Journal of Paramedical Sciences & Rehabilitation

Journal of Paramedical Sciences & Rehabilitation

Identification of Biofilm Formation Genes in Candida albicans and Evaluation of Iron Nanoparticles on the Expression of als gene by Real Time-PCR

Document Type : Original Article

Authors
L Hosseinzadeh
K Amini
Department of Microbiology, School of Basic Sciences, Saveh Branch, Islamic Azad University, Saveh, Iran.
10.22038/jpsr.2019.30557.1777
Abstract
Purpose:
Approximately three-fourths of women experience an episode of vaginal candidiasis. One of the virulence factors, is the ability of attachment of C. albicans to the host tissue through factors such as sticky molecules and proteins that accelerate this connection. The aim of this study was to investigate the frequency of biofilm formation genes in C. albicans strains isolated from vaginosis and to compare the expression of als gene in strains treated with iron nanoparticles in the in vitro using real time -PCR.
Methods:
In this descriptive cross-sectional study conducted in Shariati Hospital during a 6-months period, 100 isolates Candidafrom vaginal specimens were evaluated. After identification of C. albicans strains using standard biochemical and microbiological tests, genomic DNA extraction was performed using the CinnaGen Company Kit. Multiplex-PCR assays were performed to detect als, hwp1 and hyr genes. Also, to investigate the effect of iron nanoparticle on expression of als gene, RNA, all isolates containing this gene were extracted using Cinnapure kit. After cDNA synthesis, real time PCR test was performed.
Results:
Molecular analysis results showed that from the total of 60 isolates of C. albicans, 34 isolates (56.66%) had the hwp gene, 46 isolates (76.66%) had Als gene and 33 isolates (55%) had Hyr gene. The results of the asl melting curve in clinical strains of C. albicans and treated with iron nanoparticles showed that the main peak for this gene occurs at 85.34 ° C.
Conclusions:
Als expression in the treated isolates was lower than untreated, which indicates the positive role of the treatment in reducing Als gene expression in these isolates.
Keywords
Biofilms
Candida albicans
Vaginosis
Iron nanoparticles
Real Time-PCR
  1. Alli J, Okonko I, Odu N, Kolade A, et al. Detection and prevalence of Candida isolates among patients in Ibadan, Southwestern Nigeria. Journal of Microbiology and Biotechnology Research 2017; 1(3): 176-184.
  2. Gaydos CA, Beqaj S, Schwebke JR, Lebed J, TE, et al. Clinical Validation of a Test forthe Diagnosis of Vaginitis. Obstetrics and gynecology 2017; 130(1): 181-189.
  3. Jaeger M, Plantinga TS, Joosten LA, Kullberg B-J, et al. Genetic basis for recurrent vulvo-vaginal candidiasis. Current infectious disease reports 2013; 15(2): 136-142.
  4. Nakajima S, Harrison O, Linehan JL, Belkaid Y. Candida albicans colonization exacerbates skin inflammation in a murine model of psoriasis. Am Assoc Immnol; 2017.
  5. Calderone RA, Fonzi WA. Virulence factors of Candida albicans. Trends in microbiology 2001; 9(7): 327-335.
  6. Pakdel M, Zarrabi M, Asgarani E, Mohamadi P, et al. Identification of als1 and hwp1 Genes in Candida albicans Isolated Vaginal Infection. 2014; G3. [Persian]
  7. Richter SS, Galask RP, Messer SA, Hollis RJ, et al. Antifungal susceptibilities of Candida species causing vulvovaginitis and epidemiology of recurrent cases. Journal of Clinical Microbiology 2005; 43(5): 2155-62.
  8. Lynch M, Sobel J, Fidel Jr P. Role of antifungal drug resistance in the pathogenesis of recurrent vulvovaginal candidiasis. Journal of medical and veterinary mycology 1996; 34(5): 337-9. 
  9. Van de Ven H, Paulussen C, Feijens P, Matheeussen A, Rombaut P, Kayaert P, et al. PLGA nanoparticles and nanosuspensions with amphotericin B: Potent in vitro and in vivo alternatives to Fungizone and AmBisome. Journal of controlled release 2012; 161(3): 795-803.
  10. Qian J, Pan C, Liang C. Antimicrobial activity of Fe‐loaded chitosan nanoparticles. Engineering in Life Sciences. 2017. 
  11. Wangsanut T, Ghosh AK, Metzger PG, Fonzi WA, et al. Grf10 and Bas1 Regulate Transcription of Adenylate and One-Carbon Biosynthesis Genes and Affect Virulence in the Human Fungal Pathogen Candida albicans. mSphere 2017; 2(4): e00161-17. 
  12. Tsang PW, Bandara HM, Fong WP. Purpurin suppresses Candida albicans biofilm formation and hyphal development. PLoS One. 2012 Nov 30; 7(11): e50866 
  13. Hosseini SS, Ghaemi E, Noroozi A, Niknejad F. Zinc Oxide and ALS3 Gene Expression in Candida albicans Strains from Urinary Tract Infections. Nanoparticles Inhibition of Initial Adhesion and ALS1 . Mycopathologia. 2019 Apr;184(2):261-271.
  14. Mahmoudi Rad M, Zafarghandi AS, Amel Zabihi M, Tavallaee M, Mirdamadi Y. Identification of Candida species associated with vulvovaginal candidiasis by multiplex PCR. Infectious diseases in obstetrics and gynecology 2012: 872169. 
  15. Adesiji Y, Ndukwe N, Okanlawon B. Isolation and antifungal sensitivity to Candida isolates in young females. Open Medicine 2011; 6(2): 172-176.
  16. Ayatollahi Mousavi SA, Karami Robati A, Madani M, Hadizadeh S. Prevalence of Systemic fungal Infections in Kerman Teaching Hospitals. Journal of Mazandaran University of Medical Sciences 2013; 22(97): 105-112. 
  17. Hurley R, de Louvois J. Candida vaginitis. Postgraduate medical journal.1979; 55(647): 645.
  18. Trama JP, Adelson ME, Raphaelli I, Stemmer SM, et al. Detection of Candida species in vaginal samples in a clinical laboratory setting. Infectious diseases in obstetrics and gynecology 2005; 13(2): 63-7.
  19. Meisami F. A study common causes of bacterial vaginitis in woman referred to Mirza kochak khan hospital, Tehran, 2011. J Med Sci 1992; 3: 51-58.
  20. Holland J, Young M, Lee O, Chen SC. Vulvovaginal carriage of yeasts other than Candida albicans. Sexually transmitted infections 2003; 79(3): 249-50.
  21. Solanki A, Mathur D, Joshi K. Bacterial, fungal and parasitic flora in vaginitis. Journal of the Indian Medical Association 1983; 81(9-10): 151-3.
  22. Foladvand M, Naeimi B, Haghighi M. Identification of candida species isolated of vaginitis in woman referred to specialized hospital Bushehr. J Univ Med Sci & Health Services of Bushehr 2011; 2: 113-108. 
  23. Jamilian M, Mashad A, Sarmadi F, Ghaznavi-Rad A, et al, Ghena. Vulvovaginitis in non-pregnant woman 15-50 referred to Taleghani hospital Arak, 2005. 2006; 2: 1-7.
  24. Mahmoudabadi AZ, Najafyan M, Alidadi M. Clinical study of Candida vaginitis in Ahvaz, Iran and susceptibility of agents to topical antifungal. Pak J Med Sci 2010; 26(3): 607-610.
  25. Sobel JD, Brooker D, Stein GE, Thomason JL, et al. Single oral dose fluconazole compared with conventional clotrimazole topical therapy of Candida vaginitis. American journal of obstetrics and gynecology 1995; 172(4): 1263-1268.
  26. Rahimi H, Roudbar Ms, Kachouei R, Roudbari M. Expression of Candida albicans als 2 and als 9 genes isolated from women with vaginal candidiasis by RT-PCR. 2013.
  27. Inci M, Atalay MA, Özer B, Evirgen Ö, et al. Investigations of ALS1 and HWP1 genes in clinical isolates of Candida albicans. Turkish Journal of Medical Sciences 2013; 43(1): 125-130.
  28. Nailis H, Vandenbroucke R, Tilleman K, Deforce D, et al. Monitoring ALS1 and ALS3 gene expression during in vitro Candida albicans biofilm formation under continuous flow conditions. Mycopathologia 2009; 167(1): 9.
  29. Nas T, Kalkanci A, Fidan I, Hizel K, et al. Expression of ALS1, HWP1 and SAP4 genes in Candida albicans strains isolated from women with vaginitis. Folia Microbiol (Praha) 2008; 53(2): 179-183.
  30. Ghani S, Rafiee B, Sadeghi D, Ahsani M. Biosynthesis of Iron Nano-Particles by Bacillus Megaterium and Its Anti-Bacterial Properties. Journal of Babol University of Medical Sciences 2017; 19(7): 13-19.
Journal of Paramedical Sciences & Rehabilitation
Volume 8, Issue 3
Autumn 2019
Pages 25-33