life cycle is key to understanding virulence and requires methodologies that can identify the compartments in whi ...

Anaplasma phagocytophilum is the etiologic agent of granulocytic anaplasmosis, a tick-borne, zoonotic, emerging infectious disease. A. phagocytophilum is an obligate intracellular pathogen that primarily resides within membrane-bound, cytoplasmic vacuoles of host ne ...

Electron microscopy of bacterial pathogens and interactions between bacteria and host cells and tissues provides valuable insights into structural and molecular properties and processes involved in pathogenesis. Applications for electron microscopy in bacterial path ...

Coxiella burnetii, the agent of Q fever, is an obligate intracellular bacterium that has a tropism for cells of the mononuclear phagocyte system. Following internalization, C. burnetii remains in a phagosome that ultimately matures into a vacuole with lysosomal characteristics that s ...

Whole body biophotonic imaging (BPI) is a technique that has contributed significantly to the way researchers study bacterial pathogens and develop pre-clinical treatments to combat their ensuing infections in vivo. Not only does this approach allow disease profiles and drug effic ...

Humana Press, Totowa, NJ This chapter describes methods for using non-human primates as a model of group A streptococcal (GAS) pharyngitis. This model has been used successfully to study host–pathogen interactions occurring during pharyngeal GAS infections. The protocol as describ ...

Staphylococcus aureus nasal colonization is a well-known risk factor for development of S. aureus infections in humans, but despite this established association, we are only beginning to understand the factors, both host and pathogen, that play a role in the colonization of the nares by S. aure ...

The continued increase in antibiotic resistance among bacterial pathogens, coupled with a decrease in infectious disease research among pharmaceutical companies, has escalated the need for novel and effective antibacterial chemotherapies. While current agents have emer ...

The analyses of numerous prokaryotic and eukaryotic genomes have revealed the presence of variable number tandem repeats (VNTRs). VNTR analysis is currently widely used to sub-speciate many bacterial, fungal, and viral pathogens and has facilitated a number of molecular epidemiol ...

The use of reverse genetics to generate recombinant viruses allows the researcher to investigate the exact functional significance of particular viral genes during the virus life cycle, by means of their deletion or modification in the viral genome. These studies can extend to the introdu ...

Human cytomegalovirus (HCMV), a double-stranded DNA virus in the herpesvirus family, is a ubiquitous virus that infects greater than 40–60% of the general population and up to 100% within some subpopulations and/or geographic areas (1). HCMV has a complex pathobiology because infection of i ...

Several diagnostic tools are available for the identification of acute and latent viral infections. Although newly developed nucleic acid amplification methods, such as the polymerase chain reaction (PCR), have proved to be very useful diagnostic procedures, conventional meth ...

The process of virus entry accomplishes the delivery of the viral genetic information into the cell so that replication can take place. Entry of enveloped viruses into mammalian cells requires that the virus attach to the host cell surface through an interaction between an envelope compone ...

As with numerous other branches of science, the study of human cytomegalovirus (HCMV) infection has been revolutionized by the polymerase chain reaction (PCR) method first devised by Mullis and Faloona (1). PCR allows the in vitro amplification of HCMV DNA sequences by the simultaneous pri ...