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[作为医院病原体分离出的念珠菌菌株的表型和基因型鉴定]

[Phenotypic and genotypic identification of Candida strains isolated as nosocomial pathogens].

作者信息

Sahiner Fatih, Ergünay Koray, Ozyurt Mustafa, Ardıç Nurittin, Hoşbul Tuğrul, Haznedaroğlu Tunçer

机构信息

Gülhane Millitary Medical Academy, Haydarpaşa Training Hospital, Department of Medical Microbiology, İstanbul, Turkey.

出版信息

Mikrobiyol Bul. 2011 Jul;45(3):478-88.

Abstract

Over the last decade, there have been important changes in the epidemiology of Candida infections and antifungal agents used to treat these infections. In recent years, Candida species have emerged as important causes of invasive infections among patients in intensive care units. One of the main goals of this study was to evaluate the molecular epidemiology of infectious Candida species isolated in our hospital and accordingly supply data for hospital infection (HI) control. The other aim of this study was to evaluate effectiveness and practical applicability of traditional and molecular methods used to identify Candida isolates to the species level. A total of 77 Candida strains that were isolated from various clinical specimens of 60 hospitalized patients (29 male, 24 female; 7 were children) were included in the study. Fifty-seven (74%) of those isolates were defined as HI agents according to Centers for Disease Control and Prevention (CDC) criteria. The most common Candida species identified as agents of HI were C.albicans (22; 38.6%), followed by C.tropicalis (14; 24.6%), C.parapsilosis (13; 22.8%), C.glabrata (7; 12.3%) and Candida spp. (1; 1.75%). It was determined that bloodstream (26; 45.6%) and urinary tract infections (24; 42.1%) were the most frequently encountered nosocomial infections caused by Candida species. In addition it was detected that the most frequent causative agent of bloodstream infections was C.parapsilosis (10; 38.5%) and of urinary tract infections was C.albicans (12; 50%). The evaluation of advantages and disadvantages of traditional phenotypic methods [germ tube formation, chlamydospore formation in corn meal agar, growth at 45°C, colony characteristics on CHROMagar Candida medium, carbohydrate assimilation properties detected by API ID 32C (BioMerieux, France) system] and some molecular techniques [polymerase chain reaction (PCR) by using ITS-1, ITS-3 and ITS 4 primers, PCR-Restriction fragment length polymorphism (RFLP), PCRRFLP in which ITS1-ITS4 products cut by Msp I ve Bln I restriction enzymes] for the identification of Candida species revealed that CHROMagar Candida medium combined with API ID 32C kit yielded the same results (100% compatible) as molecular techniques for the species identification of Candida isolates. Since these phenotypic methods were simple and cost effective when compared to molecular techniques, they should be considered in the identification of Candida species.

摘要

在过去十年中,念珠菌感染的流行病学以及用于治疗这些感染的抗真菌药物发生了重大变化。近年来,念珠菌属已成为重症监护病房患者侵袭性感染的重要病因。本研究的主要目标之一是评估我院分离出的感染性念珠菌属的分子流行病学,并据此为医院感染(HI)控制提供数据。本研究的另一个目的是评估用于将念珠菌分离株鉴定到种水平的传统方法和分子方法的有效性及实际适用性。本研究共纳入了从60例住院患者(29例男性,24例女性;7例为儿童)的各种临床标本中分离出的77株念珠菌菌株。根据美国疾病控制与预防中心(CDC)标准,其中57株(74%)被定义为医院感染病原体。被鉴定为医院感染病原体的最常见念珠菌属是白色念珠菌(22株;38.6%),其次是热带念珠菌(14株;24.6%)、近平滑念珠菌(13株;22.8%)、光滑念珠菌(7株;12.3%)和念珠菌属(1株;1.75%)。确定血流感染(26例;45.6%)和尿路感染(24例;42.1%)是念珠菌属引起的最常见医院感染。此外,检测到血流感染最常见的病原体是近平滑念珠菌(10株;38.5%),尿路感染最常见的病原体是白色念珠菌(12株;50%)对于传统表型方法[芽管形成、玉米粉琼脂中厚垣孢子形成、45°C生长、CHROMagar念珠菌培养基上的菌落特征、通过API ID 32C(法国生物梅里埃公司)系统检测的碳水化合物同化特性]和一些分子技术[使用ITS-1、ITS-3和ITS 4引物的聚合酶链反应(PCR)、PCR-限制性片段长度多态性(RFLP)、用Msp I和Bln I限制性酶切割ITS1-ITS4产物的PCR-RFLP]用于念珠菌属鉴定的优缺点评估显示,CHROMagar念珠菌培养基与API ID 32C试剂盒结合用于念珠菌分离株的种鉴定时产生的结果与分子技术相同(100%兼容)。由于这些表型方法与分子技术相比简单且成本效益高,在念珠菌属鉴定中应予以考虑。

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