Ocular tuberculosis has traditionally been considered uncommon or anecdotal. Imprecise and variable diagnostic criteria have contributed to the confusion surrounding this topic. The increase in extrapulmonary manifestations of tuberculosis during the AIDS era established the need for a prospective study of ocular involvement in patients with all types of tuberculosis using well-defined criteria. During a 15-month period, 300 cases had culture-proven tuberculosis at our institution. We randomly selected 100 for systematic ophthalmologic evaluation. Our criteria for ocular tuberculosis were divided as follows: certainty (isolation of Mycobacterium tuberculosis from ocular specimens), probability (patients with isolation of M. tuberculosis from extraocular samples, with ocular lesions not attributable to other causes that respond to anti-tuberculous treatment), and possibility (same as probability but follow-up impossible). Ocular tuberculosis was present in 18 patients (18%) of which 10 patients fulfilled probability and 8 patients fulfilled possibility criteria. Eleven of 18 patients had HIV infection. In 11 patients, ocular involvement was asymptomatic. Almost all patients (17/18) had choroiditis, and other ocular lesions included papillitis, retinitis, vitritis, vasculitis, dacryoadenitis, and scleritis. Multivariate analysis showed as risk factors independently predicting ocular involvement in patients with ocular tuberculosis the presence of miliary disease (odd ratio 43.92, p = 0.002), ocular symptoms (odds ratio 6.35, p = 0.0143), and decreased visual acuity (odds ratio 0.04, p = 0.012). We observed an unexpectedly high (18%) incidence of ocular involvement, frequently asymptomatic, in patients with tuberculosis. Miliary disease is a clear predisposing factor in both HIV-infected and noninfected populations. Ocular examination should be routinely considered in patients with proven or suspected tuberculosis.

In recent years, matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) has proved a rapid and reliable method for the identification of bacteria and yeasts that have already been isolated. The objective of this study was to evaluate this technology as a routine method for the identification of microorganisms directly from blood culture bottles (BCBs), before isolation, in a large collection of samples. For this purpose, 1000 positive BCBs containing 1085 microorganisms have been analysed by conventional phenotypic methods and by MALDI-TOF MS. Discrepancies have been resolved using molecular methods: the amplification and sequencing of the 16S rRNA gene or the Superoxide Dismutase gene (sodA) for streptococcal isolates. MALDI-TOF predicted a species- or genus-level identification of 81.4% of the analysed microorganisms. The analysis by episode yielded a complete identification of 814 out of 1000 analysed episodes (81.4%). MALDI-TOF identification is available for clinicians within hours of a working shift, as oppose to 18 h later when conventional identification methods are performed. Moreover, although further improvement of sample preparation for polymicrobial BCBs is required, the identification of more than one pathogen in the same BCB provides a valuable indication of unexpected pathogens when their presence may remain undetected in Gram staining. Implementation of MALDI-TOF identification directly from the BCB provides a rapid and reliable identification of the causal pathogen within hours.

We prospectively determined the antifungal susceptibility of yeast isolates causing fungemia using the Etest on direct blood samples (195 prospectively collected and 133 laboratory prepared). We compared the Etest direct (24 h of incubation) with CLSI M27-A3 and the standard Etest methodologies for fluconazole, voriconazole, posaconazole, isavuconazole, caspofungin, and amphotericin B. Strains were classified as susceptible, resistant, or nonsusceptible using CLSI breakpoints (voriconazole breakpoints were used for posaconazole and isavuconazole). Categorical errors between Etest direct and CLSI M27-A3 for azoles were mostly minor. No errors were detected for caspofungin, and high percentages of major errors were detected for amphotericin B. For the azoles, false susceptibility (very major errors) was found in only two (0.6%) isolates (Candida tropicalis and C. glabrata). False resistance (major errors) was detected in 46 (14%) isolates for the three azoles (in 23 [7%] after excluding posaconazole). Etest direct of posaconazole yielded a higher number of major errors than the remaining azoles, especially for C. glabrata, Candida spp., and other yeasts. Excluding C. glabrata, Candida spp., and other yeasts, the remaining species did not yield major errors. Etest direct for fluconazole, voriconazole, isavuconazole, and caspofungin shows potential as an alternative to the CLSI M27-A3 procedure for performing rapid antifungal susceptibility tests on yeast isolates from patients with fungemia. Etest direct is a useful tool to screen for the presence of azole-resistant and caspofunginnonsusceptible strains.