導電性中空糸膜を用いた高感度ヘルペスウイルス検査法

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The nucleic acid amplification test (NAT) has been used as a means for detecting viruses in blood. Using this procedure increases the safety of blood products. However, detection is theoretically impossible if the blood is contaminated with only a minimal viral concentration (less than 10^2 PFU/mL). As a result, viral infections continue to occur through the use of blood products. A viral detection method with a higher sensitivity than NAT is required to prevent vial infections such as hepatitis B, hepatitis C and acquired immune deficiency syndrome (AIDS) from occurring. We propose in this paper a viral detection method that uses a hollow-fiber membrane made of polypropylene coated with metal. Using this method, the membrane captures the virus and releases genes in the following manner: Herpes simplex virus type 1 (HSV) in samples was captured by the membrane when the samples were aspirated through the membrane. To lyse the capsid covering the viral gene, the membrane was soaked in a lysis solution containing 1% SDS and 10 mM NaOH. After that, the membrane was set in an electrophoresis bath as a cathode. An electric field was applied to the membrane and HSV genes were released from the membrane because the genes had a negative charge. These genes were amplified by polymerase chain reaction (PCR). PCR products were detected by electrophoresis. In this paper, we succeeded in detecting a virus from HSV suspensions. The end point for detectable concentration of the virus was 30 PFU/mL. This viral concentration level is not possible using the NAT, which has a detection sensitivity of 10^2 PFU/mL. Using the method discussed here, it was possible to detect the viral concentration following aspiration. The sensitivity of this virus detection method is higher than that of NAT. We believe that this method has the possibility of reducing the number of viral infections through blood such as hepatitis B virus and so on.
2003-12-10

導電性中空糸膜を用いた高感度ヘルペスウイルス検査法

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