National Agriculture and Food Research Organization (NARO) and Takara Bio Co., Ltd. have jointly developed a new simple pre-processing method of blood samples for viral gene detection by real-time PCR. This makes it possible to quickly extract nucleic acids from viruses in blood with only a short heat treatment and centrifugation, and to easily remove components that inhibit PCR reactions. By applying this method to the "Classical Swine Fever Virus (CSFV)/African Swine Fever Virus (ASFV) Differential Test Method" used at prefectural animal hygiene centers and the "CSFV Field Strain/Vaccine Strain Differential Test Method" for wild boars, it is expected to largely contribute to prompt implementation of counter measures against these devastating animal diseases through rapid and reliable detection of both causative viruses in various specimens such as blood, sera, and tissue homogenates. A pre-processing reagent suitable for this technology will be released by Takara Bio Inc. on June 28, 2023.
Overview
CSF, which was reported in domestic pig farms in 2018 for the first time in 26 years, continues to affect pigs in domestic farms and also spreading in wild boars. On the other hand, whilst ASF was not yet reported in Japan, it is necessary to be vigilant against the invasion since it is currently rampant on Asian countries excluding Japan and Taiwan. Although CSF and ASF are different diseases caused by distinct viruses, the clinical symptoms are very similar, so molecular diagnosis aimed at detecting each virus is necessary not only to detect but also to differentiate both diseases.
NARO and Takara Bio Inc. have jointly developed a new method for preparing nucleic acid samples suitable for real-time PCR testing from blood, etc. more easily and quickly than ever before. We have already developed a test method for rapidly and accurately distinguishing and diagnosing CSF and ASF and with the outbreak of CSF in Japan, a new molecular testing method to differentiate the virus strains prevalent in the field and the ones contained in vaccines which helps to enhance immunity of pigs and wild boars. These test methods are used for the field screening of CSF and ASF affected animals by prefectural animal hygiene centers. However, these tests often require the use of blood or poorly preserved specimens that are not suitable for virus detection by real-time PCR, and it takes time to prepare nucleic acid samples that can be used for testing. The newly developed pre-processing method enables the preparation of test samples for the real time PCR simply by performing a short heat treatment and a low-speed centrifugation after mixing the sample and reagent.
By applying this method to the above-mentioned "CSFV/ASFV Differential Test Method" and "CSFV Field Strain/Vaccine Strain Differential Test Method" for wild boars, it is expected to contribute to prompt detection of both major infectious diseases of pigs and wild boars and enable early implementation of counter measures. A pre-processing reagent suitable for this technology will be released by Takara Bio Inc. on June 28, 2023.
