Evaluation of Peptide Nucleic Acid Fluorescent In Situ Hybridization (PNA FISH) Method in the Identification of Candida Species Isolated From Blood Cultures

In recent years, increased number of patients who are hospitalized in intensive care units, received immunosuppressive therapy and treated with broad-spectrum antibiotics that can lead an increase in the incidence of systemic candidiasis. In these patients, the most common clinical manifestation is candidemia. Since the identification of Candida species isolated from blood cultures is time consuming by conventional (morphological and biochemical) methods, rapid, reliable and accurate methods are needed. For this purpose novel systems have been developed to identify the agent directly. The aim of this study was to evaluate the peptide nucleic acid fluorescent in situ hybridization (PNA FISH) method for the identification of Candida species by comparing with the conventional methods. A total of 50 patients who were admitted to Erciyes University Medical Faculty Hospital clinics and followed with prediagnosis of systemic fungal infections whose blood cultures were positive for the yeasts between July 2011 and July 2012 were included in the study. The conventional identification of Candida isolates was performed by considering macroscopic and microscopic morphology, germ tube test, cycloheximide sensitivity, urease activity and carbohydrate assimilation patterns with API 20C AUX (bioMerieux, France) test. PNA FISH method was conducted by the use of a commercial kit namely Yeast Traffic Light PNA FISH (AdvanDx, USA). According to morphological and biochemical characteristics (conventional methods), 19 (38%) out of 50 Candida isolates were identified as C.albicans, 12 (24%) as C.glabrata, five (10%) as C.parapsilosis, five (10%) as C.kefyr, four (8%) as C.krusei, two (4%) as C.guilliermondii, two (4%) as C.tropicalis and one (2%) as C.lusitaniae. On the other hand, 24 (48%) of the isolates were identified as C.albicans/C.parapsilosis (with green fluorescence), 16 (32%) as C.glabrata/C.krusei (with red fluorescence) and one (%2) as C.tropicalis (with yellow fluorescence) properly, however one C.tropicalis strain was misidentified as C.albicans by PNA FISH method. Other eight (16%) strains which were not presented in the evaluation panel of PNA FISH kit (5 C.kefyr, 2 C.guillermondii and 1 C.lusitaniae), gave no fluorescence and determined as other Candida spp. According to this, when the species that could be detected with the kit (C.albicans, C.parapsilosis, C.glabrata, C.krusei and C.tropicalis) were considered, the concordance rate with the conventional methods was determined as 97.6% (41/42) and the total evaluation rate for all the species was 84% (41/50). In conclusion, the most frequent isolated species from blood cultures in our hospital was C.albicans, followed by C.glabrata and C.parapsilosis. Since PNA FISH testing is a practical, reliable and rapid (resulted in 90 minutes) method for the identification of Candida strains at species level isolated from blood cultures, it was thought to be useful in routine laboratories. However, further comparative studies are required with large number of strains with the consideration of cost-effectiveness

Late neurologic symptoms of carbonmonoxide intoxication: A case report

Karbonmonoksit; akut ya da kronik zehirlenmeye neden olabilen renksiz, kokusuz bir gazdır. Karbon monoksit zehirlenmesine bağlı beklenmeyen ölümler özellikle kış aylarında artmaktadır. Karbon monoksit dokulara oksijen sunumunu azaltır. Zehirlenmeden günler veya haftalar sonra görülen geç nörolojik bulgular karbonmonoksit zehirlenmesinin en önemli komplikasyonlarıdır. On üç yaşında kız çocuğu ajitasyon, gözlerde sola kayma, kol ve bacaklarında istemsiz hareketler, idrar kaçırma şikâyetleri ile çocuk acil polikliniğine başvurdu. Öyküsünden, beş gün önce şofbenden zehirlendiği öğrenildi. Yapılan ayrıntılı değerlendirmeler sonucu karbon monoksit zehirlenmesinin geç komplikasyonu olarak değerlendirilen hastanın bulguları uygun destek tedavisinden sonra düzeldi.Carbon monoxide is colourless, odourless gas that can cause acute or chronic intoxication. Carbonmonoxide reduces oxygen delivery of tissues. Late neurologic symptoms of carbonmonoxide intoxication can occur days or even weeks after the patient has made a seemingly good recovery from the acute poisoning. The patient, a 13-years old female girl admitted to hospital with disorientation, hemiparesis, choreoathetosis. From her history, we learned that she was poisoned from geyser five days ago. On detailed evaluation the patient is diagnosed as the late neurological signs of carbonmonoxide intoxication. Symptoms of patient is improved after appropriate supportive management

Mycoplasma pneumoniae'ya bagli yogun parapnömonik efüzyonlu bir olgu sunumu

Mycoplasma pneumoniae infection is usually mild course and self-limited. Parapneumonic effusion is not a common feature of M. pneumoniae. The treatment of parapneumonic effusion is macrolides and chest tube with adequate pleural drainage. We report here on a child with M. pneumoniae infection complicated by massive parapneumonic effusion. Protracted course of fever and respiratory distress was noted in patient. Pneumothorax was occured subsequent to chest tube drainage

Fungemia and Septic Arthritis Caused by Saprochaete capitata in a Patient with Fanconi Aplastic Anemia: A Case Report

Saprochaete capitata (formerly known as Blastoschizomyces capitatus, Trichosporon capitatum, Geotrichum capitatum) is a rare but emerging yeast-like fungus. It is commonly found in environmental sources and can be isolated from skin, gastrointestinal system and respiratory tract of healthy individuals as well. It mainly infects patients with hematological malignancies such as acute myeloid leukemia (AML), especially in the presence of neutropenia; and mortality rates are high in those patients. Although the data about the in vitro antifungal susceptibility are limited, it is being reported that amphotericin B and voriconazole are more effective on S.capitata isolates whereas caspofungin had no activity. Here, we report a case of fungemia and septic arthritis due to S.capitata in a patient with Fanconi aplastic anemia. A 22-year-old male patient with Fanconi aplastic anemia was hospitalized in our hematology department for bone marrow transplantation. Two days after the hospitalization, neutropenic fever developed and multiple nodules similar to candidiasis were detected in his liver with the whole abdomen magnetic resonance imaging (MRI). Caspofungin treatment (single 70 mg/kg loading dose, followed by 1 x 50 mg/kg/day) was started. The patient remained febrile, and his blood culture yielded S.capitata. The treatment regimen was changed to a combination of liposomal amphotericin B (3 mg/kg/day) and voriconazole (2 x 4 mg/kg/day). A few days later, pain and swelling came out on patient's left knee and he underwent a surgical process with the prediagnosis of septic arthritis. Culture of synovial fluid was also positive for S.capitata. On the 26th day of the hospitalization, the patient died due to sepsis and multiple organ failure. Patient's blood and synovial fluid samples were incubated in BacT/Alert automated blood culture system (bioMerieux, France). After receiving the growth signal, yeast cells were seen in Gram staining and cream-coloured, wrinkled, yeast-like colonies that were able to grow at 45 degrees C and resistant to cycloheximide were detected on Sabouraud dextrose agar (SDA). Urease test was negative, and according to API 20C AUX (bioMerieux, France) system, none of the carbonhydrates were utilized except glucose. The isolates that were able to produce annelloconidia in corn meal-Tween 80 agar slide culture were identified as S.capitata. The identification was further confirmed by DNA sequence analysis. Minimal inhibitory concentrations (MICs) of amphotericin B, fluconazole, voriconazole, and caspofungin were found to be 0.5 mu g/ml, 1.5 mu g/ml, 0.032 mu g/ml, and > 16 mu g/ml respectively. Repetitive sequence based PCR (rep-PCR) (DiversiLab system, bioMerieux, France) was used to determine clonal relatedness of the isolates from blood and synovial fluid samples. The isolates were indistinguishable (similarity coefficient > 97%) according to rep-PCR. In conclusion, S. capitata infections should be taken into consideration in the presence of fungemia and septic arthritis in hematological patients who receive caspofungin therapy