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[通过26S rDNA D1/D2结构域的单链构象多态性分析快速鉴定临床酵母菌种]

[Rapid identification of clinical yeast species by single-strand conformation polymorphism analysis of 26S rDNA D1/D2 domain].

作者信息

Li Juan, Bai Fengyan

机构信息

State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, China CDC, Beijing 102206, China.

出版信息

Wei Sheng Wu Xue Bao. 2009 Aug;49(8):1011-7.

Abstract

OBJECTIVE

To investigate the reliability of single-strand conformation polymorphism (SSCP) analysis of 26S rDNA D1/D2 domain for rapid identification of clinical yeast species and to examine the distribution of the yeast species in clinical strains from Beijing.

METHODS

Type and authentic strains of five common pathogenic yeast species were used as references. Approximately 260 yeast strains with diversified clinical origins were collected from four hospitals located in Beijing. The 26S rDNA D1/D2 domain of each strain was amplified by PCR and subjected to SSCP or sequence analysis.

RESULTS

SSCP analysis showed that the Candida strains with slight sequence differences in the D1/D2 domain could be effectively detected. The common pathogenic Candida species, including C. albicans, C. parapsilosis, C. tropicalis, C. glabrata and C. krusei, were clearly distinguished from each other by their SSCP patterns of PCR amplified D1/D2 domain products. Twenty species belonging to 10 genera were identified from the approximately 260 clinical yeast strains based on SSCP pattern comparison for the common species and D1/D2 sequence analysis for the uncommon species. The dominant species and their frequencies were: C. albicans (57.7%), C. parapsilosis (10.0%), C. tropicalis (9.2%), C. glabrata (6.7%) and C. krusei (5.8%).

CONCLUSION

The results indicated that PCR-SSCP analysis of D1/D2 is a powerful approach for rapid species identification of clinical yeast strains. The most common clinical yeast species was C. albicans in Beijing but the increasing trend of non-albicans Candida species was observed.

摘要

目的

研究26S rDNA D1/D2区域单链构象多态性(SSCP)分析用于快速鉴定临床酵母菌种的可靠性,并检测北京临床菌株中酵母菌种的分布情况。

方法

以5种常见致病性酵母菌种的标准菌株和 authentic 菌株作为对照。从北京的4家医院收集了约260株临床来源多样的酵母菌株。通过PCR扩增各菌株的26S rDNA D1/D2区域,并进行SSCP或序列分析。

结果

SSCP分析表明,能有效检测出D1/D2区域序列存在微小差异的念珠菌菌株。常见致病性念珠菌菌种,包括白色念珠菌、近平滑念珠菌、热带念珠菌、光滑念珠菌和克柔念珠菌,通过其PCR扩增D1/D2区域产物的SSCP图谱能明显区分。基于常见菌种的SSCP图谱比较和罕见菌种的D1/D2序列分析,从约260株临床酵母菌株中鉴定出10个属的20个菌种。优势菌种及其频率为:白色念珠菌(57.7%)、近平滑念珠菌(10.0%)、热带念珠菌(9.2%)、光滑念珠菌(6.7%)和克柔念珠菌(5.8%)。

结论

结果表明,D1/D2区域的PCR-SSCP分析是快速鉴定临床酵母菌株菌种的有效方法。北京临床最常见的酵母菌种是白色念珠菌,但非白色念珠菌菌种呈上升趋势。

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