Fournier-Wirth C, Coste J
Transfusion Transmitted Infectious Diseases R&D Unit, EFS Pyrénées Méditerranée, Montpellier, France.
Dev Biol (Basel). 2007;127:61-70.
Until the late 1990s, mandatory blood screening for transmissible infectious agents depended entirely on antigen/antibody-based detection assays. The recent emergence of Nucleic acid Amplification Technologies (NAT) has revolutionised viral diagnosis, not only by increasing the level of sensitivity but also by facilitating the detection of several viruses in parallel by multiplexing specific primers. In more complex biological situations, when a broad spectrum of pathogens must be screened, the limitations of these first generation technologies became apparent. High throughput systems, such as DNA Arrays, permit a conceptually new approach. These miniaturised micro systems allow the detection of hundreds of different targets simultaneously, inducing a dramatic decrease in reagent consumption, a reduction in the number of confirmation tests and a simplification of data interpretation. However, the systems currently available require additional instrumentation and reagents for sample preparation and target amplification prior to detection on the DNA array. A major challenge in the area of DNA detection is the development of methods that do not rely on target amplification systems. Likewise, the advances of protein microarrays have lagged because of poor stability of proteins, complex coupling chemistry and weak detection signals. Emerging technologies like Biosensors and nano-particle based DNA or Protein Bio-Barcode Amplification Assays are promising diagnostic tools for a wide range of clinical applications, including blood donation screening.
直到20世纪90年代末,针对可传播感染因子的强制性血液筛查完全依赖基于抗原/抗体的检测方法。核酸扩增技术(NAT)的出现彻底改变了病毒诊断,不仅提高了灵敏度,还通过复用特异性引物实现了多种病毒的同时检测。在更复杂的生物学情况下,当需要筛查多种病原体时,这些第一代技术的局限性就变得明显了。高通量系统,如DNA阵列,提供了一种全新的概念性方法。这些微型化的微系统能够同时检测数百种不同的靶点,显著减少试剂消耗,减少确认试验次数,并简化数据解读。然而,目前可用的系统在DNA阵列检测之前,需要额外的仪器和试剂进行样品制备和靶点扩增。DNA检测领域的一个主要挑战是开发不依赖靶点扩增系统的方法。同样,由于蛋白质稳定性差、偶联化学复杂以及检测信号微弱,蛋白质微阵列的发展也较为滞后。生物传感器和基于纳米颗粒的DNA或蛋白质生物条形码扩增检测等新兴技术,对于包括献血筛查在内的广泛临床应用而言,是很有前景的诊断工具。