Fakruddin Md, Mannan Khanjada Shahnewaj Bin, Chowdhury Abhijit, Mazumdar Reaz Mohammad, Hossain Md Nur, Islam Sumaiya, Chowdhury Md Alimuddin
Industrial Microbiology Laboratory, Institute of Food Science and Technology, Bangladesh Council of Scientific and Industrial Research, Dhaka, Bangladesh.
J Pharm Bioallied Sci. 2013 Oct;5(4):245-52. doi: 10.4103/0975-7406.120066.
Nucleic acid amplification is a valuable molecular tool not only in basic research but also in application oriented fields, such as clinical medicine development, infectious diseases diagnosis, gene cloning and industrial quality control. A comperehensive review of the literature on the principles, applications, challenges and prospects of different alternative methods of polymerase chain reaction (PCR) was performed. PCR was the first nucleic acid amplification method. With the advancement of research, a no of alternative nucleic acid amplification methods has been developed such as loop mediated isothermal amplification, nucleic acid sequence based amplification, strand displacement amplification, multiple displacement amplification. Most of the alternative methods are isothermal obviating the need for thermal cyclers. Though principles of most of the alternate methods are relatively complex than that of PCR, they offer better applicability and sensitivity in cases where PCR has limitations. Most of the alternate methods still have to prove themselves through extensive validation studies and are not available in commercial form; they pose the potentiality to be used as replacements of PCR. Continuous research is going on in different parts of the world to make these methods viable technically and economically.
核酸扩增不仅是基础研究中的一种重要分子工具,在临床医学发展、传染病诊断、基因克隆和工业质量控制等应用导向领域也具有重要价值。本文对不同替代聚合酶链反应(PCR)方法的原理、应用、挑战及前景进行了全面的文献综述。PCR是第一种核酸扩增方法。随着研究的进展,已开发出许多替代核酸扩增方法,如环介导等温扩增、基于核酸序列的扩增、链置换扩增、多重置换扩增。大多数替代方法是等温的,无需热循环仪。尽管大多数替代方法的原理比PCR相对复杂,但在PCR存在局限性的情况下,它们具有更好的适用性和灵敏度。大多数替代方法仍需通过广泛的验证研究来证明自身价值,且尚无商业形式可用;它们具有替代PCR的潜力。世界各地都在持续开展研究,以使这些方法在技术和经济上可行。