Rapid PCR-based test for identifying Candida albicans by using primers derived from the pH-regulated KER1 gene

被引:13
|
作者
Galan, Amparo
Veses, Veronica
Murgui, Amelia
Casanova, Manuel
Martinez, Jose P. [1 ]
机构
[1] Univ Valencia, Fac Farm, Dept Microbiol & Ecol, E-46100 Burjassot, Valencia, Spain
[2] Univ Valencia, Fac Farm, Dept Bioquim & Biol Mol, E-46100 Burjassot, Valencia, Spain
[3] Banco Lineas Celulares, Ctr Invest Principe Felipe, Valencia, Spain
[4] Univ Aberdeen, Sch Med Sci, Dept Mol & Cell Biol, Aberdeen AB9 1FX, Scotland
关键词
Candida albicans; molecular identification; PCR; KER1; gene; DNA extraction;
D O I
10.1111/j.1567-1364.2006.00114.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
A PCR-based method in combination with a simple, reliable and inexpensive DNA extraction procedure for rapid detection of Candida albicans clinical isolates is described here. The extraction protocol is based on a combination of chemical (NaOH and detergents) and physical (boiling) treatments, thus avoiding many of the problems inherent in the currently available DNA extraction protocols (basically the use of expensive and/or toxic chemical reagents), and may be useful for daily clinical routine. The PCR-based system described here uses a single pair of primers (SC1F and SC1R) deduced from the C. albicans-specific KER1 gene sequence. These primers amplify a 670-bp fragment of the KER1 gene. All the clinical C. albicans isolates generated the expected 670-bp amplicon. Other non-albicans Candida species, including the azole-resistant C. krusei and C. glabrata, and the very closely related C. dubliniensis, failed to amplify any DNA fragment. The PCR results reported here suggest that amplification with SC1F and SC1R primers is species specific and, consequently, may be useful for specifically identifying C. albicans strains.
引用
收藏
页码:1094 / 1100
页数:7
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  • [1] Rapid PCR test for discriminating between Candida albicans and Candida dubliniensis isolates using primers derived from the pH-regulated PHR1 and PHR2 genes of C-albicans
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