NDLI: Establishment and comparison of pulsed-field gel electrophoresis, multiple-locus variable number tandem repeat analysis and automated ribotyping methods for subtyping of Citrobacter strains.

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Establishment and comparison of pulsed-field gel electrophoresis, multiple-locus variable number tandem repeat analysis and automated ribotyping methods for subtyping of Citrobacter strains.

Content Provider	Semantic Scholar
Author	Zhang, Xiao Ai Bai, Xue Mei Ye, Chang Yun Ren, Zhi Hong Xu, Jian Guo
Copyright Year	2012
Abstract	OBJECTIVE To establish and compare the pulsed-field gel electrophoresis (PFGE), multiple-locus variable number tandem repeat analysis (MLVA) and automated ribotyping for subtyping of Citrobacter strains. METHODS PFGE protocol was optimized in terms of plug preparation procedure, restriction enzymes and configuration of electrophoretic parameters. MLVA method was evaluated by finding variable number tandem repeats in two genomes of Citrobacter strains. The ribotyping was performed by using the automated RiboPrinter system. RESULTS We optimized the plug preparation procedure, focused on the cell suspension concentration (turbidity of 2.5 to 3.5), SDS addition (no SDS needed) and lysis time (1 h), and selected the appropriate restriction enzyme (XbaI) and the electrophoretic parameters (1.0 s-20.0 s for 19 h) of PFGE. There was nearly no discriminatory power of MLVA between Citrobacter strains. For 51 Citrobacter strains, automated ribotyping gave a D-value of 0.9945, while PFGE gave a D-value of 0.9969. Both PFGE and automated ribotyping clustered strains from the same sources (with the same species from the same place at the same time identified as the same source) and divided strains from different sources (from different years, places and hosts) into different subtypes. CONCLUSION PFGE protocol established in this paper and automated ribotyping are suitable for application in Citrobacter subtyping.
Starting Page	p292
Ending Page	p293
Page Count	1
File Format	PDF HTM / HTML
Alternate Webpage(s)	http://www.besjournal.com/Articles/Archive/archive/No6/201212/P020121203523243850078.pdf
PubMed reference number	23228835v1
Alternate Webpage(s)	https://doi.org/10.3967/0895-3988.2012.06.007
DOI	10.3967/0895-3988.2012.06.007
Journal	Biomedical and environmental sciences : BES
Volume Number	25
Issue Number	6
Language	English
Access Restriction	Open
Subject Keyword	Citrobacter Common Variable Immunodeficiency DNA Restriction Enzymes Electrophoresis, Gel, Pulsed-Field Gel Electrophoresis (lab technique) Genome Plug (physical object) Ribotyping Subtype (attribute) Suspensions Tandem Repeat Sequences Turbidity Measurement pathologic cytolysis
Content Type	Text
Resource Type	Article

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