Comparison on genotyping of Candida parapsilosis by polymorphic microsatellite marker technology and repetitive sequence-based PCR

doi:10.3969/j.issn.1673-8640.2015.10.013

Abstract

Abstract: Objective

To evaluate the application of polymorphic microsatellite marker(PMM)technology and repetitive sequence-based polymerase chain reaction (PCR) for genotyping of Candida parapsilosis.

Methods

From March 2012 to November 2013, 55 Candida parapsilosis isolates were collected from 5 hospitals in Shanghai, which were further identified as Candida parapsilosis sensu stricto (43 isolates) by amplifying and sequencing with internal transcribed sequence(ITS). All the isolates were genotyped by repetitive sequence-based PCR and PMM technology. Finally, the results and efficacy of these 2 genotyping methods were compared.

Results

The 43 Candida parapsilosis sensu stricto isolates had only 1 genotype by repetitive sequence-based PCR. PMM technology generated 22 genotypes. The major genotypes were Type Ⅰ (10 isolates) and Type Ⅱ (6 isolates), whereas Type Ⅲ-Ⅶ were observed in 2 or 3 isolates. Another 15 genotypes were observed in only 1 isolate.

Conclusions

PMM technology is an effective genotyping method for Candida parapsilosis sensu stricto with high resolution and repeatability, which could be used to determine the micro-evolutionary variations. However, repetitive sequence-based PCR is not suitable for the genotyping of Candida parapsilosis sensu stricto.

Key words: Candida parapsilosis, Genotype, Polymorphic microsatellite marker technology, Repetitive sequence-based polymerase chain reaction

CLC Number:

R379.4

LI Zhen, DONG Danfeng, ZHANG Lihua, JIANG Cen, PENG Yibing.. Comparison on genotyping of Candida parapsilosis by polymorphic microsatellite marker technology and repetitive sequence-based PCR[J]. Laboratory Medicine, 2015, 30(10): 1017-1020.

Figures/Tables 3

References 12

[1]	TROFA D,GACSER A,NOSANCHUK JD.Candida parapsilosis,an emerging fungal pathogen[J]. Clin Microbiol Rev,2008,21(4):606-625.
[2]	TRAGIANNIDIS A,FEGELER W,RELLENSMANN G,et al.Candidaemia in a European Paediatric University Hospital: a 10-year observational study[J]. Clin Microbiol Infect,2012,18(2):E27-E30.
[3]	SABINO R, SAMPAIO P,ROSADO L,et al.New polymorphic microsatellite markers able to distinguish among Candida parapsilosis sensu stricto isolates[J]. J Clin Microbiol,2010,48(5):1677-1682.
[4]	TAVANTI A,DAVIDSON AD,GOW NA,et al.Candida orthopsilosis and Candida metapsilosis spp. nov. to replace Candida parapsilosis groups Ⅱ and Ⅲ[J]. J Clin Microbiol,2005,43(1):284-292.
[5]	REDKAR RJ,DUBEÉ MP,MCCLESKEY FK,et al.DNA fingerprinting of Candida rugosa via repetitive sequence-based PCR[J]. J Clin Microbiol ,1996,34(7):1677-1681.
[6]	VAN ASBECK EC, CLEMONS KV, MARKHAM AN, et al.Correlation of restriction fragment length polymorphism genotyping with internal transcribed spacer sequence, randomly amplified polymorphic DNA and multilocus sequence groupings for Candida parapsilosis[J]. Mycoses, 2009, 52(6): 493-498.
[7]	ZANCOPE-OLIVEIRA R M, JAMES M J, DEROSSI AP, et al. Strain characterization of Candida parapsilosis fungemia by molecular typing methods[J]. Eur J Clin Microbiol Infect Dis, 2000, 19(7): 514-520.
[8]	LASKER BA,BUTLER G,LOTT TJ.Molecular genotyping of Candida parapsilosis group I clinical isolates by analysis of polymorphic microsatellite markers[J]. J Clin Microbiol,2006,44(3):750-759.
[9]	姚冬婷,应春妹,郑冰. 微卫星多态性和多位点序列分析技术在光滑念珠菌基因分型中的应用评估[J].检验医学,2014,29(6):593-596.
[10]	江岑,董丹凤,俞焙秦, 等. 重复序列PCR 与多位点分型技术在热带假丝酵母菌基因分型中的比较[J]. 检验医学,2012,27(11): 917-920.
[11]	董丹凤,江岑,俞焙秦, 等. 比较重复序列 PCR 与多位点序列分型技术用于光滑假丝酵母菌的基因分型检测[J]. 中华检验医学杂志, 2011,34(9): 810-813.
[12]	魏冰,刘锦燕,史册, 等. 重复序列PCR 与多位点分型技术在白假丝酵母菌基因分型中的比较[J]. 上海交通大学学报:医学版,2013,33(11): 1445-1448.

PMM技术			重复序列PCR
分型编号		分型结果	菌株数(株)
Ⅰ	232/272	10	A
Ⅱ	289/289	6	A
Ⅲ	297/297	3	A
Ⅳ	264/264	2	A
Ⅴ	269/269	2	A
Ⅵ	267/285	3	A
Ⅶ	267/276	2	A

PMM技术			重复序列PCR
分型编号		分型结果	菌株数(株)
Ⅰ	232/272	10	A
Ⅱ	289/289	6	A
Ⅲ	297/297	3	A
Ⅳ	264/264	2	A
Ⅴ	269/269	2	A
Ⅵ	267/285	3	A
Ⅶ	267/276	2	A

菌株	PMM	重复序列PCR
CO1	169/187	B
CO2	169/187	B
CO3	169/187	B
CO4	169/187	B
CO5	169/187	B
CO6	169/187	B
CO7	169/187	B
CM1	240/240	C
CM2	232/272	D
CM3	-	E
CM4	-	C
CM5	-	D

菌株	PMM	重复序列PCR
CO1	169/187	B
CO2	169/187	B
CO3	169/187	B
CO4	169/187	B
CO5	169/187	B
CO6	169/187	B
CO7	169/187	B
CM1	240/240	C
CM2	232/272	D
CM3	-	E
CM4	-	C
CM5	-	D

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