Brucellosis 2003 proceedings - PHIDIAS

Poster Session
104- IDENTIFICATION OF SPECIES OF Brucella USING FOURIER TRANSFORM
INFRARED SPECTROSCOPY (FTIR).
M. A. Miguel Gómez 1 , M. A. Bratos Pérez 1 , F. J. Martín Gil 2 , A. Dueñas Díez 1 , J. F. Martín Rodríguez 3 ,
P. Gutiérrez Rodríguez 1 , A. Orduña Domingo 1 , A. Rodríguez Torres 1 . (1) Departamento de
Microbiología, Hospital Clínico Universitario, Facultad de Medicina. Valladolid, Spain. (2) Servicio de
Análisis Clínicos. Hospital Universitario Río Hortega, Spain. (3) Departamento de Estadística,
Facultad de Medicina, Valladolid, Spain
Fourier Transform Infrared Spectroscopy is a technique that has been used
over the years in chemical analysis for the identification of substances and is one that
may be applied to the characterisation of microorganisms. Since 80s it has
successfully been applied to the differentiation of a wide range of microorganisms.
Bacteria belonging to the Brucella genus are classified into six recognised species B.
melitensis, B. abortus, B. suis, B. canis, B. ovis and B. neotomae, within some of
which different biovars have been recorded. The marked tendency towards variation
in the smooth rough phase, together with the laboriousness and risk involved in the
methods used in their identification make their classification difficult. We studied the
type strains of the different species and biovars of Brucella and 11 isolates of human
origin of B. melitensis, six corresponding to biovar 1, one to biovar 2 and five to
biovar 3. The results of lineal discriminant analysis performed using the data provide
an above 95% likelihood of correct classification, over half of which are in fact above
99% for the vast majority of Brucella strains. Only one case of B. melitensis biovar 1
has been incorrectly classified. The rest of the microorganisms studied have been
classified correctly in all cases to a likelihood of over 80%. In the graphic
representation of the analysis a grouping of these can be seen in clusters, which
include the different species. One of these comprises B. melitensis, another B.
abortus, and another wider one is made up of B. suis. The B canis, B. ovis and B.
neotomae strains appear separate from the previously described groups.
105- AFLP: A TOOL FOR THE IDENTIFICATION AND TYPING OF Brucella
ISOLATES?.
Adrian M. Whatmore, Terry Murphy, Sally J. Cutler, and Alastair P. Macmillan. Department of
Statutory and Exotic Bacterial Diseases, Veterinary Laboratories Agency, Addlestone, Surrey, KT15
3NB, United Kingdom.
Brucellosis is a zoonotic disease of major public health, animal welfare and
economic significance worldwide. Although the UK has achieved Officially
Brucellosis-Free (OBF) status the constant threat of reintroduction highlights the
need for improved diagnostic and epidemiological tools. Such tools enhance the
speed of management of outbreaks by facilitating rapid identification of the infecting
organism, its source and means of spread. In light of the complexity of classical
Brucella typing the application of molecular techniques provides new approaches that
will speed up the identification and typing process and hence reduce the risk of
spread and the threat to human and animal welfare. Here we describe the
development of AFLP procedures applicable to Brucella. Some fifty distinct
combinations of restriction enzymes and/or selective primers were tested. Two, which
gave the most promising profiles in terms of band size range, band spread and band
number and showed differences between reference strains of distinct Brucella
species, were selected for further study. These combinations (EcoRI+0/MseI+TC and
Brucellosis 2003 International Research Conference
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