Loop-mediated Isothermal Amplification of DNA (LAMP): A New Diagnostic Tool Lights the World of Diagnosis of Animal and Human Pathogens: A Review

Abstract

Diagnosis is an important part in case of animal husbandry as treatment of a disease depends on it. Advancement in molecular biology has generated various sophisticated tools like Polymerase Chain Reaction (PCR), its versions along with pen-side diagnostic techniques. Every diagnostic test however has both advantages and disadvantages; PCR is not an exception to this statement. To ease the odds faced by PCR several non-PCR techniques which can amplify DNA at a constant temperature has become the need of hour, thus generating a variety of isothermal amplification techniques including Nucleic Acid Sequence-Based Amplification (NASBA) along with Self-Sustained Sequence Replication (3SR) and Strand Displacement Amplification (SDA) and Loop mediated isothermal amplification (LAMP) test. LAMP stands out to be a good and effective diagnostic test for empowering in developing countries as it does not require sophisticated equipments and skilled personnel and proves to be cost-effective. Performance of LAMP mainly relies on crafting of six primers (including 2 loop primers) ultimately accelerating the reaction. LAMP amplifies DNA in the process pyrophosphates are formed causing turbidity that facilitates visualisation in a more effective way than PCR. The Bst and Bsm polymerase are the required enzymes for LAMP that does not possess 5'-3' exonuclease activity. Results can be visualized by adding DNA binding dye, SYBR green. LAMP is more stable than PCR and real-time PCR. Non-involvement of template DNA preparation and ability to generate 10(9) copies of DNA are added benefits that make it more effective than NASBA or 3SR and SDA. Thus, it fetches researcher's interest in developing various versions of LAMP viz., its combination with lateral flow assay or micro LAMP and more recently lyophilized and electric (e) LAMP. Availability of ready to use LAMP kits has helped diagnosis of almost all pathogens. LAMP associated technologies however needs to be developed as a part of LAMP platform rather than developing them as separate entities. This review deals with all these salient features of this newly developed tool that has enlightened the world of diagnosis.

Arcobacter: an emerging food-borne zoonotic pathogen, its public health concerns and advances in diagnosis and control – a comprehensive review

“…For this purpose, further strengthening of the immunodiagnostics, molecular detection and characterization, genotyping and differentiation of Arcobacters is suggested by employing advances in PCR and its allied versions, LAMP, recombinant diagnostics, lateral flow assay, biochips, biosensors, microarrays, gene sequencing and phylogenetic analysis, genomic fingerprinting and nanotechnology-based diagnostics (Schmitt & Henderson 2005;Deb & Chakraborty 2012;Dhama et al 2012Dhama et al , 2014aNum & Useh 2013;Gilbert et al 2014;Ramees et al 2014d;Webb et al 2015Webb et al , 2016Whiteduck-L eveill ee et al 2016a). …”

Loop-mediated isothermal amplification (LAMP): a versatile technique for detection of micro-organisms

“…This is due to the extremely high efficiency of the LAMP reaction. The LAMP product is so stable that it is not degraded easily and thus unintended carryover contamination can occurs (Bi et al 2012;Dhama et al 2014;Fischbach et al 2015). In case aforesaid contamination occurs, it is suggested to perform LAMP in a laminar flow hood and use separate pipettes and filtered tips.…”

Zika virus – emergence, evolution, pathology, diagnosis, and control: current global scenario and future perspectives – a comprehensive review

“…Till now, no commercial diagnostic tests exist for diagnosing this viral infection. Advances in diagnostic techniques need to be applied for Zika virus detection viz., multiplex PCR, LAMP, recombinant diagnostics, gene sequencing and phylogenetic analysis, biosensors, biochips, microarrays, and nanotechnologybased diagnostics (Jain 2007;Kawadkar et al 2011;Dhama, Karthik, et al 2014;Ratcliff et al 2007;Bergquist 2011;Van den Hurk and Evoy 2015). Apart from these newer tools for extensive epidemiological and surveillance studies, alternatives need to be explored for tracking and monitoring Zika virus infections and spread, such as geographical information system and establishing networking programs to counter this important pathogen.…”