分子病理学在诊断学中的应用与研究

Research and Application of Molecular Pathology in Diagnostics

分子生物学技术的进步为分子诊断提供了新的思路,与传统的培养及表型鉴定相比,分子诊断技术大大缩短了诊断所需时间,同时提高了敏感性和特异性,且操作简便、易于重复。分子诊断方法大多在培养的基础上进行病原真菌分子鉴定,也有报道某些分子生物学技术可直接从液体培养瓶,甚至临床样本中进行病原检测。随着分子生物学理论和技术的不断发展,目前基于核酸和蛋白质的分子诊断技术已经开始逐步应用于临床,分子水平的诊断技术需要的时间短、特异性和敏感性高,在一定程度上克服了传统诊断方法的缺陷,可以实现对病原菌的早期、特异诊断,改善患者预后、提高患者生存率。核酸等温扩增技术能在一定温度下完成对DNA或RNA的扩增,与传统PCR方法相比,核酸等温扩增技术对仪器的要求极大地简化,恒温水浴即可完成反应,时间也大大地缩短,能满足实现操作简单、快速诊断的需求。环介导等温扩增技术针对基因序列的六个独立片段设计两对引物,实现对核酸片段的特异性扩增。

Advancesin molecular biology technology have provided new ideas for moleculardiagnosis.Compared with traditional culture and phenotypicidentification,molecular diagnostic technology has greatly shortened the timerequired for diagnosis,while improving sensitivity and specificity,and iseasy to operate and easy to repeat.Molecular diagnostic methods mostly performmolecular identification of pathogenic fungi on the basis of culture.It isalso reported that some molecular biology techniques can directly detectpathogens from liquid culture bottles and even clinical samples.With thecontinuous development of molecular biology theory and technology,moleculardiagnostic techniques based on nucleic acids and proteins have begun to begradually applied in clinical practice.The diagnostic techniques at themolecular level require short time,high specificity and sensitivity,andovercome to some extent.The defects of traditional diagnostic methods canachieve early and specific diagnosis of pathogenic bacteria,improve patientprognosis and improve patient survival rate.The nucleic acid isothermalamplification technology can complete the amplification of DNA or RNA at acertain temperature.Compared with the traditional PCR method,the nucleic acidisothermal amplification technology greatly simplifies the requirements of theinstrument,and the constant temperature water bath can complete the reaction,and the time is also greatly shortened;it can meet the needs of simple andfast diagnosis.Loop-mediated isothermal amplification techniques design twopairs of primers for six independent fragments of the gene sequence to achievespecific amplification of the nucleic acid fragments.