Department of Safety Research on Blood and Biologics, Department of Virology, Medicine, National Institute of Infectious Diseases, Tokyo, Japan; Department of Pathology, Yamaguchi University Graduate School of Medicine, Yamaguchi, Japan; Department of Veterinary Medicine, Rakuno Gakuen University, Hokkaido, Japan; Osaka Red Cross Blood Center, Osaka, Japan; Nihon Parkerizing Hiroshima Works Co. Ltd, Hiroshima, Japan.
Transfusion. 2013 Oct;53(10 Pt 2):2545-55. doi: 10.1111/trf.12193. Epub 2013 Apr 17.
The risk of transferring blood-borne infections during transfusion is continually increasing because of newly emerging and reemerging viruses. Development of a rapid screening method for emerging viruses that might be transmitted by transfusion is required to eliminate such pathogens during blood donor screening. Owing to increased use of human materials in organ transplants and cell therapy, the risk of donor-transmitted viral infections is also increasing. Although nucleic acid amplification technology (NAT) is dedicated to blood screening, a small, convenient detection system is needed at the laboratory and hospital level.
We developed a new pathogen detection system that can detect multiple viruses simultaneously, using originally designed degenerate polymerase chain reaction primers to amplify a wide range of viral genotypes. Amplified samples were identified using a DNA microarray of pathogen-specific probes.
We detected very low copy numbers of multiple subtypes of viruses, such as human hepatitis C virus (HCV), human hepatitis B virus (HBV), human parvovirus B19 (PVB19), and West Nile virus (WNV), using a single plate. We also detected all genotypes of human immunodeficiency virus (HIV) but sensitivity was less than for the other viruses.
We developed a microarray assay using novel primers for detection of a wide range of multiple pathogens and subtypes. Our NAT system was accurate and reliable for detection of HIV, HBV, HCV, PVB19, and WNV, with respect to specificity, sensitivity, and genotype inclusivity. Our system could be customized and extended for emerging pathogens and is suitable as a future NAT system.
由于新出现和重新出现的病毒，输血过程中血液传播感染的风险不断增加。需要开发一种快速筛选方法来检测可能通过输血传播的新兴病毒，以便在献血者筛查过程中消除这些病原体。由于在器官移植和细胞治疗中越来越多地使用人体材料，供体传播的病毒感染风险也在增加。尽管核酸扩增技术 (NAT) 专门用于血液筛查，但在实验室和医院层面需要一种小型、方便的检测系统。
我们开发了一种新的病原体检测系统，该系统使用最初设计的简并聚合酶链反应引物，可以同时检测多种病毒，以扩增广泛的病毒基因型。使用病原体特异性探针的 DNA 微阵列来鉴定扩增的样品。
我们使用单个平板检测到了多种病毒亚型的极低拷贝数，例如人类丙型肝炎病毒 (HCV)、人类乙型肝炎病毒 (HBV)、人类细小病毒 B19 (PVB19) 和西尼罗河病毒 (WNV)。我们还检测到了所有基因型的人类免疫缺陷病毒 (HIV)，但敏感性低于其他病毒。
我们开发了一种使用新型引物的微阵列检测方法，用于检测广泛的多种病原体和亚型。我们的 NAT 系统在特异性、敏感性和基因型包容性方面对 HIV、HBV、HCV、PVB19 和 WNV 的检测均准确可靠。我们的系统可以定制和扩展以检测新出现的病原体，适合作为未来的 NAT 系统。
