A new DNA probe assay (INNO LiPA Mycobacteria; Innogenetics, Ghent, Belgium) for the simultaneous identification, by means of reverse hybridization and line-probe technology, of Mycobacterium tuberculosis complex, Mycobacterium kansasii, Mycobacterium xenopi, Mycobacterium gordonae, the species of the Mycobacterium avium complex (MAC), Mycobacterium scrofulaceum, and Mycobacterium chelonae was evaluated on a panel of 238 strains including, besides representatives of all the taxa identifiable by the system, a number of other mycobacteria, some of which are known to be problematic with the only other commercial DNA probe system (AccuProbe; Gen-Probe, San Diego, Calif.), and two nocardiae. The new kit, which includes a control probe reacting with the whole genus Mycobacterium, correctly identified 99.6% of the strains tested; the one discrepancy, which remained unresolved, concerned an isolate identified as MAC intermediate by INNO LiPA Mycobacteria and as Mycobacterium intracellulare by AccuProbe. In five cases, because of an imperfect checking of hybridization temperature, a very slight, nonspecific, line was visible which was no longer evident when the test was repeated. Two strains whose DNA failed amplification at the first attempt were regularly identified when the test was repeated. Interestingly, the novel kit dodged all the pitfalls presented by the strains giving anomalous reactions with AccuProbe. A unique feature of INNO LiPA Mycobacteria is its ability to recognize different subgroups within the species M. kansasii and M. chelonae, while the declared overlapping reactivity of probe 4 with some M. kansasii and Mycobacterium gastri organisms and of probe 9 with MAC, Mycobacterium haemophilum, and Mycobacterium malmoense, may furnish a useful aid for their identification. The turnaround time of the method is approximately 6 h, including a preliminary PCR amplification.

The observation of microbial contaminations in a time close to the transplant (i.e., at the end of the preservation period and in the deswelling step) showed that a fast microbial tests during the deswelling procedure may prevent the grafting of a contaminated cornea. The appearance of bacteria in the deswelling medium despite a negative culture medium suggests that bacteria penetrate the corneal tissues during the culture to be subsequently extruded when the internal fluids move outward.

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INTRODUZIONELe innovazioni tecnologiche, il profilarsi sempre più evidente di quella che diverrà un'emergenza infettiva per le nostre popolazioni, la globalizzazione e l'impatto importante dell'antropizzazione sulla biosfera, stanno cambiando profondamente lo scenario in cui si trovano a lavorare i microbiologi e comportando modifiche tali da porre seriamente il problema di prepararsi ed attrezzar- Another critical point is represented by the possible insurgence of infections due to agents used in bioterrorism.More than ever it is indispensable for our country to define a network of structures able to give an organized response to these relevant problems both in terms of prevention and assistance. It is therefore important to define a protocol for clinical microbiology on a national basis that responds to different levels of activity. Keeping in mind that the fundamental objectives of microbiology are: -diagnosis of infective diseases -determination of resistance to antimicrobic drugs as a valid instrument to allow the clinician to proceed with specific treatment -control of infection spreading in the sanitary structures (hospital infections) -to develop front line defence in new infections and bioterrorism, in collaboration with the department of prevention -to be a reference centre for specific trainingThe principals on which one must base the reorganization are essentially three: 1. to configurate the microbiology service on the basis of real needs of the local population 2. to give qualitatively optimal results in real time 3. supporting "good clinical practice" to assure adequate patient results and acceptable costs for the regionale sanitary system (SSR) The organization of microbiology should therefore foresee a microbiology department, structured on a provincial basis or a vast area including a structured centralized complex with peripheral structures (even simplified). This must integrate actively with the department of prevention in regard to the dynamics of infectious diseases in the territory and with the Division of Infectious Diseases (where this exists). In parallel an active interaction with research structures (IRCCS and University) present in the territory to enable the coordinated implementation of the guidelines of research and its relapse on diagnostic exams. Keeping in mind that the quality of diagnostic performance is closely linked to the professional experience of the operators it is good practice centralizing some diagnostic procedures by identifying a reference centre. In a regional network organized in this way it is important to include a coordinating structure, since it is necessary to activate a complete level of communication, information exchange, activity planning, and complete problem solving.

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