Multilocus sequence typing for Candida albicans isolates from candidemic patients: comparison with Southern blot hybridization and pulsed-field gel electrophoresis analysis.

Youn Myoung, Jong Hee Shin, Jin Sol Lee, Soo Hyun Kim, Myung Geun Shin, Soon Pal Suh, Dong Wook Ryang
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引用次数: 12

Abstract

Background: We evaluated the efficacy of multilocus sequence typing (MLST) for assessing the genetic relationship among Candida albicans isolates from patients with candidemia in a hospital setting.

Methods: A total of 45 C. albicans isolates from 21 patients with candidemia were analyzed. The MLST results were compared with results obtained by Southern blot hybridization (C1 fingerprinting) and pulsed-field gel electrophoresis (PFGE). PFGE analysis included karyotyping and restriction endonuclease analysis of genomic DNAs using BssHII (REAG-B) and SfiI (REAG-S).

Results: The 45 isolates yielded 20 unique diploid sequence types (DSTs) by MLST, as well as 12 karyotypes, 15 REAG-B patterns, 13 REAG-S patterns, and 14 C1 fingerprinting types. Microevolution among intra-individual isolates was detected in 6, 5, 3, 5, and 7 sets of isolates by MLST (1 or 2 allelic differences), REAG-B, REAG-S, C1 fingerprinting, and a combination of all methods, respectively. Among 20 DSTs, 17 were unique, and 3 were found in more than 1 patient. The results of 2 DSTs obtained from 9 patient isolates were in agreement with REAG and C1 fingerprinting patterns. However, the remaining DST, which was shared by 2 patient isolates, showed 2 different PFGE and C1 fingerprinting patterns. In addition, 3 sets of isolates from different patients, which differed in only 1 or 2 alleles by MLST, also exhibited different PFGE or C1 fingerprinting patterns.

Conclusions: MLST is highly discriminating among C. albicans isolates, but it may have some limitations in typing isolates from different patients, which may necessitate additional analysis using other techniques.

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白色念珠菌分离株的多位点序列分型:与Southern blot杂交和脉冲场凝胶电泳分析的比较。
背景:我们评估了多位点序列分型(MLST)用于评估医院念珠菌患者分离的白色念珠菌之间的遗传关系的有效性。方法:对21例念珠菌病患者的45株白色念珠菌进行分析。将MLST结果与Southern blot杂交(C1指纹)和脉冲场凝胶电泳(PFGE)结果进行比较。PFGE分析包括使用BssHII (REAG-B)和SfiI (REAG-S)对基因组dna进行核型和限制性内切酶分析。结果:45株分离菌株经MLST鉴定得到20个独特的二倍体序列型(DSTs), 12个核型,15个REAG-B型,13个REAG-S型,14个C1指纹型。分别采用MLST(1个或2个等位基因差异)、REAG-B、REAG-S、C1指纹图谱和多种方法联合检测6、5、3、5和7组分离物的个体内微进化。20例DSTs中,17例为独特的,3例在1例以上患者中发现。从9株患者分离株中获得的2个DSTs结果与REAG和C1指纹图谱一致。然而,其余的DST,由2例患者分离株共享,显示2种不同的PFGE和C1指纹图谱。此外,来自不同患者的3组分离株,在MLST上仅存在1或2个等位基因的差异,也表现出不同的PFGE或C1指纹图谱。结论:MLST在白色念珠菌分离株中具有高度的鉴别性,但在不同患者分离株分型时可能存在一定的局限性,这可能需要使用其他技术进行额外的分析。
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来源期刊
Korean Journal of Laboratory Medicine
Korean Journal of Laboratory Medicine 医学-医学实验技术
自引率
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1
审稿时长
>12 weeks
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