2005年以来，二代测序平台和测序技术越来越普及，被广泛用于新病毒和未知病原体的发现，从未经培养的复杂样本中筛查病毒类病原体需要更多的前期处理措施。我们围绕高通量测序所涉及到的测序样本预处理方法和扩增方法做简要总结：样品类型分为无菌样本和开放样本；病原体类型包括病毒、细菌、真菌等；背景核酸去除的常用方法包括物理分离、核酸酶消化和几种rRNA去除的方法；常用的微量样本非特异性扩增方法包括随机引物PCR、SISPA技术、锚定随机引物PCR、RCA技术、MDA技术和NASBA技术。%Since 2005, the platforms and technology of next generation sequencing were becoming increasingly popular and were widely used to discovery novel viruses and unknown pathogens. More pretreatment measures are required to discovery the pathogens such as virus, especially from a complex sample without culture. We summa⁃rized the pretreatment method and the amplification method of the samples involved in high-throughput sequenc⁃ing. The samples are classified into sterile samples and open samples by the origin type and are classified into vi⁃ruses, bacteria, fungi, etc by the pathogen type. Ordinary methods of removing the background nucleic acids in⁃clude physical separation, nuclease digestion and several rRNA depletion methods. The non-specific amplification methods of trace samples include random primer PCR method, SISPA method, anchored random primer PCR meth⁃od, RCA method, MDA method, NASBA method.
展开▼
