Background:Candida infection is a major cause of morbidity and mortality in immunocompromised patients; an accurate and early identification is a prerequisite need to be taken as an effective measure for the management of patients. The purpose of this study was to compare the conventional identification of Candida species with identification by Vitek-2 system and the antifungal susceptibility testing (AST) by broth microdilution method with Vitek-2 AST system.Materials and Methods:A total of 172 Candida isolates were subjected for identification by the conventional methods, Vitek-2 system, restriction fragment length polymorphism, and random amplified polymorphic DNA analysis. AST was carried out as per the Clinical and Laboratory Standards Institute M27-A3 document and by Vitek-2 system.Results:Candida albicans (82.51%) was the most common Candida species followed by Candida tropicalis (6.29%), Candida krusei (4.89%), Candida parapsilosis (3.49%), and Candida glabrata (2.79%). With Vitek-2 system, of the 172 isolates, 155 Candida isolates were correctly identified, 13 were misidentified, and four were with low discrimination. Whereas with conventional methods, 171 Candida isolates were correctly identified and only a single isolate of C. albicans was misidentified as C. tropicalis. The average measurement of agreement between the Vitek-2 system and conventional methods was >94%. Most of the isolates were susceptible to fluconazole (88.95%) and amphotericin B (97.67%). The measurement of agreement between the methods of AST was >94% for fluconazole and >99% for amphotericin B, which was statistically significant (P < 0.01).Conclusion:The study confirmed the importance and reliability of conventional and molecular methods, and the acceptable agreements suggest Vitek-2 system an alternative method for speciation and sensitivity testing of Candida species infections.
HIV related opportunistic fungal infections (OFIs) continue to cause morbidity and mortality in HIV infected patients. The objective for this prospective study is to elucidate the prevalence and spectrum of common OFIs in HIV/AIDS patients in north India. Relevant clinical samples were collected from symptomatic HIV positive patients (n = 280) of all age groups and both sexes and subjected to direct microscopy and fungal culture. Identification as well as speciation of the fungal isolates was done as per the standard recommended methods. CD4+T cell counts were determined by flow cytometry using Fluorescent Activated Cell Sorter Count system. 215 fungal isolates were isolated with the isolation rate of 41.1%. Candida species (86.5%) were the commonest followed by Aspergillus (6.5%), Cryptococcus (3.3%), Penicillium (1.9%), and Alternaria and Rhodotorula spp. (0.9% each). Among Candida species, Candida albicans (75.8%) was the most prevalent species followed by C. tropicalis (9.7%), C. krusei (6.4%), C. glabrata (4.3%), C. parapsilosis (2.7%), and C. kefyr (1.1%). Study demonstrates that the oropharyngeal candidiasis is the commonest among different OFIs and would help to increase the awareness of clinicians in diagnosis and early treatment of these infections helping in the proper management of the patients especially in resource limited countries like ours.
The purpose of this prospective study was to isolate, speciate, and determine antifungal susceptibility and virulence patterns of Candida species recovered from the intensive care units (ICUs) in an Indian hospital. Study included 125 medical/postoperative patients admitted to ICU. Identification and speciation of yeast isolates were done by the biochemical methods. Antifungal susceptibility was done by broth microdilution method. Virulence testing of Candida species was done by phospholipase, proteinase, and adherence assay. A total of 103 Candida isolates were isolated; C. tropicalis was the predominant species (40.7%), followed by C. albicans (38.83 %), C. glabrata (11.65%), C. parapsilosis (3.88%), and 1.94% each of C. krusei, C. kefyr, and C. sphaerica. 60 Candida isolates (58.25%) showed resistance to fluconazole, while 7 (6.7%) isolates showed resistance to amphotericin B. Phospholipase and proteinase activities were seen in 73.8% and 55.3% Candida isolates with different species showing a wide range of activities, while 68.9% Candida isolates showed {4+} adherence activity. The present study revealed that nonalbicans Candida species (NAC spp.) caused most of the cases of Candidemia in the ICU patients. The isolation of C. tropicalis from a large number of cases highlights the ability of this pathogen to cause bloodstream infections. The presence of azole resistance is a matter of concern.
HIV has become a major health problem in India, patients commonly succumb to opportunistic infections (OIs), respiratory infections being an important cause of morbidity and their accurate diagnosis is still a challenge. Our aim was to study the occurrence of Pneumocystis pneumonia (PCP) in HIV/AIDS patients with respiratory complaints attending ART clinic and to compare various diagnostic methodologies. One hundred and twenty five HIV/AIDS patients presenting with respiratory symptoms like cough, fever, breathlessness etc, were enrolled, and induced sputum samples were collected. Samples were homogenized using glass beads and Dithiothretol. Smears were prepared and examined by Immunoflourescent staining (IFAT), Gomori methanamine silver staining (GMSS), Toludine blue O staining (TBO) and Giemsa staining for Pneumocystis jiroveci. Among the 125 patients who presented with respiratory complaints, 34 cases (27.2%) were diagnosed as having PCP. All 34 cases were detected by IFAT followed by GMSS, Giemsa and Toludine blue O staining in decreasing order. The mean CD4 count was 67.27cells/μl. PCP has become an important health problem in HIV/AIDS patients with low CD4 counts in India. IFAT remains the most sensitive method for the detection of this uncultivable organism. In resource poor settings where an immunoflourecent microscope is not available, diagnosis of PCP still remains problematic.
Background: Candidemia causing increased mortality rates and emergence of antifungal drug resistance needs an urgent intervention to salvage immunocompromised and severely ill patients. This study aimed to isolate and identify Candida species and evaluate their antifungal susceptibility profile from blood stream infections in children.Methods: Fungal cultures from blood recovered positive for yeasts were subcultured on Sabouraud dextrose agar. Suspected purified colonies of Candida were confirmed and identified upto species level by both conventional and automated techniques. Antifungal susceptibility testing of isolates was evaluated using agar based E-test method for fluconazole, voriconazole and caspofungin on Mueller-Hinton agar supplemented with 2% glucose.Results: Total of 43 isolates of Candida species were recovered from blood samples. Non albicans Candida species accounted for 88.30% of cases; whereas 11.60% of cases were caused by C. albicans, C. tropicalis (39%) was the most frequent isolate recovered in candidemia patients followed by C. parapsilosis (18%), C. albicans (12%), C. glabrata (12%), C. kefyr (9%), C. pelliculosa (5%), and C. krusei (5%). Antifungal susceptibility results revealed Caspofungin demonstrated good activity against all Candida spp. C. parapsilosis followed by C. tropicalis and C. glabrata demonstrated high resistance to fluconazole. For voriconazole, maximum resistance was shown by C. tropicalis as compared to others.Conclusions: Candidemia is a threatening prognostic sign in children and an important entity in our hospital. Identification of Candidaspecies and antifungal sensitivity testing is a must to select a suitable and effective antifungal therapy to abrogate the emerging resistance to antifungals.
The study was conducted to study the occurrence and clinical presentation of allergic fungal rhinosinusitis (AFRS), characterize the same, and correlate with the microbiological profile. Clinically suspected cases of fungal rhinosinusitis (FRS) depending upon their clinical presentation, nasal endoscopy, and radiological evidences were included. Relevant clinical samples were collected and subjected to direct microscopy and culture and histopathological examination. 35 patients were diagnosed to have AFRS. The average age was 28.4 years with a range of 18–48 years. Allergic mucin was seen in all the AFRS patients but fungal hyphae were detected in only 20%. 80% of cases were positive for IgE. All the patients had nasal obstruction followed by nasal discharge (62.8%). Polyps were seen in 95% (unilateral (48.57%) and bilateral (45.71%)), deviated nasal septum was seen in 28.57%, and greenish yellow secretion was seen in 17.14%. Direct microscopy and septate hyphae were positive in 71.42% of cases. 91.4% of cases were positive by culture. 5.7% yielded mixed growth of A. flavus and A. niger. Prompt clinical suspicion with specific signs and symptoms along with timely sampling of the adequate patient specimens and the optimal and timely processing by microscopy and culture and histopathological examination is a must for early diagnosis and management.
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