实验基础

Human enteroviruses include polioviruses, coxsackieviruses, echoviruses, and newer enteroviruses designated enteroviruses 68–71. They are classified as one genus of the Picornaviridae; rhinoviruses are a separate genus of the same virus family. All picornaviruses are sma ...

Infants and young children undergoing their primary infection with common human respiratory viruses are at risk of serious, even life-threatening, lower respiratory tract infection, A multiplicity of viruses infect the human respiratory tract but a relatively small number are res ...

Poxviruses are large, double-stranded DNA viruses that replicate in the cytoplasm of infected cells. The family Poxviridae is divided into two subfamilies, the Entomopoxvirinae of insects and the Chordopoxvirinae of vertebrates; the latter consists of eight genera and several unc ...

Rotavirus has been recognized as the major etiological agent of acute gastroenteritis in infants and young children. The rotaviruses contain a genome of 11 segments of double-stranded RNA that can be separated into distinct bands by electrophoresis. The migration pattern of the 11 genome ...

The principle of time-resolved fluorometry and its application to solidphase immunoassays (TR-FIAs) was invented in the 1980s in Turku, Finland at the Wallac Biochemical Laboratory and University of Turku (1,2). One of the early diagnostic applications was the detection of viral antig ...

Sandfly fever Sicilian (SFS) virus (genus Phlebovirus, family Bunyaviridae) is a member of the sandfly fever virus serogroup, which also includes sandfly fever Naples and Toscana viruses. These viruses are transmitted by Phlebotomus sandflies in the Mediterranean and the Middle Eas ...

In 1953, the first adenovirus was isolated from a human and, subsequently, 47 types have been shown to exist. Adenoviruses are now classified into six subgroups (A–F), which are based on their hemagglutination properties (1). They have a icosahedron structure that contains double-stranded l ...

Sensitive immunoassays were developed in the 1950s and 1960s using radioactive isotopes. The application of enzymes as labels (1,2) in the late 1960s increased the potential sensitivity and safety of immunoassays. The widespread popularity of enzyme-linked immunosorbent assays ...

The improvements in DNA-sequencing technology in recent years has opened new possibilities, from large scale genome projects to routine diagnostic applications. Among the most important developments towards automated systems has been the introduction of the polymerase chain ...

The environment furnishes a variety of surroundings that may favor the survival of microorganisms. The aquatic environment (which includes marine and fresh water, raw and treated sewage, sludge, and sediments) often provides conditions in which pathogenic viruses released from the ...

In general, three assay types are commonly used in diagnostic virology. Since these are described in other chapters it suffices here just to reiterate the methodology names: 1.

Traditionally, the accurate detection of viruses and the diagnosis of viral diseases has been difficult and expensive as viruses cannot be visualized by conventional light microscopy and need propagation in primary or continuous cell culture. The advent and increasing sophistica ...

During the past 30 years, several strategies have been used to synthesize DNA. Moreover, it was not possible to utilize automated DNA synthesis until the phosphate triester method using phosphoramidite reagents was introduced (1,2). Since then, chemically synthesized oligonucleo ...

Nucleic acid sequence-based amplification (NASBA) is a primer dependent, homogeneous, isothermal amplification process for the detection of RNA (1). It can result in 109- fold amplification of a specific RNA sequence and offers the unique possibility to amplify RNA in a background of geno ...

An issue of some biological currency pertains to the identification of the functional potential of cells as deduced from the nucleotide sequence of their genes. In effect, nucleotide sequences of genes assigned in some confidence to expressed proteins and function in one cell are taken, when ...

The fluorescence stains used most often for mycoplasma detection are DNA binding fluorochromes (DNAFs) and fluoresceinated antibodies. Both permit direct visualization of individual mycoplasma organisms. DNAFs will bind to any appropriately conformed DNA that is present in a s ...

The lack of a cell wall means that the growth of mycoplasmas can be inhibited by specific antiserum and provides the basis of a simple, economic, and objective means of species identification. In the diagnostic laboratory, unknown mycoplasma isolates are inoculated onto solid media and exami ...

β-lactamases confer resistance to β-lactam antibiotics, which are the most widely used family of antibiotics. It is, therefore, essential that one can identify the production of β-lactamases by clinical isolates and have effective ways of distinguishing the different enzymes. This is n ...

A significant proportion of microbial ecology is now based on the description of community structure in naturally occurring bacterial assemblages. The development of molecular biological techniques has facilitated this task, primarily via the cloning and sequencing of microb ...

Since the 1980s the use of ribosomal RNA (rRNA) sequence-based analysis to characterize microbial populations (mainly bacterial and archaeal populations) has increased significantly. This increased use is in response to the recognition that culture-based methods grossly mis ...