Abstract
The Japanese Red Cross Society is using nucleic acid amplification testing (NAT) to secure the microbiological safety of blood products. The serum has individual difference because the blood is collected from many humans located in various parts of the country. The association reported that the results of NAT were influenced by slight differences in blood components. We designed a viral detection method consisting of a metal-coated hollow fiber (MCHF)-membrane and loop-mediated isothermal amplification (LAMP). This detection method was highly sensitive and much quicker than NAT. In this study, we used two kinds of simulated serum samples that used the bovine serums of different lot numbers and reviewed stability for slight differences in blood components in terms of the designed viral detection. We examined the influence of individual differences on only LAMP for bovine serum of different lot numbers as the first step. LAMP couldn't detect the virus in nay of the simulated serum samples. As the next step, we tested the influence of serum individual differences on viral detection consisting of MCHF-membrane and LAMP. As a result, this viral detection method was able to detect herpes simplex virus type 1 from simulated serum samples containing more than 10 PFU/mL of virus without influence from individual differences. MCHF-membrane possesses an isolation function, in which it captures HSV-1 of more than 100 nm in membrane pore size and passes the bovine serum origin protein, etcetera of less than 100 nm in pore size. Therefore, even if simulated serum samples have individual differences, components in the serum are removed. Gene amplification such as LAMP after viral capture and gene isolation using MCHF-membrane in this viral detection method was uninfluenced by serum individual differences. This viral detection method is stable and not influenced by serum individual differences, and is thus an epoch-making method.
