Development of Ultra-rapid Real-Time PCR Method for the detection of Chronic Bee Paralysis Virus

Title
Development of Ultra-rapid Real-Time PCR Method for the detection of Chronic Bee Paralysis Virus
Author(s)
박용하Mi-Sun Yoo[Mi-Sun Yoo]Ji-Na No[Ji-Na No]Van Phu Nguyen[Van Phu Nguyen]Yong-Soo Choi[Yong-Soo Choi]Byoung-Soo Yoon[Byoung-Soo Yoon]
Keywords
Chronic bee paralysis virus; CBPV; Honeybee; Micro PCR; Real-time PCR detection; Ultra-rapid real-time PCR
Issue Date
201010
Publisher
한국양봉학회
Citation
Journal of Apiculture, v.25, no.3, pp.193 - 199
Abstract
Chronic bee paralysis virus (CBPV) is responsible for chronic paralysis, an infectious and contagious disease of adult honey bees (Apis mellifera L.). For the rapid and quantitative detection of CBPV in honeybee, a novel micro PCR-based detection method, termed ultra-rapid real-time PCR, was developed. A specific detection primer set (CBPV Q-2F/2R) was used for amplification of a unique 232bp DNA fragment with a micro-scale chip-based real-time PCR system, GenSpector􄠶 TMC-1000, which has uncommonly fast heating and cooling rates (10°C per second) and a small reaction volume (6μl). Real-time PCR assays using serially diluted DNA templates and artificially contaminated DNA templates showed high sensitivities detection of 10°copies in the both standard assays. This method can detect not only be used for DNA conditions but also be applied to honeybee samples. In the application of ultra-rapid real-time PCR detection of an CBPV-infected honeybee, the minimum detection time was 16 min 8 seconds (30 cycles), including melting temperature analysis. This novel detection method is one of the most rapid real-time PCR-based diagnostic tools and is expected to be applied to the development of rapid detection of various pathogens.
URI
http://hdl.handle.net/YU.REPOSITORY/23415
ISSN
1225-0252
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