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通过聚合酶链反应鉴定耳念珠菌并采用结晶紫法评估生物膜形成。

Identification of Candida auris by PCR and assessment of biofilm formation by crystal violet assay.

作者信息

Singh Parul, Srivastava Smriti, Malhotra Rajesh, Mathur Purva

机构信息

Department of Microbiology, Trauma Centre, AIIMS, New Delhi, India.

Department of Orthopaedics & Chief, Trauma Centre, AIIMS, New Delhi, India.

出版信息

Indian J Med Microbiol. 2023 Nov-Dec;46:100421. doi: 10.1016/j.ijmmb.2023.100421. Epub 2023 Jul 5.

DOI:10.1016/j.ijmmb.2023.100421
PMID:37945115
Abstract

INTRODUCTION

Candida auris is a notorious pathogen capable of forming biofilms on devices as well as host tissues, often culminating in infections. We evaluated characteristics of infections and the methods to diagnose C. auris over a period of three years in a tertiary care hospital.

METHODS

Patients admitted between 2018 and 2020, who had candidemia due to C. auris were included in the study. Identification was performed using HiCrome™ Candida Differential Agar, Vitek 2 (BioMérieux, Inc., Marcy-l'Etoile, France) and MALDI-TOF, Vitek-MS. Identification was confirmed by detection of rDNA region covering part of 5.8S, entire of ITS2, and part of 28S by polymerase chain reaction (PCR). Biofilm formation was assessed by crystal violet staining.

RESULTS

Presence of central line and broad spectrum antimicrobials were noted in all patients whereas total parenteral nutrition was given in 82.1% of these patients. Identification by Vitek2 v8.1 correlated with MALDI-TOF MS. PCR products of length 163 ​bp were obtained in all isolates as visualized by agarose gel electrophoresis. The biofilm quantity measured as A of the twenty-eight C. auris isolates ranged from 0.16 to 0.80 compared to C. albicans.

CONCLUSIONS

C. auris can be identified by PCR targeting specific rDNA region. Biofilm formation and quantification can be achieved by growing C. auris isolates in Mueller-Hinton broth over a duration of 48 ​h.

摘要

引言

耳念珠菌是一种臭名昭著的病原体,能够在医疗器械以及宿主组织上形成生物膜,常常导致感染。我们评估了一家三级护理医院在三年时间里耳念珠菌感染的特征及诊断方法。

方法

纳入2018年至2020年间因耳念珠菌导致念珠菌血症而入院的患者。使用HiCrome™念珠菌鉴别琼脂、Vitek 2(法国马西-埃图瓦勒生物梅里埃公司)和基质辅助激光解吸电离飞行时间质谱(MALDI-TOF)、Vitek-MS进行鉴定。通过聚合酶链反应(PCR)检测覆盖部分5.8S、整个ITS2和部分28S的rDNA区域来确认鉴定结果。通过结晶紫染色评估生物膜形成情况。

结果

所有患者均有中心静脉置管并使用了广谱抗菌药物,82.1%的患者接受了全胃肠外营养。Vitek2 v8.1的鉴定结果与MALDI-TOF质谱法相关。通过琼脂糖凝胶电泳可见,所有分离株均获得了长度为163 bp的PCR产物。与白色念珠菌相比,28株耳念珠菌分离株的生物膜量(以A值衡量)在0.16至0.80之间。

结论

可通过针对特定rDNA区域的PCR来鉴定耳念珠菌。在穆勒-欣顿肉汤中培养耳念珠菌分离株48小时,可实现生物膜的形成及定量分析。

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