Myoung Youn, Shin Jong Hee, Lee Jin Sol, Kim Soo Hyun, Shin Myung Geun, Suh Soon Pal, Ryang Dong Wook
Department of Laboratory Medicine, Chonnam National University Medical School, Gwangju, Korea.
Korean J Lab Med. 2011 Apr;31(2):107-14. doi: 10.3343/kjlm.2011.31.2.107.
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.
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).
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.
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.
我们评估了多位点序列分型(MLST)在评估医院环境中念珠菌血症患者分离出的白色念珠菌菌株之间遗传关系的有效性。
对来自21例念珠菌血症患者的45株白色念珠菌进行分析。将MLST结果与Southern印迹杂交(C1指纹图谱)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进行核型分析和限制性内切酶分析。
45株菌株通过MLST产生了20种独特的二倍体序列类型(DST),以及12种核型、15种REAG-B模式、13种REAG-S模式和14种C1指纹图谱类型。通过MLST(1或2个等位基因差异)、REAG-B、REAG-S、C1指纹图谱以及所有方法的组合,分别在6、5、3、5和7组菌株中检测到个体内菌株间的微进化。在20种DST中,17种是独特的,3种在不止1例患者中发现。从9例患者分离株获得的2种DST结果与REAG和C1指纹图谱模式一致。然而,另外2例患者分离株共有的DST显示出2种不同的PFGE和C1指纹图谱模式。此外,3组来自不同患者的分离株,通过MLST仅在1或2个等位基因上存在差异,也表现出不同的PFGE或C1指纹图谱模式。
MLST在白色念珠菌分离株中具有高度鉴别力,但在对不同患者的分离株进行分型时可能存在一些局限性,这可能需要使用其他技术进行额外分析。