Comparison of Crystal Violet Staining Assay and XTT Methods in the Evaluation of Biofilm Formation in Candida parapsilosis Candidemia Isolates

O. Dogan, N. Atac, G. Babuccu, F. Can
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引用次数: 1

Abstract

Objective: Biofilm formation is one of the most important virulence factors of Candida species which leads to permanent infection foci by adhering to foreign materials and which are difficult to treat. Candida parapsilosis, which is one of the most common causes of candidemia in our country, is frequently isolated as a causative agent in catheterrelated infections. The most commonly used methods for evaluating the biofilm formation of Candida species are measuring cell viability with XTT (2,3-bis (2-methoxy-4-nitro-5sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide) and evaluating the total biofilm mass with crystal violet (CV). The aim of this study is to evaluate the biofilm formation ability of C. parapsilosis candidemia isolates by XTT and (CV) methods and compare these methods with each other. Materials and Methods: C. parapsilosis isolates sent from various hospitals between 2015 and 2019 were included in the study retrospectively, and the species-level identification was performed using the matrixassisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) The biofilm formation of the isolates was compared based on the optical density (OD) values obtained by crystal violet and XTT methods. The biofilm formation of the isolates was evaluated by categorizing them into low, medium and high biofilm groups as ± 20% according to the median value of all strains. Results: Totally, 79 C. parapsilosis candidemia isolates were included in this study and categorical compatibility between CV and XTT methods in low, medium and high biofilm groups was found as 69.6%, 60.6% and 73.9%, respectively. The OD values of the XTT method in the high biofilm group were found statistically significantly higher when compared with the values from the CV method. Conclusion: The compatibility of XTT and crystal violet methods in terms of biofilm measurement in C. parapsilosis isolates was considered acceptable, and no major variations were detected between the categories. According to these results, when evaluating the biofilm levels of C. parapsilosis isolates, high OD values obtained by the XTT method should be confirmed with the CV method.
结晶紫染色法与XTT法评价假丝酵母假丝菌分离株生物膜形成的比较
目的:生物膜形成是念珠菌最重要的毒力因素之一,念珠菌黏附异物形成永久性感染灶,治疗困难。假丝酵母菌伴假丝酵母菌病是我国最常见的念珠菌病之一,在导管相关感染中经常被分离出来作为病原体。评价念珠菌生物膜形成最常用的方法是用XTT(2,3-二(2-甲氧基-4-硝基-5-巯基)-5-[(苯胺)羰基]- 2h -氢氧化四氮唑)测定细胞活力和用结晶紫(CV)测定生物膜总质量。本研究的目的是通过XTT和(CV)两种方法评价假丝酵素假丝菌分离株的生物膜形成能力,并进行比较。材料与方法:回顾性收集2015 - 2019年各医院收治的假假梭菌分离株,采用基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF/MS)进行种级鉴定,并根据结晶紫法和XTT法获得的光密度(OD)值比较分离株的生物膜形成情况。根据所有菌株的中位数,将其分为低、中、高生物膜组(±20%),评价菌株的生物膜形成情况。结果:本研究共检获79株假丝酵菌,CV法和XTT法在低、中、高生物膜组的分类相容性分别为69.6%、60.6%和73.9%。高生物膜组XTT法OD值高于CV法OD值,差异有统计学意义。结论:XTT法和结晶紫法在假梭菌生物膜测定方面的相容性可接受,两种方法间无明显差异。综上所述,在评价副青枯病分离物的生物膜水平时,XTT法获得的较高OD值应与CV法一致。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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