Mitsunaga Shigeki, Fujimura Kayoko, Matsumoto Chieko, Shiozawa Rieko, Hirakawa Shinichi, Nakajima Kazunori, Tadokoro Kenji, Juji Takeo
Transfusion Information Department, The Japanese Red Cross Central Blood Center, Tokyo, Japan.
Transfusion. 2002 Jan;42(1):100-6. doi: 10.1046/j.1537-2995.2002.00024.x.
A high-throughput detection system was developed for HBV DNA and HCV RNA.
A combination of real-time detection PCR using an automated system (PRISM 7700, PE Biosystems, Foster City, CA) and automatic viral nucleic acid extraction (BioRobot 9604, Qiagen, Hilden, Germany) was used as the high-throughput detection system. An internal control for HBV DNA detection was also developed.
Testing of 96 samples for HBV and HCV was completed within 5 hours. The sensitivity of this system almost equals that of the manual method using nested PCR. The addition of an internal control for HBV detection did not affect the sensitivity of the method and confirmed the accuracy of results. It was possible to quantify HBV in HBV+ samples that contain more than 500 genome equivalents per mL. We started using this system from June 1999 for testing stored donor and patient samples to analyze cases of posttransfusion hepatitis and identified three HBV+ donations that were implicated in posttransfusion hepatitis B.
The high-throughput detection system is a useful tool for HBV DNA and HCV RNA detection because it enables rapid and reliable testing of a large number of samples.
开发了一种用于检测乙肝病毒DNA（HBV DNA）和丙肝病毒RNA（HCV RNA）的高通量检测系统。
将使用自动化系统（PRISM 7700，PE Biosystems公司，加利福尼亚州福斯特城）的实时检测PCR与自动病毒核酸提取（BioRobot 9604，Qiagen公司，德国希尔德）相结合，用作高通量检测系统。还开发了一种用于HBV DNA检测的内部对照。
96份样本的HBV和HCV检测在5小时内完成。该系统的灵敏度几乎等同于使用巢式PCR的手工方法。添加HBV检测的内部对照不影响该方法的灵敏度，并证实了结果的准确性。对于每毫升含有超过500个基因组当量的HBV阳性样本，可以对HBV进行定量。我们从1999年6月开始使用该系统检测储存的供体和患者样本，以分析输血后肝炎病例，并确定了3份与输血后乙型肝炎有关的HBV阳性献血样本。
高通量检测系统是检测HBV DNA和HCV RNA的有用工具，因为它能够对大量样本进行快速可靠的检测。
