Molecular and phenotypic identification of Candida isolates from pregnant women in Ogbomoso, Southwestern Nigeria
DOI:
https://doi.org/10.18203/2320-1770.ijrcog20160363Keywords:
Candida isolates, Pregnant women, Molecular identificationAbstract
Background: Vulvovaginal candidiasis (VVC), often referred to as a yeast infection is a common gynaecologic disease, affecting 3 out of 4 women in their lifetimes. More than 40% of affected women will have 2 or more VVC episodes, and infection occurs more frequently in pregnant women. This study was carried out to provide information on the appropriate diagnostic method required to differentiate the causative agents of VVC among pregnant women.
Methods: In this study, vaginal specimens were collected from one hundred (100) pregnant women aged between 17-44 years and of gestation age of 14-36 weeks who were attending antenatal clinic at LAUTECH Teaching Hospital, Ogbomoso. The species identification was performed using chromogenic medium, induction of fungal germ tube formation, and PCR using universal primers of internal transcriber spacer (ITS1 and ITS4); (ITS1 [5′-TCCGTAGGTGAACCTGCGG-3’] and ITS4 [5′-TCCTCCGCTTATTGATATGC-3’]) and Candida albicans-specific primers [5’-GGTTTGCTTGAAAGACGGTAG-3’] and [5’-AGTTTGAAGATATACGTGGTAG-3’] that target sequences site of the intergenic spacer region (ITS) of the fungal rRNA genes (18S and 28S) were used for this assay.
Results: Forty (40) Candida species from 100 specimens were isolated in Saboraud dextrose agar (SDA) medium. Of 19 strains of C. albicans that were identified by chromogenic agar (CHROMagar), 17 were confirmed as true positive by PCR while 2 were false positive. The CHROMagar had 89.4% sensitivity and 90.4% specificity. In comparison, GTT was better in correctly identifying those strains that were confirmed as C. albicans by PCR (Sensitivity=94%) while CHROMagar was better in identifying the strains that were not confirmed as C. albicans by PCR (Specificity=90.4%).
Conclusions: The combine uses of chromogenic agar and PCR have the advantage of efficient differentiation and identification of Candida species.References
Eloy O, Marque S, Batterel F, Stephan F, Costa JM, Laserre V, Bretagne S. Uniform distribution of three Candida albicans microsatellite markers in two French ICU populations supports a lack of nosocomial cross-contamination. BMC Infect Dis. 2006;6:162-3.
Kamiya A, Tomita Y, Kikuchi A, Knabe T. Epidemiological study of Candida species in cutaneous candidiasis based on PCR using a primer mix specific for the DNA topoisomerase II gene. J Dermatol Sci. 2005;21-8.
Anaissie EJ, McGinnis MR, Pfaller MA. Clinical Mycology, Philadelphia: Elsevier Sciences; 2003.
Rippon JW. Medical Mycology. The pathogenic fungi & actinomycetes. Philadelphia:W.B. Saunders Co; 1988.
Edmond MB, Wallace SE, Mc Clish DK, Pfaller MA, Jones RN, Wenzel RP. Nosocomial bloodstream infections in United States hospitals: a three-year analysis. Clin Infec Dis. 1999;29(2):239-44.
Hedayati T, Shafiei G. Candidiasis. eMedicine Specialties. 2010.
Das Neves J, Pinto E, Teixeira B, Dias G, Rocha P, Cunha T. Local treatment of vulvovaginal candidosis: general and practical considerations. Drugs. 2008;68(13):1787-802.
Eschenbach DA. Chronic vulvovaginal candidiasis. N Engl J Med. 2004;351(9):851-2.
Monif GR, Baker DA. Candida albicans. In: Monif GR, Baker DA, editors. Infectious diseases in obstetrics and gynecology. 5th ed. New York, NY: Parthenon Press; 2003.
Baron EJ, Cassell GH, Duffy LB, Eschenbach JR, Greenwood SM, Harvey NE. Laboratory diagnosis of female genital tract infections. In: Baron EJ, editor. Cumulative techniques and procedures in clinical microbiology (Cumitech); 1993.
Carlson P, Richardson M, Paavonen J. Evaluation of the oricult-N Dipslide for laboratory diagnosis of vaginal candidiasis. J Clin Microbiol. 2000;38(3):1063-5.
Guthrie BL, Kiariae JN, Morrison S, John-Stewart GC, Kimuthia J, Whittington WLH, Farquhar C. Sexually transmitted infections among HIV-1-discordant couples. PLoS One. 2009;4(12):8276.
Ahmad S, Khan Z, Mustafa AS, Khan ZU. Seminested PCR for diagnosis of candidemia: comparison with culture, antigen detection, and biochemical methods for species identification. J Clin Microbiol. 2002;40(7):2483-9.
Cheesbrough M. Microbiological test in; District laboratory practice for tropical countries part 2, Cambridge Low Price Edition, Cambridge University Press; 2000.
Baker F. Handbook of bacteriological techniques. 2nd ed. Butterworth Co. ltd London; 1967.
Mohamed SA, Al-Ahmadey ZZ. Biofilm formation and antifungal susceptibility of Candida isolates from various clinical specimens. Brit Micribiol Res J. 2013;3(4):590-601.
Metwally L, Fairly DJ, Coyle PV,Hay RJ, Hedderwick S, McCloskey B,O’Neill HJ,Webb CH,Elbaz Wand McMullan R. Improving molecular detection of Candida DNA in whole blood: comparison of seven fungal DNA extraction protocols using real-time PCR. J. Med. Microbiol. 2008;57:296-303.
Mousavi SA, Salari S, Rezaie S, Shahabi N, Nejad NS, Hadizadeh S, Kamyabi H, Aghasi H. Identification of Candida species isolated from oral colonization in Iranian HIV-positive patients, by PCR-RFLP method. Jundishapur J Microbiol. 2012;5(1):336-40.
Akinbami AN, Babalola GO, Shittu MO, Tijani AM, Adekola SA. Detection and epidemiology of vulvovaginal candidiasis among asymptomatic pregnant women attending a tertiary hospital in Ogbomoso, Nigeria. Intern J Biomed Res. 2015;6(07):518-23.
Merlino J, Tambosis E, Veal D. Chromogenic tube test for presumptive identification or confirmation of isolates as Candida albicans. J Clin Microbiol. 1998;36(4):1157-9.
Kazemi A, Falahati M, Hajipoor A, Jafari A, Asghar-zadeh M. Comparison of phenotypic tests and PCR to detect Candida albicans from vaginal specimens. Jundishapur J Microbiol. 2013;6(2):122-6.
Yucesoy M, Esen N, Yulug N. Use of chromogenic tube and methyl blue-sabouraud agar for the identification of Candida albicans strains. Kobe J Med Sci. 2001;47(4):161-7.
Mahens B, Stehr F, Schafer W, Neuber K. Comparison of standard phenotypic assays with a PCR method to discriminate Candida albicans and C. dubliniensis. Mycoses. 2005;48(1):55-61.
Ainscough S, Kibbler CC. An evaluation of the cost-effectiveness of using CHROM agar for yeast identification in a routine microbiology laboratory. J Med Microbiol. 1998;47(7):623-8.
Hoppe JE, Frey P. Evaluation of six commercial tests and the germ-tube test for presumptive identification of Candida albicans. Eur J Clin Microbiol Infect Dis. 1999;18(3):188-91.
