Nucleic acid amplification: Alternative methods of polymerase chain reaction

Abstract

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.

Future developments in biosensors for field-ready Zika virus diagnostics

“…Fortunately, over the past 30 years isothermal amplification techniques with typical amplification times of less than 1 h have been thoroughly described for a variety of DNA and RNA targets. Popular forms of isothermal NAATs include nucleic acid sequence-based amplification (NASBA), loop-mediated isothermal amplification (LAMP), strand invasion based amplification (SIBA), strand displacement amplification (SDA), helicase-dependent amplification (HAD), recombinase polymerase amplification (RPA) and others [38].

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Experimental metagenomics and ribosomal profiling of the human skin microbiome

“…This may be performed prior to or as part of library preparation. [77,78] Several amplification methods are available, [79] such as multiple displacement amplification [80] or limited-cycle PCR which is used in the popular Illumina NexteraXT Kit (Illumina®, San Diego,CA). [81] Regarding the choice of the sequencing platform, Illumina has become the de facto standard for metagenomic sequencing mainly because of its high throughput.…”

The Significance of Vitamin D Receptor Gene Polymorphisms for Susceptibility to Hepatocellular Carcinoma in Subjects Infected with Hepatitis C Virus

“…Genotyping was performed using a commercial PCR-SSP Sequence-Specific Primer (Olerup SSP; One Lambda Inc., Canoga Park, California, USA). VDR genotypes were identified using PCR amplification followed by restriction fragment length polymorphisms (PCR-RFLP) assay [35]. Two fragments of VDR gene were amplified.…”