Mortality related to candidemia and risk factors associated with non-candida albicans

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Isolation and drug susceptibility of Candida parapsilosis sensu lato and other species of C. parapsilosis complex from patients with blood stream infections and proposal of a novel LAMP identification method for the species

Candida parapsilosis complex (CPC) is the third Candida species isolated in blood cultures of patients from our Hospital, following C. albicans and C. tropicalis. From 2006 to 2010, the median annual distribution of CPC was 8 cases/year. Records of 36 patients were reviewed. CPC were 31 (86.1 %) C. parapsilosis; 4 (11.1 %) C. orthopsilosis; and 1 (2.8 %) C. metapsilosis. Clinical characteristics were central venous catheter, 34 (94.4 %); parental nutrition, 25 (70 %); surgery, 27 (57.9 %); prior bacteremia, 20 (51.3 %); malignancy, 18 (50 %). General mortality was 47.2 %. Death was higher in immunosuppressed patients (17 vs. 11; p = 0.003). Three out four (75 %) patients with C. orthopsilosis and 14 out 31 (45.2 %) with C. parapsilosis died (p = 0.558). Thirty-nine individual isolates were tested for susceptibility to seven antifungal drugs, with MICs values showing susceptibility to all of them. Two isolates, one C. orthopsilosis and one C. parapsilosis, had fluconazole MIC = 4 mu g/mL. Differentiation among CPC has implication in caring for patients with invasive candidiasis since there are differences in virulence, pathogenicity and drug susceptibility. A method targeting the topoisomerase II gene based on loop-mediated isothermal amplification (LAMP) was developed. LAMP emerges as a promising tool for the identification of fungal species due to the high sensitivity and specificity. LAMP can be performed at the point-of-care, being no necessary the use of expensive equipment. In our study, the method was successful comparing to the DNA sequencing and proved to be a reliable and fast assay to distinguish the three species of CPC1791-25362sem informaçã

Comparison of DNA microarray, Loop-Mediated Isothermal Amplification (LAMP) and real-time PCR with DNA sequencing for identification of Fusarium spp. obtained from patients with hematologic malignancies

The performance of three molecular biology techniques, i.e., DNA microarray, loop-mediated isothermal amplification (LAMP), and real-time PCR were compared with DNA sequencing for properly identification of 20 isolates of Fusarium spp. obtained from blood stream as etiologic agent of invasive infections in patients with hematologic malignancies. DNA microarray, LAMP and real-time PCR identified 16 (80%) out of 20 samples as Fusarium solani species complex (FSSC) and four (20%) as Fusarium spp. The agreement among the techniques was 100%. LAMP exhibited 100% specificity, while DNA microarray, LAMP and real-time PCR showed 100% sensitivity. The three techniques had 100% agreement with DNA sequencing. Sixteen isolates were identified as FSSC by sequencing, being five Fusarium keratoplasticum, nine Fusarium petroliphilum and two Fusarium solani. On the other hand, sequencing identified four isolates as Fusarium non-solani species complex (FNSSC), being three isolates as Fusarium napiforme and one isolate as Fusarium oxysporum. Finally, LAMP proved to be faster and more accessible than DNA microarray and real-time PCR, since it does not require a thermocycler. Therefore, LAMP signalizes as emerging and promising methodology to be used in routine identification of Fusarium spp. among cases of invasive fungal infections1827-8625632FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2011/16205-5Japan Science & Technology Agency (JST); SATREPS; Ministry of Education, Culture, Sports, Science and Technology, Japan (MEXT); Japan Society for the Promotion of Science; Grants-in-Aid for Scientific Research (KAKENHI

Airborne transmission of invasive fusariosis in patients with hematologic malignancies

<div><p>From 2006 to 2013, an increasing incidence of fusariosis was observed in the hematologic patients of our University Hospital. We suspected of an environmental source, and the indoor hospital air was investigated as a potential source of the fungemia. Air samplings were performed in the hematology and bone marrow transplant (BMT) wards using an air sampler with pre-defined air volumes. To study the molecular relationship among environmental and clinical isolates, 18 <i>Fusarium</i> spp. recovered from blood cultures were included in the study. DNA sequencing of a partial portion of <i>TEF1α</i> gene was performed for molecular identification. Molecular typing was carried out by multi-locus sequence typing (MLST) using a four-gene scheme: <i>TEF1α</i>, rDNA, <i>RPB1</i> and <i>RPB2</i>. One hundred four isolates were recovered from the air of the hematology (n = 76) and the BMT (n = 28) wards. <i>Fusarium</i> isolates from the air were from five species complexes: <i>Fusarium fujikuroi</i> (FFSC, n = 56), <i>Fusarium incarnatum-equiseti</i> (FIESC, n = 24), <i>Fusarium solani</i> (FSSC, n = 13), <i>Fusarium chlamydosporum</i> (FCSC, n = 10), and <i>Fusarium oxysporum</i> (FOSC, n = 1). Fifteen <i>Fusarium</i> isolates recovered from blood belonged to FSSC, and three to FFSC. MLST identified the same sequence type (ST) in clinical and environmental isolates. ST1 was found in 5 isolates from blood and in 7 from the air, both identified as FSSC (<i>Fusarium petroliphilum</i>). STn1 was found in one isolate from blood and in one from the air, both identified as FFSC (<i>Fusarium napiforme</i>). <i>F</i>. <i>napiforme</i> was isolated from the air of the hospital room of the patient with fungemia due to <i>F</i>. <i>napiforme</i>. These findings suggested a possible clonal origin of the <i>Fusarium</i> spp. recovered from air and bloodcultures. In conclusion, our study found a diversity of <i>Fusarium</i> species in the air of our hospital, and a possible role of the air as source of systemic fusariosis in our immunocompromised patients.</p></div

Primers used for sequencing of clinical and environmental <i>Fusarium</i> isolates.

<p>Primers used for sequencing of clinical and environmental <i>Fusarium</i> isolates.</p

Distribution of <i>Fusarium</i> species isolated from hospital air.

<p>The frequency of each species complex in the hematology (A, n = 76) and BMT (B, n = 28) wards is shown. The species identified for each complex is presented outside the graphs. The species found exclusively in hematology unit are marked with (*). FCSC: <i>F</i>. <i>chlamydosporum</i> species complex; FFSC: <i>F</i>. <i>fujikuroi</i> species complex; FIESC: <i>F</i>. <i>incarnatum-equiseti</i> species complex; FOSC: <i>F</i>. <i>oxysporum</i> species complex; FSSC: <i>F</i>. <i>solani</i> species complex.</p