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.
Full-textDOI: · Available from: Kuldeep Dhama, Jan 23, 2014
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ABSTRACT: Brucella abortus, one of the major pathogen causing abortions in cattle worldwide and a zoonotic agent, need to be detected earlier in order to prevent its spread among animals. The present study aimed at to know the prevalence of B. abortus in cattle population of three states (Uttar Pradesh, Uttarakhand and Tamil Nadu) of India by serological [Rose Bengal Plate Test (RBPT) and Serum Tube Agglutination Test (STAT)] and molecular (Polymerase Chain Reaction) detection in sera samples and whole blood (n=370), respectively. Out of a total of 370 sera samples, 61 (16.49%) were positive by RBPT and 59 (15.94%) by STAT. Screening of the whole blood samples by genus specific bcsp31 gene based PCR as well as species specific IS711 gene based PCR revealed that 56 (15.13%) samples were positive for brucellosis. None of the serologically negative sample showed positivity by PCR; however few positive samples were tested negative by PCR. Sensitivity and specificity of PCR compared with RBPT was 100% and 92.4%, while with STAT these were 100% and 95.16%, respectively. Results are promising that whole blood can be used for studying the molecular epidemiology of B. abortus in cattle, and particularly detecting the active phase of infection, and PCR can be well adopted as a valuable test for mass screening of animals for this purpose. The present study adds to the prevalence data available regarding to B. abortus infection in cattle population, and highlights the usefulness and advantages of molecular tool of PCR over serological tests.Asian Journal of Animal and Veterinary Advances 04/2014; 9(4):262-269. DOI:10.3923/ajava.2014.262.269 · 0.87 Impact Factor
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ABSTRACT: In this review, perspective roles and applications of molecular tools and techniques boosting production potentials and protection from economically important diseases are discussed. Importance of advent in genetic engineering, designer products, disease resistant plants and animals, advanced diagnostics, networking programmes, effective and safer vaccines as well as novel and emerging therapies are highlights of the review. The production of designer foods along with progress in ‘Omics’ as well as analytical fields contributed immensely for safeguarding health of livestock. For plunging losses caused to plant and animal population from pathogens, numerous methods are being adopted by biologists for engineering resistant breeds of animals and plants during last two decades. Advancements have been made in development of innovative detection methods (including modification of polymerase chain reaction) which have contributed in clinical decision making procedures and to determine the molecular epidemiology. Several new generation vaccines viz. DNA vaccines, reverse genetics vaccines, vector based vaccines, protein or peptide-based vaccines, gene-deleted vaccines and chimeric vaccines etc. have been developed or are under developmental phase for combating a wide array of diseases. Mapping for location of herds and flocks, enrichment of knowledge of epidemiologists and diagnosticians, clinicians along with researchers has become possible with the advancement of geographical information system (GIS). In addition bacteriophage, virophage, cytokine, probiotics as well as herbal, nutritional, immunomodulation therapy and cow (panchgavya) therapy have proved to be beneficial. Among them many are cost-effective and cause only minimal adverse reactions. This review gives a brief presentation of all the important innovative technologies boosting production and safeguarding health of humans and animals.Research Opinions in Animal and Veterinary Sciences 04/2014; 4(7):353-370.
- Advances in Animal and Veterinary Sciences 01/2014;