实验基础

Loop-mediated isothermal amplification (LAMP) is an established nucleic acid amplification method offering rapid, accurate, and cost-effective diagnosis of infectious diseases. The LAMP assay requires 12–18 min for amplification with a single colony on selective agar from ch ...

Bacterial protein toxins are involved in a number of infectious and foodborne diseases and are considered as potential biological warfare agents as well. Their sensitive multiplex detection in complex environmental, food, and biological samples are an important although challe ...

Saliva is an ideal translational research tool and diagnostic medium and is being used in novel ways to provide molecular biomarkers for a variety of oral and systemic diseases and conditions. The ability to analyze saliva to monitor health and disease is a highly desirable goal for oral health pr ...

Salivary biomarkers for diagnostic and prognostic assessments have become increasingly well established in recent years. Salivary mRNA transcriptomic analyses create a new paradigm in the emerging field for noninvasive molecular diagnosis. In this chapter, we will overview t ...

Saliva harbors a wide spectrum of proteins that may reflect the health/disease status in the human body. Profiling of the proteins in saliva from a disease population can potentially yield valuable clinical parameters to be used for diagnosis and prognosis of the disease. Advances in proteo ...

Colonization of surfaces in the human body by microorganisms is an early, essential, step in the initiation of infectious disease. We have developed in vitro assays to investigate interactions between yeast or bacterial cells and human tissues, fluids, or prostheses. Such assays can be used to ...

The techniques for the establishment of primary gingival and periodontal ligament fibroblast cultures have been well established for over 30 years. It is only more recently, with the commercial availability of real-time PCR (RT-PCR) gene arrays that the expression profiles of up to 84 genes ...

Gene expression profiling, i.e., the systematic cataloging of messenger RNA sequences in a cell population, organ, or tissue sample, is a powerful means of generating comprehensive genome-level data sets on complex diseases. We have recently applied a systematic transcriptome-based ...

For many years, our laboratory has been developing gene transfer approaches for salivary gland disorders that currently lack effective therapy. The purpose of this chapter is to describe key methods used in this developmental process. Specifically, we focus on one clinical condition, i ...

Escherichia coli O157:H7, a major cause of food-borne illness, produces Shiga toxins (Stxs) that block protein synthesis by inactivating the ribosome. In this chapter, we describe a simple cell-based fluorescent assay to detect Stxs and inhibitors of toxin activity. The assay can also be used ...

The ability to rapidly differentiate Bacillus anthracis spores from spores belonging to other Bacillus spp. is potentially useful for combating the intentional release of this biothreat agent. Furthermore, not all B. anthracis strains are fully virulent and the ability to determine t ...

Cyanobacterial mass occurrences are widespread and often contain hepatotoxic, i.e. microcystin- and nodularin-producing, species. Nowadays, detection of microcystin (mcy) and nodularin synthetase (nda) genes is widely used for the recognition of toxic cyanobacterial stra ...

Shiga toxin-producing Escherichia coli is a leading cause of human gastroenteritis from food and waterborne sources worldwide. Shiga toxins 1 and 2 are important virulence factors linked to severe human illness. In particular, Shiga toxin 2 is composed of a diverse and heterogeneous group ...

Bacterial endotoxins or lipopolysaccharides (LPS) are among the most potent activators of innate immune system, yet mechanisms of their action and, in particular, the role of the glycans remain elusive. Efficient noninvasive labeling strategies are necessary for studying intera ...

Treatment of infections caused by Gram-negative bacteria is difficult due in large part to problems arising from innate and acquired drug resistance, resulting in a limited number of effective antibiotics. Consequently, antibiotics that can circumvent mechanisms of drug resista ...

Lipopolysaccharides (LPSs) are major components of the external membrane of Gram-negative bacteria, and act as an effective permeability barrier. They are essentially composed of a hydrophilic polysaccharide region linked to an hydrophobic one, termed lipid A. Depending on their i ...

This chapter presents a conjugation method for coupling probes bearing hydrazine or primary amino groups to a smooth(S)-form lipopolysaccharide (LPS). LPS is modified by the activation of the hydroxyl groups present in its O-antigen moiety with cyanogen bromide in aqueous acetone. The m ...

This chapter presents a conjugation method for coupling probes bearing hydrazine or primary amino groups to a lipopolysaccharide (LPS). LPS is modified by the activation of the hydroxyl groups present in its O-antigen moiety with 1-cyano-4-dimethylaminopyridinium tetrafluoro ...

Polyacrylamide slab gel electrophoresis in the presence of sodium dodecyl sulfate or sodium deoxycholate (SDS- or DOC-slab-PAGE) is a powerful technique for the separation of smooth(S)-type bacterial lipopolysaccharides (LPS). In order to recover the individual LPS species from t ...

Endotoxins (lipopolysaccharides, LPSs) are components of the envelope of Gram-negative bacteria. These molecules, responsible for both advantageous and harmful biological activities of these microorganisms, are highly immunogenic and directly involved in numerous bac ...