The Application of PCR Fingerprinting to the Epidemiologic Analysis of Bacterial and Fungal Pathogens
The increased incidence of nosocomial infections especially in immunocompromised patients has stimulated interest in developing more definite procedures for epidemiological studies of the etiological agents, such as pathogenic strains of different bacterial and fungal species. To determine the origins of infection, the routes of acquisition and transmission as well as the persistence of pathologic strains, precise and reproducible diagnostic methods are required. Recently, Williams et al (1990) and Welsh and McClelland (1990) described a PCR-based method for assessing DNA polymorphisms by amplifying genomic DNA with single primers of arbitrary nucleotide sequence. The use of the PCR with primers differing in length and nucleotide composition detected polymorphisms in the absence of specific sequence information in DNA from bacteria (Jayarao et al 1992), fungi (Crowhurst et al 1991), plants (Wilde et al 1992), animals (Welsh et al 1991) and humans (Williams et al 1990). Polymorphisms generated by this method have been termed arbitrarily primed-polymerase chain reaction (AP-PCR) fingerprinting (Welsh and McClelland 1990), random amplification of polymorphic DNA (RAPD) markers (Williams et al 1990) and DNA amplification fingerprinting (Jayarao et al 1992). The term PCR fingerprinting is used here because single primers, which were originally applied as hybridization probes in conventional DNA fingerprinting to detect minisatellite and microsatellite DNA such as the core sequence of phage M 13 (5’GAGGGTGGCGGTTCT), and the simple repeat sequences (GACA)4 and (GATA)4 (Meyer et al.1991; Ali et al 1986) were used to amplify DNA sequences in the genome of different bacterial and yeast species. Besides, in our experiments primers were tested which were already used in other labs for AP-PCR (universal sequence of M 13; 5’TTATGAAACGACGGCCAGT; Welsh et al 1991) or RAPD (10-mer oligonucleotide; 5’TCACGATGCA; Williams et al).
KeywordsMagnesium Phenol Hydroxide Agarose Bromide
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