Premamalini Thayanidhi, Anitha Subramanian, Mohanapriya Kanniappan, Kindo Anupma Jyoti
Department of Microbiology, Sri Ramachandra Institute of Higher Education and Research, Chennai, Tamil Nadu, India.
J Lab Physicians. 2018 Oct-Dec;10(4):380-386. doi: 10.4103/JLP.JLP_37_18.
Invasive infections due to spp. have increased recently and are frequently associated with indwelling medical devices. Such infections which are associated with biofilm formation do not respond to the routinely used antifungal agents and are often persistent, associated with high mortality rate. Various methods have been described by researchers to evaluate and quantify the biofilm formation.
This study was conducted to compare two methods of biofilm production by sp, i.e., test tube method with crystal violet (CV) staining and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay.
Seventy-two clinical isolates of spp. collected from various sources were considered for the study. The identity of all the isolates was genotypically confirmed by -specific polymerase chain reaction (PCR). The isolates were further speciated phenotypically using biochemical profile and growth characteristics which identified the isolates as (64/72), (5/72), (2/72), and (1/72). Biofilm production was then evaluated and compared by test tube-CV method and MTT assay.
All the isolates produced biofilm by MTT assay, whereas only 42 (53.6%) of the isolates were detected to be biofilm producers by CV method. Furthermore, MTT assay could differentiate better between weak and moderate biofilm producers as compared to CV method.
Hence, MTT assay is a reliable method for quantification of biofilm produced by spp. using 96-well microtiter plate.
近期，由[具体菌种名称]引起的侵袭性感染有所增加，且常与留置医疗设备相关。此类与生物膜形成相关的感染对常规使用的抗真菌药物无反应，且往往持续存在，死亡率高。研究人员已描述了多种评估和量化生物膜形成的方法。
本研究旨在比较[具体菌种名称]产生生物膜的两种方法，即采用结晶紫（CV）染色的试管法和3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法。
本研究纳入了从不同来源收集的72株[具体菌种名称]临床分离株。所有分离株的身份通过[具体菌种名称]特异性聚合酶链反应（PCR）进行基因确认。利用生化特征和生长特性对分离株进行进一步的表型鉴定，确定分离株为[具体菌种名称1]（64/72）、[具体菌种名称2]（5/72）、[具体菌种名称3]（2/72）和[具体菌种名称4]（1/72）。然后通过试管-CV法和MTT法评估并比较生物膜的产生情况。
所有[具体菌种名称]分离株通过MTT法均产生生物膜，而通过CV法仅检测到42株（53.6%）分离株为生物膜产生菌。此外，与CV法相比，MTT法在区分弱生物膜产生菌和中等生物膜产生菌方面表现更佳。
因此，MTT法是一种使用96孔微量滴定板定量[具体菌种名称]产生生物膜的可靠方法。
