Fungal profile of Candidemia by multiplex polymerase chain reaction in a Tertiary Care Hospital
Rakesh Singh, S. Radha
E-mail – drrakesh1976@yahoo.com
Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education and Research 


Introduction: Candida albicans is the most common causative agent of candidemia world-wide. The fungal profile of candidemia varies geographically and is different from one hospital to another within the same region. Candida tropicalis is the most common etiological agent of candidemia in India although some studies reported C. albicans. Most of the laboratories are identifying the yeast by phenotypic techniques which is labour intensive and less specific as compared to polymerase chain reaction (PCR). Identification of the yeast by PCR using internal transcribed spacer (ITS) regions is highly specific. There is a limited literature available regarding the fungal profile of candidemia using PCR especially in the South India. 
Aim and Objective: To determine the fungal profile of candidemia cases by multiplex polymerase chain reaction in a tertiary care hospital

Methods: The study was conducted after getting Institute ethical clearance. Forty three yeast positive blood culture bottles were observed from 1st March 2016 to 31st January 2017 at the Department of Microbiology, Jawaharlal Institute of Postgraduate Medical Education and Research, Puducherry, India and included in the study. Species identification was made by multiplex PCR using ITS 1 and ITS 2 region of 18S, 5.8S and 28S nuclear rRNA gene with C. albicans, C. tropicalis, C. glabrata, C. parapsilosis complex, C. krusei primers as described by Carvalho et al. with minor modification.  Antifungal susceptibility was performed by disc diffusion method for fluconazole (25µg) and voriconazole (1µg) as per Clinical and Laboratory Standards Institute (CLSI) M44A2 guidelines. The categorical variables were expressed as frequencies & percentages.  

Results: Candidemia cases were seen in patients with age from 1 month to 78 years and with the median of 50 years. 37.2% of the isolates were from females and 62.8% of strains were from males. 23.3% of the isolates were obtained from patients between 51-60 years of age. 53.5% of the candidemia cases were admitted in ICU. The most common isolate was Candida tropicalis (34.9%) followed by Candida parapsilosis complex (30.2%), Candida albicans (16.3%), Candida glabrata (14.0%), and Candida krusei (4.7%). 23.3% strains were resistance and 14% were susceptible dose dependent to fluconazole whereas only 2.3% strains were resistance to voriconazole.  A single strain of Candida parapsilosis complex was observed to be resistant to both fluconazole and voriconazole. 
Conclusions: Candida tropicalis is the most common causative agent of candidemia in our tertiary care hospital. 23.3% of candidemia strains were resistance to fluconazole. 
References: 
1.	Tan BH, Chakrabarti A, Li RY, Patel AK, Watcharananan SP, Liu Z, et al. Incidence and species distribution of candidaemia in Asia: a laboratory-based surveillance study. Clin Microbiol Infect. 2015 Oct;21(10):946–53.
2.	CLSI. Methods for antifungal disc diffusion susceptibility testing of yeasts; Approved Guideline-Second edition. CLSI document M44-A2. Wayne, PA: Clinical and Laboratory Standards Institute; 2009
3.	Chander J, Singla N, Sidhu SK, Gombar S. Epidemiology of Candida blood stream infections: experience of a tertiary care centre in North India. J Infect Dev Ctries. 2013 Sep 16;7(9):670–5. 
4.	Banerjee B, R M SD, Baliga S. Clinico-microbiological study of candidemia in a tertiary care hospital of southern part of India. Iran J Microbiol. 2015 Feb;7(1):55–61. 
5.	Bhattacharjee P. Epidemiology and antifungal susceptibility of Candida species in a tertiary care hospital, Kolkata, India. Curr Med Mycol. 2016 Jun;2(2):20–7. 
6.	Singh R, Parija SC. Candida parapsilosis : an emerging fungal pathogen. Indian J Med Res. 2012 Oct 1;136(4):671.