ABSTRACT
Until recently, morphotyping, a method evaluating fringe and surface characteristics of streak colonies grown on malt agar, has been recommended as a simple and unexpensive typing method for Candida albicans isolates. The discriminatory power and reproducibility of Hunter's modified scheme of Phongpaichit's morphotyping has been evaluated on 28 C. albicans isolates recovered from the oral cavity of asymptomatic human immunodeficiency virus-positive subjects, and compared to two molecular typing methods: randomly amplified polymorphic DNA (RAPD) fingerprinting, and contour clamped homogeneous electric field (CHEF) electrophoretic karyotyping. Morphological features of streak colonies allowed to distinguish 11 different morphotypes while RAPD fingerprinting yielded 25 different patterns and CHEF electrophoresis recognized 9 karyotypes. The discriminatory power calculated with the formula of Hunter and Gaston was 0.780 for morphotyping, 0.984 for RAPD fingerprinting, and 0.630 for karyotyping. Reproducibility was tested using 43 serial isolates from 15 subjects (2 to 6 isolates per subject) and by repeating the test after one year storage of the isolates. While genetic methods generally recognized a single type for all serial isolates from each of the subjects studied, morphotyping detected strain variations in five subjects in the absence of genetic confirmation. Poor reproducibility was demonstrated repeating morphotyping after one year storage of the isolates since differences in at least one character were detected in 92.9 percent of the strains.
Subject(s)
Humans , AIDS-Related Opportunistic Infections/microbiology , Candida albicans/classification , Candidiasis, Oral/microbiology , Mycological Typing Techniques/methods , Candida albicans/genetics , Candida albicans/isolation & purification , Genetic Techniques , Reproducibility of ResultsABSTRACT
BACKGROUND & OBJECTIVES: The heterogeneity of group D streptococci led to the identification of various biotypes of Streptococcus equinus and Streptococcus bovis and to the description of new species. The objective of the present study was to improve the phenotypic delineation between species and to clarify their respective phylogenetic position. METHODS: Physiological and genomic analyses were carried out in 84 representative strains of the group D streptococci. Biotypes were determined with the API 20 strep and rapid ID 32 STREP systems of identification. Quantitative DNA-DNA hybridization under stringent conditions and values of the deltaT(m) allowed to delineate species and subspecies. The phylogenic position of the different genomic groups was determined by comparing the sequences of their 16S rDNA. RESULTS: Four DNA-clusters, including seven species or subspecies, were characterized. Differential associations of biochemical characters allowed their identification. S. equinus and the type strain of S. bovis belonged to a single species. S. gallolyticus, S. bovis biotype II.2, and S. macedonicus formed a single DNA-cluster including three different subspecies. These were designated as S. gallolyticus subsp. gallolyticus, S. gallolyticus subsp. pasteurianus, and S. gallolyticus subsp. macedonicus. The two other DNA-clusters corresponded to the two subspecies of S. infantarius, and to S. alactolyticus. INTERPRETATION & CONCLUSION: This study presented a new classification associated with an identification scheme of group D streptococci. The changes in this classification demonstrate the interest of a polyphagic approach of the bacterial identification.