SDS-Page and numerical analysis of Candida albicans from human oral cavity and other anatomical sites

Abstract

The aim of the present research was to evaluate the protein polymorphism degrees among forty-eight C. albicans isolates from fourteen anatomical sites of clinical patients by polyacrylamide gel electrophoresis (SDS-PAGE) and numerical analyzes, in order to identify subspecies and their similarities in some infectious niches. Cell cultures were grown in YEPD medium, collected by centrifugation, and washed in cold saline solution. The whole-cell proteins were extracted by cell disruption using glass beads and submitted to SDS-PAGE technique. After electrophoresis, the protein bands were stained with coomassie-blue and analyzed by statistics package NTSYS-pc version 1.70 software. Similarity matrixes and dendrograms were generated by application of similarity coefficient of simple matching and UPGMA algorithm, respectively. The results obtained showed several C. albicans subtypes and their similarity degrees (80% to 100%). Such data showed that same patients may be infected by two or more C. albicans subtypes in certain anatomical sites (i.e. only in oral cavity of immunocompromised patients, blood, or tracheal secretion), or yet, two or more patients can be infected in identical anatomical sites (i.e. bronchial washing, urine, oral cavity, tracheal secretion, vaginal secretion, and healthy saliva) with a same C. albicans subtype. However, two or more patients also can show infections in corresponding sites (i.e. oral cavity of immunocompromised patients, blood, oropharyngeal secretion, oral cavity, tracheal secretion, vaginal secretion, and healthy saliva) by different C. albicans subtypes. Besides, two or more patients also can be infected with identical or different C. albicans subtypes in different anatomical sites (i.e.1. identical subtypes in vaginal secretion, tracheal secretion, and urine; abdominal secretion and spittle; drainage and oral cavity; catheter and healthy saliva -i.e.2. subtypes different in bronchial washing, oropharyngeal secretion, pulmonary secretion, oral cavity of immunocompromised patients, and blood). Complementary studies involving C. albicans sample isolated from several anatomical sites of immunocompetent or immunocompromised patients (before, during and after specifics therapies) and their families or hospital workers must be done in order to establish the sources of C. albicans colonization. The whole-cell proteins profile performed by SDS-PAGE associated with computerassisted numerical analysis may provide preliminary criteria for taxonomic and epidemiological studies of such microorganisms.

Total Protein Profile and Drug Resistance inCandida albicansIsolated from Clinical Samples

“…The efficacy of use of SDS-PAGE has also been described in aspergillosis in bronchogenic carcinoma [ 7 ]. Rodrigues et al [ 8 ] reported the use of SDS-PAGE for typing of C. albicans isolates. They found this method to be important for characterizing the yeast for epidemiological and taxonomic purposes.…”

“…The SDS-PAGE has been reported for epidemiological typing of Klebsiella spp. [ 6 ], aspergillosis [ 7 ], and C. albicans [ 8 ]. Ying et al [ 9 ] indicated that the fluconazole resistance of C. albicans has overexpression of ERG11 gene.…”

Antimicrobial effect of para-alkoxyphenylcarbamic acid esters containing substituted N-phenylpiperazine moiety

“…However, in hosts predisposed to candidiasis, for example very low birth weight, immunosupressive, antibiotic, and cytotoxic therapies, diabetes mellitus , organ transplant, tumors, immunocompromised patients affected by AIDS and others, these yeasts may act as pathogens. Commensal Candida species normally inhabit in the oral, nasal and aural cavity, vaginal canal, rectum, and gastrointestinal tract of host may begin the infectious process ( Diaz-Guerra et al , 1997 ; Hellstein et al , 1993 ; Miyasaka et al , 2008 ; Oliveira et al , 2011 ; Rodrigues et al , 2004 ).…”

Molecular fingerprinting methods for the discrimination between C. albicans and C. dubliniensis

“…While various studies have recommended the SDS‐PAGE as a useful technique for discriminating different Candida species, particularly C. albicans (Rosa et al , 2000; Boriollo et al , 2003; Rodrigues et al , 2004), little information is available on pertinent literature concerning the use of this technique for identification of C. dubliniensis . Multilocus enzyme electrophoresis (MLEE) and SDS‐PAGE were recently used to separate C. albicans and C. dubliniensis into different taxa (Rosa et al , 2003).…”

“…The sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) has emerged as one of the most versatile and inexpensive techniques for the separation and resolution of bacterial proteins that may be used for the establishment of the phenotypic variability and relationship within species (Kersters, 1985). Many studies have been used this technique for the identification of important strains (Cunningham and Noble, 1989; Rosa et al , 2000; Boriollo et al , 2003; Rosa et al , 2003; Rodrigues et al , 2004). The comparison of electrophoretic protein patterns has satisfactory taxonomic resolution, which may be applicable to the level of species, subspecies and biotypes (Kersters, 1985).…”