tailieunhanh - Báo cáo sinh học: " An inexpensive and rapid diagnostic method of Koi Herpesvirus (KHV) infection by loop-mediated isothermal amplification"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành hóa học dành cho các bạn yêu hóa học tham khảo đề tài: An inexpensive and rapid diagnostic method of Koi Herpesvirus (KHV) infection by loop-mediated isothermal amplification | Virology Journal BioMed Central Methodology Open Access An inexpensive and rapid diagnostic method of Koi Herpesvirus KHV infection by loop-mediated isothermal amplification Hatem Soliman and Mansour El-Matbouli Address Institute of Zoology Fish Biology and Fish Diseases Faculty of Veterinary Medicine University of Munich Germany Email Hatem Soliman - soliman@ Mansour El-Matbouli - el-matbouli@ Corresponding author Published 17 October 2005 Received 27 May 2005 Accepted 17 October 2005 Virology Journal 2005 2 83 doi 1743-422X-2-83 This article is available from http content 2 1 83 2005 Soliman and El-Matbouli licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License http licenses by which permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited. Abstract Background Koi Herpesvirus KHV affects both juvenile and adult common carp and koi and is especially lethal to fry. The high mortalities caused by the disease have had a negative impact on the international koi trade. Different diagnostic techniques have been used to detect KHV including isolation of the virus in cell culture electron microscopy several PCR tests ELISA and in situ hybridisation. All of these methods are time consuming laborious and require specialised equipment. Results A rapid field diagnosis of KHV in common and koi carp was developed using loop-mediated isothermal amplification LAMP . The LAMP reaction rapidly amplified nucleic acid with high specificity and efficiency under isothermal conditions using a simple water bath. Two methods of extracting DNA from host tissue were compared extraction by boiling and by using a commercial extraction kit. A set of six primers - two inner primers two outer primers and two loop primers - was designed from a KHV amplicon. The reaction .