Evaluation of nested and real-time PCR assays in the diagnosis of candidaemia

被引:31
作者
Khlif, M. [2 ]
Mary, C. [1 ]
Sellami, H. [2 ]
Sellami, A. [2 ]
Dumon, H. [1 ]
Ayadi, A. [2 ]
Ranque, S. [1 ]
机构
[1] Hop Enfants La Timone, Lab Parasitol Mycol, F-13385 Marseille 05, France
[2] Univ Sfax, Fac Med, Lab Biol Mol Parasitaire & Fong, Sfax, Tunisia
关键词
Blood culture; Candida; diagnosis; nested PCR; real-time PCR; POLYMERASE-CHAIN-REACTION; RAPID IDENTIFICATION; WHOLE-BLOOD; FUNGAL-INFECTIONS; ALBICANS DNA; ASPERGILLUS; PATHOGENS; SERUM; AMPLIFICATION; CANDIDEMIA;
D O I
10.1111/j.1469-0691.2009.02762.x
中图分类号
R51 [传染病];
学科分类号
100401 ;
摘要
P>Polymerase chain reaction (PCR) assays have a very low theoretical detection threshold and are therefore advocated for the diagnosis of fungaemia. However, their effectiveness in this respect remains to be assessed. This study compared real-time PCR (Can-G) and nested PCR assays with blood culture for the diagnosis of Candida spp. bloodstream infections. A total of 200 clinical blood samples obtained from 110 patients at risk for developing a systemic fungal infection, hospitalized in the University Hospital of Sfax (Tunisia), were submitted to testing by culture, nested PCR and real-time PCR. Blood culture was positive in 36 patients. When compared with culture, the Can-G assay (81% sensitivity, 96% specificity) performed better than the nested PCR assay (86% sensitivity, 54% specificity). The real-time PCR assay, which avoids both the contamination hazard with amplicons that may cause false-positive results and the use of time-consuming post-PCR steps, appears more suitable than the nested PCR assay for the laboratory diagnosis of Candida spp. bloodstream infections. In this study, real-time PCR did not enhance the diagnostic sensitivity for Candida spp. bloodstream infections compared with conventional blood culture; however, it may lead to earlier implementation of an adequately targeted antifungal treatment.
引用
收藏
页码:656 / 661
页数:6
相关论文
共 32 条
[1]   Comparative evaluation of (1,3)-β-D-glucan, mannan and anti-mannan antibodies, and Candida species-specific snPCR in patients with candidemia [J].
Alam, Fasahat F. ;
Mustafa, Abu S. ;
Khan, Zia U. .
BMC INFECTIOUS DISEASES, 2007, 7 (1)
[2]  
[Anonymous], 1990, PCR Protocols. A Guide to Methods and Applications
[3]  
Badiee P, 2007, EXP CLIN TRANSPLANT, V5, P624
[4]   Serum is more suitable than whole blood for diagnosis of systemic candidiasis by nested PCR [J].
Bougnoux, ME ;
Dupont, C ;
Mateo, J ;
Saulnier, P ;
Faivre, V ;
Payen, D ;
Nicolas-Chanoine, MH .
JOURNAL OF CLINICAL MICROBIOLOGY, 1999, 37 (04) :925-930
[5]  
BUCHMAN TG, 1990, SURGERY, V108, P338
[6]   Rapid identification of yeasts in positive blood cultures by a multiplex PCR method [J].
Chang, HC ;
Leaw, SN ;
Huang, AH ;
Wuk, TL ;
Chang, TC .
JOURNAL OF CLINICAL MICROBIOLOGY, 2001, 39 (10) :3466-3471
[7]   Polymorphic internal transcribed spacer region 1 DNA sequences identify medically important yeasts [J].
Chen, YC ;
Eisner, JD ;
Kattar, MM ;
Rassoulian-Barrett, SL ;
Lafe, K ;
Bui, U ;
Limaye, AP ;
Cookson, BT .
JOURNAL OF CLINICAL MICROBIOLOGY, 2001, 39 (11) :4042-4051
[8]   DETECTION OF CANDIDA-ALBICANS DNA IN SERUM BY POLYMERASE CHAIN-REACTION [J].
CHRYSSANTHOU, E ;
ANDERSSON, B ;
PETRINI, B ;
LOFDAHL, S ;
TOLLEMAR, J .
SCANDINAVIAN JOURNAL OF INFECTIOUS DISEASES, 1994, 26 (04) :479-485
[9]   Detection and identification of fungal pathogens in blood by using molecular probes [J].
Einsele, H ;
Hebart, H ;
Roller, G ;
Loffler, J ;
Rothenhofer, I ;
Muller, CA ;
Bowden, RA ;
vanBurik, JA ;
Engelhard, D ;
Kanz, L ;
Schumacher, U .
JOURNAL OF CLINICAL MICROBIOLOGY, 1997, 35 (06) :1353-1360
[10]   Multiplex PCR using internal transcribed spacer 1 and 2 regions for rapid detection and identification of yeast strains [J].
Fujita, SI ;
Senda, Y ;
Nakaguchi, S ;
Hashimoto, T .
JOURNAL OF CLINICAL MICROBIOLOGY, 2001, 39 (10) :3617-3622