Ultra-fast detection and differentiation of Mycoplasma haemofelis and Candidatus M. haemominutum in Korean feral cats by microchip electrophoresis with programmed field strength gradients

被引：0
作者：
Kumar, Kailasa S. [1 ]
Hee, Gu Lee [2 ]
Dong, Jin Yoo [3 ]
Seong, Ho Kang [1 ]
机构：
[1] Department of Chemistry, Basic Science Research Institute, Chonbuk National University
[2] Cellomics Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)
[3] Department of Chemistry, Seonam University
来源：
Bulletin of the Korean Chemical Society | 2008年 / 29卷 / 01期
关键词：
Chip technology; Feline hemobartonellosis; Polymerase chain reaction; Programmed field strength gradients;
D O I：
10.5012/bkcs.2008.29.1.153
中图分类号：
学科分类号：
摘要：
A microchip-based capillary gel electrophoresis (MCGE) technique was developed for the ultra-fast detection and differentiation of Candidatus Mycoplasma haemominutum (Candidatus M. haemominutum, California strain) and Mycoplasma haemofelis (M. haemofelis, Ohio strain) in Korean feral cats through the application of programmed field strength gradients (PFSG) in a conventional glass double-T microchip. The effects of the poly (ethyleneoxide) (PEO) concentration and electric field strength on the separation of DNA fragments were investigated. The PCR-amplified products of Candidatus M. haemominutum (202-bp) and M. haemofelis (273-bp) were analyzed by MCGE within 75 s under a constant applied electric field of 117.6 V/cm and a sieving matrix of 0.3% PEO (Mr 8 000 000). When the PFSG was applied, MCGE analysis generated results 6.8-times faster without any loss of resolution or reproducibility. The MCGE-PFSG technique was also applied to eleven samples selected randomly from 33 positive samples. The samples were detected and differentiated within 11 s. The analysis time of the MCGE-PFSG technique was approximately 980-times faster than that using conventional slab gel electrophoresis.
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页码：153 / 158
页数：5
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