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DNA fingerprinting patterns of Candida species using HinfI endonuclease.

Strain delineation was performed by means of restriction endonuclease analysis (REA) of genomic DNA with the restriction enzyme HinfI followed by conventional electrophoresis. A total of 337 yeast isolates representing 21 Candida species and five non-Candida yeast species was evaluated. A survey of isolates showed that Candida albicans and non-albicans species could be divided into mutually exclusive groups, and that subgroups could be created. Individual REA patterns for 111 C. albicans isolates, four Candida krusei isolates and 35 Candida glabrata isolates varied greatly, whereas 11 Candida dubliniensis isolates, 48 Candida tropicalis isolates and 41 Candida guilliermondii isolates could be divided into two, nine and nine groups, respectively. REA of the 49 Candida parapsilosis isolates with HinfI, however, showed that 47 (95 %) of them belonged to one group. REA patterns of the other yeast isolates, representing 19 species, were also quite different at the species level. These results showed that REA with HinfI may be useful for the identification and strain delineation of common and emerging Candida species.

Candida↗

Detection of seven Candida species using the Light-Cycler system.

Due to the limitations of classical methods for the detection of systemic fungal infections and the high mortality rates associated with these infections, it has become essential to develop a quick, sensitive and specific detection assay. By using the Idaho Technologies Light-Cycler system, a qualitative real-time PCR system has been developed for the detection of the leading causes of systemic infection within the genus Candida. The sensitivity of the assay was comparable to previously described PCR methods (1-5 c.f.u. ml(-1)) and, by the use of a single Candida probe, it was able to detect, but not differentiate between, seven species of Candida (Candida albicans, Candida dubliniensis, Candida glabrata, Candida kefyr, Candida krusei, Candida parapsilosis and Candida tropicalis). Single-round amplification on the Light-Cycler allowed rapid turn-around of clinical samples (within one working day) and it was shown to be more sensitive than classical procedures, exposing 39 possible systemic infections that were not detected by blood culture.

Candida↗

Antibody response to Candida and its use in clinical practice.

Reference values for specific IgM, IgA and IgG antibodies against three defined Candida antigens were determined in 280 healthy Swedish children aged 1 month to 15 years. The antibody response in 10 children with Candida infections was also determined. Precipitating IgG antibodies to a mannan-free Candida protein antigen were detected only in children with Candida infection. The haemagglutinating IgM antibody response in healthy children to a polysaccharide Candida antigen was weak. Adult levels of < or = 320 (95% confidence interval, CI) were not reached even in the oldest children (< or = 160). All children with Candida infections had elevated IgM titres. Adult levels of IgA antibodies to Candida mannan were reached at 10-12 years of age; at 13-15 years the titres were higher (< or = 1600) than in healthy adults (< or = 1000, 95% CI). All children with an acute Candida infection had elevated IgA titres. Our study shows that antibody testing may be used to diagnose systemic Candida infection in children and to follow the progression and resolution of systemic Candida infection with the rise and fall of antibody titres.

Adolescent↗

Relationship between salivary histatin 5 levels and Candida CFU counts in healthy elderly.

OBJECTIVES: Few epidemiological studies have confirmed the antifungal activity of histatin 5 in saliva against Candida species. The purpose of this study was to examine the relationship between concentrations of histatin 5 and the number of cultivable Candida in saliva samples from elderly. METHODS: Whole saliva samples were obtained from 124 elderly people, 65 years or older, living in a rural community. The concentrations of histatin 5 in saliva samples were determined by the enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody. Total colony-forming units (CFUs) were counted on a selective medium for Candida. Multiple linear regression analysis was performed to determine the independent contribution of explanatory variables to Candida CFUs using age, sex, histatin 5 concentration and type of denture prosthesis as independent variables. RESULTS: Saliva samples from 104 subjects (84%) were candidal colony-positive. The youngest group (65-69 years old) showed significantly smaller Candida CFU counts than those in the older group. The mean Candida CFU count of denture wearers was significantly higher than that of non-denture wearers. Significantly negative associations were found between Candida CFU counts and histatin 5 level in the oldest group (p < 0.05) and in the full-denture wearers (p < 0.01). Multiple linear regression analysis revealed that Candida CFU counts were mostly associated with type of dentures, followed by histatin 5 concentration. CONCLUSION: Possible activity of histatin 5 against Candida in whole saliva of elderly people was epidemiologically confirmed. The area covered by the prostheses was a strong factor associated with the Candida CFU count.

Age Factors↗

Candida-specific antibodies during experimental vaginal candidiasis in mice.

Protective host defense mechanisms against vaginal Candida albicans infections are poorly understood. Although cell-mediated immunity (CMI) is the predominant host defense mechanism against most mucosal Candida infections, the role of CMI against vaginal candidiasis is uncertain, both in humans and in an experimental mouse model. The role of humoral immunity is equally unclear. While clinical observations suggest a minimal role for antibodies against vaginal candidiasis, an experimental rat model has provided evidence for a protective role for Candida-specific immunoglobulin A (IgA) antibodies. Additionally, Candida vaccination-induced IgM and IgG3 antibodies are protective in a mouse model of vaginitis. In the present study, the role of infection-induced humoral immunity in protection against experimental vaginal candidiasis was evaluated through the quantification of Candida-specific IgA, IgG, and IgM antibodies in serum and vaginal lavage fluids of mice with primary and secondary (partially protected) infection. In naïve mice, total, but not Candida-specific, antibodies were detected in serum and lavage fluids, consistent with lack of yeast colonization in mice. In infected mice, Candida-specific IgA and IgG antibodies were induced in serum with anamnestic responses to secondary infection. In lavage fluid, while Candida-specific antibodies were detectable, concentrations were extremely low with no anamnestic responses in mice with secondary infection. The incorporation of alternative protocols-including infections in a different strain of mice, prolongation of primary infection prior to secondary challenge, use of different enzyme-linked immunosorbent assay capture antigens, and concentration of lavage fluid-did not enhance local Candida-specific antibody production or detection. Additionally, antibodies were not removed from lavage fluids by being bound to Candida during infection. Together, these data suggest that antibodies are not readily present in vaginal secretions of infected mice and thus have a limited natural protective role against infection.

Animals↗

Cloning of the RHO1 gene from Candida albicans and its regulation of beta-1,3-glucan synthesis.

The Saccharomyces cerevisiae RHO1 gene encodes a low-molecular-weight GTPase. One of its recently identified functions is the regulation of beta-1,3-glucan synthase, which synthesizes the main component of the fungal cell wall (J. Drgonova et al., Science 272:277-279, 1996; T. Mazur and W. Baginsky, J. Biol. Chem. 271:14604-14609, 1996; and H. Qadota et al., Science 272:279-281, 1996). From the opportunistic pathogenic fungus Candida albicans, we cloned the RHO1 gene by the PCR and cross-hybridization methods. Sequence analysis revealed that the Candida RHO1 gene has a 597-nucleotide region which encodes a putative 22.0-kDa peptide. The deduced amino acid sequence predicts that Candida albicans Rho1p is 82.9% identical to Saccharomyces Rho1p and contains all the domains conserved among Rho-type GTPases from other organisms. The Candida albicans RHO1 gene could rescue a S. cerevisiae strain containing a rho1 deletion. Furthermore, recombinant Candida albicans Rho1p could reactivate the beta-1,3-glucan synthesis activities of both C. albicans and S. cerevisiae membranes in which endogenous Rho1p had been depleted by Tergitol NP-40-NaCl treatment. Candida albicans Rho1p was copurified with the beta-1,3-glucan synthase putative catalytic subunit, Candida albicans Gsc1p, by product entrapment. Candida albicans Rho1p was shown to interact directly with Candida albicans Gsc1p in a ligand overlay assay and a cross-linking study. These results indicate that Candida albicans Rho1p acts in the same manner as Saccharomyces cerevisiae Rho1p to regulate beta-1,3-glucan synthesis.

Amino Acid Sequence↗

Diagnosis of invasive candidiasis by a dot immunobinding assay for Candida antigen detection.

A dot immunobinding assay which uses a polyclonal rabbit anti-Candida immunoglobulin G as the primary antibody and colloidal gold coated with goat anti-rabbit immunoglobulin G as the secondary antibody for the detection of Candida cytoplasmic antigens is described. It was able to detect as little as 1 ng of total Candida protein per ml when a cytoplasmic extract of Candida albicans was seeded into buffer and 10 ng/ml when the same extract was seeded into pooled human serum. Serial serum samples from four groups of patients were assayed for Candida antigen: (i) 22 patients with candidemia, (ii) 16 patients at high risk for invasive candidiasis, (iii) 3 patients with other deep mycoses, and (iv) 50 hospitalized patients at low risk for serious Candida infection. Of the 22 candidemic patients, 19 had invasive candidiasis and 3 had transient candidemia. Antigenemia was detected in 16 of the 19 patients with invasive candidiasis (including patients with C. albicans, Candida tropicalis, Candida glabrata, Candida krusei, and Candida parapsilosis) and in 4 of 16 patients at high risk for invasive candidiasis. There was no detectable antigen in 12 high-risk control patients, 3 patients with transient candidemia, 3 patients with other deep mycoses, and 50 relatively low-risk patients. The sensitivity for detecting invasive disease in candidemic patients and specificity for all patients studied were 84.2 and 94.4%, respectively. The positive predictive value was 80%; the negative predictive value was 95.7%. The sensitivity for neutropenic patients with invasive disease was 85.7%. This assay is rapid and accurate and appears to be useful in identifying candidemic patients with invasive candidiasis.

Antibodies, Fungal↗

Activities of micafungin against 315 invasive clinical isolates of fluconazole-resistant Candida spp.

Micafungin is a new echinocandin exhibiting broad-spectrum activity against Candida spp. The activity of the echinocandins against Candida species known to express intrinsic or acquired resistance to fluconazole is of interest. We determined the MICs of micafungin and caspofungin against 315 invasive clinical (bloodstream and other sterile-site) isolates of fluconazole-resistant Candida species obtained from geographically diverse medical centers between 2001 and 2004. MICs were determined using broth microdilution according to the CLSI reference method M27-A2. RPMI 1640 was used as the test medium, and we used the MIC endpoint of prominent growth reduction at 24 h. Among the 315 fluconazole-resistant Candida isolates, 146 (46%) were C. krusei, 110 (35%) were C. glabrata, 41 (13%) were C. albicans, and 18 (6%) were less frequently isolated species. Micafungin had good in vitro activity against all fluconazole-resistant Candida spp. tested; the MICs at which 50% (MIC(50)) and 90% (MIC(90)) of isolates were inhibited were 0.03 microg/ml and 0.06 microg/ml, respectively. All the fluconazole-resistant Candida spp. were inhibited at a micafungin MIC that was </=1 microg/ml. Among the most common fluconazole-resistant Candida spp. tested in the collection, C. glabrata exhibited the lowest micafungin MICs (MIC(90), </=0.015 microg/ml), followed by C. albicans (MIC(90), 0.03 microg/ml) and C. krusei (MIC(90), 0.06 microg/ml). The new echinocandin micafungin has excellent in vitro activity against 315 invasive clinical isolates of fluconazole-resistant Candida, which represents the largest collection to date of fluconazole-resistant Candida isolates tested against micafungin. Micafungin may prove useful in the treatment of infections due to azole-resistant Candida.

Antifungal Agents↗

Mechanisms of attachment of neutrophils to Candida albicans pseudohyphae in the absence of serum, and of subsequent damage to pseudohyphae by microbicidal processes of neutrophils in vitro.

Mechanisms were studied that might explain the attachment and damage to Candida albicans pseudohyphae by neutrophils in the absence of serum. Attachment of neutrophils to pseudo hyphae was inhibited by Candida mannans (1-10 mg/ml), but not by mannose, dextran, chitin, conconavalin A, or highly charged polyamino acids. Contact was also inhibited by pretreatment of Candida before incubation with neutrophils with chymotrypsin, but not trypsin or several inhibitors of proteases. Similar results were obtained with pretreatment of neutrophils, except that trypsin was inhibitory. When pseudohyphae were killed with ultraviolet light, proteinpolysaccharide complexes of mol wt <10,000 were released which appeared to bind to the surfaces of neutrophils and inhibit contact between neutrophils and Candida, as well as other fungi. Damage to Candida by neutrophils was inhibited by agents known to act on neutrophil oxidative microbicidal mechanisms, including sodium cyanide, sodium azide, catalase, superoxide dismutase, and 1, 4 diazobicyclo (2, 2, 2) octane, a singlet oxygen quencher. Neutrophils from a patient with chronic granulomatous disease did not damage Candida at all. However, the hydroxyl radical scavengers mannitol and benzoate were not inhibitory. Cationic proteins and lactoferrin also did not appear to play a major role in this system. Low concentrations of lysozyme which did not damage Candida in isotonic buffer solutions damaged pseudohyphae in distilled water. Isolated neutrophil granules damaged pseudohyphae only with added hydrogen peroxide and halide, and damage occurred only with granule fractions known to contain myeloperoxidase. These findings suggest that neutrophils recognized a molecule on the Candida surface which has a chymotrypsin sensitive protein component, and which may be liberated from the cell surface upon death of organism. The neutrophil receptors for Candida appear to be sensitive to trypsin and chymotrypsin. Damage to Candida by neutrophils occurred primarily by oxidative mechanisms, including the production of superoxide and hydrogen peroxide interacting with myeloperoxidase and halide, as well as singlet oxygen, but did not appear to involve hydroxyl radical. Lysozyme might have an accessory role, under some conditions.

Candida albicans↗

In vitro activity of 6 antifungal agents on candida species isolated as causative agents from vaginal and other clinical specimens.

OBJECTIVE: To study the susceptibility pattern of candida species isolated from various clinical specimens to common usable antifungals in Libya. METHODS: Two hundred and four candida species were isolated from patients complaining of fungal infections gathered from vaginal swabs, nails, throat, hair and ear. Yeast isolates were identified to the species level by API 20C AUX Commercial system. The in vitro susceptibility to amphotericin B, nystatin, ketoconazole, miconazole, clotrimazole and econazole was determined using the macrodilution in broth method. RESULTS: Candida albicans was the most common isolated species from vaginal and Candida tropicalis from throat swabs. On the other hand Candida parapsilosis and Candida guilliermondii were more often isolated from the ear and hair. Candida krusei and Candida fomata were only isolated from vulvaginitis in this study. In vitro sensitivity showed that most of the isolates were inhibited at optimum ranges of minimal inhibitory concentration particularly with amphotericin B and nyastatin. On the other hand resistance strains of candida species were found against the 4 azoles antifungal agents. CONCLUSION: Candida albicans and Torulopsis glabrata are among the most common cause of vaginitis among Libyan females. Amphotericin B, nystatin and clotrimazole were the most effective antifungals against most isolates in this study. Fungicidal effect was obtained with most antifungals at higher concentrations.

Antifungal Agents↗

[The distribution of Candida spp. in the oral cavities of normal children].

OBJECTIVE: To study the distribution of Candida spp. mainly Candida albicans in the oral cavities of health children. METHODS: Four groups children of different ages, A1: newborn babies, A2: 3.2 years old (average) children,A3, 7.2 years old,A4:12.7 years old and B control group 20.4 years old,mucosal swab sampling with centrification,CHROMagar Candida identified culture medium for culture and identification, and different methods for isolating Candida albicans for A3 group. RESULTS: The isolation rates of Candida spp were A1 7.5%,A2 70%,A3 56.36%,A4 49.12%,B 27.5%,the proportion of Candida albicans also diversely,and the method of PCR was more sensitive than the one of culture. CONCLUSION: Candida spp can be isolated from the normal oral cavities in different aged children, most of them were Candida albicans,both the isolation rates of Candida spp and the proportion of Candida albicans were different.

Adolescent↗

Trends in neonatal septicemia: emergence of non-albicans Candida.

In a prospective analysis, blood from 660 neonates admitted to neonatal Intensive Care Unit (NICU) of a teaching hospital with clinical suspicion of septicemia was cultured to look for etiological agents with particular reference to role of Candida species. Blood culture specimens from two different sites at same time were obtained to rule out possibility of a Candida isolate being a mere contaminant. Due to technical difficulties, this was possible in only 338 neonates (Group I); from remaining 322 neonates only single specimen was available (Group II). Candida was isolated from total 90 neonates (isolation rate 13.6%) and it was the single most common isolate. Majority were non-albicans Candida (germ tube test negative - 76/90). In group I, Candida was isolated from 66 neonates, of these 49 grew Candida in both specimens (significant candidemia). 44 records were available for analysis. Low birth weight was found in 73.3%. Crude mortality was 52.6%. A peak in isolation rate of Candida was noted (isolation rate 27%, p<0.05) in month of February. In Group I, 49 of the total 66 (74.2%) isolates of Candida were significant, suggesting that three in every four Candida isolated from blood can be significant. Non-albicans Candida are emerging as important pathogens for neonatal septicemia.

Candida↗

Phenotypic and genotypic identification of Candida spp. isolated from hospitalized patients.

As candidosis incidence continue to rise, quick laboratory identification of Candida species is becoming increasingly important for a growing population of patients at-risk. RAPD techniques were used on samples of Candida obtained from patients hospitalized at Santa Casa de Misericordia in Belo Horizonte (SCMBH) Brazil, from March 1998 to December 2000 and then compared with the results of phenotypic identification techniques. Two hundred and forty two yeasts were isolated and phenotypically identified as follows: Candida albicans (105), Candida tropicalis (62), Candida parapsilosis (28), Candida glabrata (19), Candida krusei (8), Candida guilliermondii (5) and Candida spp. (15). Samples from the three most frequent species isolated were selected randomly in order to compare the phenotypic and genotypic analyses. Genotypic analysis using RAPD primer M13 (F/R) displayed the best results of all test samples. There was both agreement and consistency between phenotypic and genotypic analysis using RAPD, demonstrating that is possible to apply this method for the identification of Candida species.

Candida↗

Mechanisms of host defense against Candida species. I. Phagocytosis by monocytes and monocyte-derived macrophages.

We studied the biochemical basis of phagocytosis of Candida albicans, a serious pathogen, and Candida parapsilosis, which is rarely pathogenic, by human monocytes (Mo) and monocyte-derived macrophages (MDM). Optimal phagocytosis of both species by Mo required the presence of extracellular Ca2+ and opsonization through both the classic and alternative complement pathways. Serum-opsonized Candida were ingested equally by Mo and MDM; unopsonized Candida were phagocytosed only by macrophages, and uptake began slowly. This opsonin-independent phagocytosis required Ca2+ and could be blocked by yeast mannan or mannose-BSA conjugate, suggesting a role for the mannose receptor. Opsonized Candida elicited a vigorous increase in the concentration of [Ca2+]i in Mo and MDM, but no Ca2+ transient was detected in MDM stimulated with unopsonized Candida. Pretreatment of MDM with ionomycin to increase [Ca2+]i had no effect on phagocytosis of unopsonized Candida. Addition of 5 mM EGTA completely inhibited changes in [Ca2+]i in Mo and MDM, suggesting that the Ca2+ transient induced by opsonized Candida is due to an influx of extracellular Ca2+. Differences in pathogenicity between the two Candida species could not be explained by differences in any aspect of phagocytosis. Uptake mediated by the macrophage mannose receptor could play a role in clearance of Candida under opsonin-poor conditions.

Calcium↗

Candida infections in bone marrow transplant recipients.

We studied the incidence, outcome and risk factors for systemic Candida infection in 665 recipients of allogeneic, syngeneic and autologous bone marrow transplantations (BMT) between 1979 and 1987. Systemic Candida infection, defined as occurrence of one or more positive blood or CSF cultures for Candida sp., or presence of Candida sp. in culture or biopsy of deep tissue, was detected in 76 patients (12.5%) in the first year following BMT. Candida infection was independently associated with increasing age (p less than 0.0001), detection of one or more positive surveillance cultures for Candida sp. (p less than 0.0001), increased duration of granulocytopenia (p = 0.0005) and total body irradiation as part of the preparative regimen compared with chemotherapy only or chemotherapy and total lymphoid irradiation (p = 0.02). Other patient characteristics including underlying disease, origin of graft, recipient sex, graft-versus-host disease (GVHD) prophylaxis and occurrence of acute GVHD or chronic GVHD were not independently associated with Candida infection following BMT: 60/76 patients with Candida infections have died, and in 19/60 cases death could be directly attributed to Candida infection. Awareness of the serious nature and the risk features for Candida infections may be useful in developing strategies of prevention and treatment.

Adolescent↗

Neutrophil-mediated protection of cultured human vascular endothelial cells from damage by growing Candida albicans hyphae.

Interactions were studied between human neutrophils and cultured human umbilical vein endothelial cells invaded by Candida albicans. In the absence of neutrophils, progressive Candida germination and hyphal growth extensively damaged endothelial cell monolayers over a period of 4 to 6 hours, as determined both by morphological changes and release of 51Cr from radiolabeled endothelial cells. Monolayers were completely destroyed and replaced by hyphae after 18 hours of incubation. In contrast, when added 2 hours after the monolayers had been infected with Candida, neutrophils selectively migrated toward and attached to hyphae at points of hyphal penetration into individual endothelial cells (observed by time-lapse video-microscopy). Attached neutrophils spread over hyphal surfaces both within and beneath the endothelial cells; neutrophil recruitment to initial sites of leukocyte-Candida-endothelial cell interactions continued throughout the first 60 minutes of observation. Neutrophil spreading and stasis were observed only along Candida hyphae and at sites of Candida-endothelial cell interactions. These events resulted in 58.0% killing of Candida at 2 hours and subsequent clearance of Candida from endothelial cell monolayers, as determined by microcolony counts and morphological observation. On introduction of additional neutrophils to yield higher ratios of neutrophils to endothelial cells (10 neutrophils:1 endothelial cell), neutrophil migration toward hyphal elements continued. Despite retraction or displacement of occasional endothelial cells by invading Candida and neutrophils, most endothelial cells remained intact, viable, and motile as verified both by morphological observations and measurement of 51Cr release from radiolabeled monolayers. From these studies, we conclude that neutrophils are capable of killing Candida hyphae selectively within human vascular endothelial cell monolayers and may have protective rather than detrimental effects on endothelial cell integrity.

Candida albicans↗

In vitro activity of clotrimazole for Candida strains isolated from recent patient samples.

Minimum inhibitory concentrations (MICs) of clotrimazole (CAS 23593-75-1, Bay 5097, clo) were determined for 142 clinical Candida isolates obtained between 1992 and 1994, including the species Candida albicans (96 strains), Candida glabrata (12 strains), Candida krusei (12 strains), and Candida tropicalis (12 strains). For some of the Candida isolates of all four species, the MICs of amphotericin B and fluconazole were also determined. No MICs of clo of > 4 micrograms/ml were found for all four Candida species, the median of MICs of clo for Candida albicans being 0.03 micrograms/ml with a range from < 0.015 to 4 micrograms/ml. The MICs of clo for Candida albicans were on average 2 log stages below the MICs of fluconazole and 1 log step below the MICs of amphotericin B on the microgram/ml level. Taking account of the present in vitro resistance situation, clo is a highly active antimycotic substance for isolates of all Candida species with human pathogenetic relevance.

Antifungal Agents↗

Candida sepsis during total parenteral nutrition: An endogenous infection indicating the severity of patients' disease state.

Between 1986 and 1989 we encountered 33 episodes of candida sepsis among 1169 patients receiving TPN for a total of 23350 days (2.8% candida infection rate). Total hospital stay averaged 78 (range 10-230) days and patients received TPN for an average of 21.5 (range 3-83) days before developing candida sepsis. Candida sepsis developed in 8 patients (26.6%) hospitalised in an ICU; 6 patients (20%) receiving high doses of glucocorticoids, 5 patients (16.6%) treated by cytotoxic agents; 23 patients (76.6%) received various combinations of broad-spectrum antibiotics. The number of tubes going in or out numbered an average of 3.6/patient (peripheral and/or central I.V.; endotracheal; tracheostomy; urinary catheter; arterial line; abdominal or chest drains). 18 patients underwent 38 (2.1/patient) operative procedures. 20 patients (66%) suffered fron mono- or polymicrobial bacterial sepsis in addition to candida sepsis, 16 of them metachronously. Candida species isolated were C. albicans - 14 patients; C. tropicalis - 6 patients; C. parapsylosis - 6 patients; not specified - 4 patients. In addition to positive blood cultures we found positive candida cultures in urine, peritoneal cavity, chest cavity, wounds, respiratory tract, intravascular catheters, often in more than one site per patient. All patients were treated with Amphotericin at an average dose of 770 mg/patient. Mortality rate in patients with candida sepsis was 33%. TPN associated candida sepsis seems to be an endogenous self-infecting process in a select group of severely injured-infected-depleted-immunosuppressed patients and is thus completely different from the usual exogenous bacterial TPN associated sepsis. The major risk factors for fungaemia and candida sepsis are the combination of severe underlying disease state, multiple surgical interventions and intravascular lines, the use of broad spectrum antibiotics, TPN, injury and malnutrition associated immunosuppression, multiple tubes and catheters, and intra-abdominal or intra-thoracic infection.

Journal Article↗