Clinical Microbiology and Infection最新文献

Objectives: The CHAGASAZOL trial compared posaconazole and benznidazole for treating chronic Chagas Disease. Posaconazole showed poor short-term efficacy by means of qPCR compared to benznidazole, but few studies have reported long-term follow-up using this tool. The aim of the present study is to provide a more comprehensive analysis of the CHAGASAZOL cohort through 11 years of follow-up.

Methods: This is a prospective observational cohort of individuals who were included in the CHAGASAZOL trial. Data was censored as of December 31, 2023. Subjects initially treated with posaconazole with a positive qPCR were offered re-treatment with Benznidazole. All patients underwent clinical and electrocardiographic evaluations as well as a qPCR at a six-month or one-year interval. The primary objective was parasitological failure, defined as any positive qPCR in peripheral blood at any time during follow-up.

Results: Seventy-two participants were enrolled (median follow-up: 71 months, range 1-147). At baseline, 59 (82%) were classified as indeterminate form, 9 (12%) as cardiac, 2 (3%) as digestive and 2 (3%) as mixed forms. Forty-eight participants received posaconazole, 45 completing at least one follow-up visit. Up to 43/45 (95%) presented a positive qPCR, and of them, 35 accepted to be retreated with benznidazole. Considering those treated with benznidazole (either initially or as a re-treatment), only 3/51 (6%) showed a positive qPCR. Four (5.5%) participants showed cardiac progression after 3 to 10 years of follow-up, with an incident rate of 0.94 events per 100 person-years. Two of them had received the complete benznidazole treatment, 1 was partially treated (17 days) and 1 was only treated with posaconazole before clinical progression.

Conclusion: Even if benznidazole showed parasitological efficacy, lifelong follow-up should be offered to individuals living with CD, as both parasitological failure and clinical progression can occur many years after diagnosis and treatment.

Objectives: Treponema pallidum subsp. pallidum (T. pallidum) is the etiological agent of syphilis, a sexually transmitted disease of global public health importance. The objective of this study was to introduce a novel in vitro protocol for isolation of T. pallidum directly from patients' clinical samples, eliminating the need for rabbit propagation.

Methods: Four oral and five genital swabs were collected from nine epidemiologically unrelated patients at two hospitals in Brno, Czech Republic. Swabs were submerged in TpCM-2 medium for transport. Then, samples were placed on a 0.4 μm filters and incubated for 2.5 hours. During this period, spiral T. pallidum cells passed through the filter pores to the well containing TpCM-2 medium and rabbit feeder cells (Sf1Ep). Stable T. pallidum cultures (containing >1 × 10⁷ treponemes) were achieved through subculturing every 7 days into fresh well.

Results: A successful protocol for in vitro isolation of T. pallidum was established. From the nine clinical specimens processed, six T. pallidum cultures (MU1-MU6) were derived after 14-112 days of cultivation. Five of these strains (MU1-MU5) belonged to SS14-like cluster and shared the same allelic profile 1.3.1. The remaining strain (MU6) was identified as a Nichols-like strain with an allelic profile 9.16.3.

Conclusions: The introduced in vitro protocol enables isolation of T. pallidum from clinical material, including frozen samples, without the need for experimental rabbits. This method facilitates the isolation of contemporary, clinically relevant treponemal strains.