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Bulletin of the Korean Chemical Society (BKCS)

ISSN 0253-2964(Print)
ISSN 1229-5949(Online)
Volume 29, Number 1
BKCSDE 29(1)
January 20, 2008 

 
Title
Ultra-fast Detection and Differentiation of Mycoplasma haemofelis and Candidatus
M. Haemominutum in Korean Feral Cats by Microchip Electrophoresis
with Programmed Field Strength Gradients
Author
Kailasa S. Kumar, Hee Gu Lee, Dong Jin Yoo, Seong Ho Kang*
Keywords
Chip technology, Feline hemobartonellosis, Polymerase chain reaction, Programmed field strength gradients
Abstract
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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153 - 0
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