reproduce this study has good specificity and sensitivity,
reproduce inside the host’s cells and is one of the most virulent foodborne pathogens. The LAMP method of Listeria monocytogenes established in this study has good specificity and sensitivity, which can be applied to the rapid detection of Listeria monocytogenes. Acetic acid bacteria can contaminate food. Acetobacter aceti is a subgroup of acetic acid bacteria. It is usually isolated from yogurt. A. aceti should be timely and effectively detected to prevent yogurt contamination. In a study which performed to detect acetic acid bacteria in yogurt. LAMP, PCR, and real-time PCR were applied and compared for detecting A. aceti from pure culture and artificially contaminated yogurt samples assume that in pure culture, LAMP showed the highest detection sensitivity with 10-2 CFU/mL. For yogurt samples, the sensitivity limit of LAMP was 102 CFU/mL, which was lower than that of real-time PCR (101 CFU/mL). The results indicated that these methods could be quickly and efficiently applied to detect A. aceti. As LAMP technology has low cost and high detection efficiency, it can potentially be applied for detecting A. aceti in production and quality control programs of yogurt. 15.Environmental testing: Cryptosporidiosis can cause disease. It has major public health concern. Genotypically and phenotypically diverse Cryptosporidium species are responsible for these disease. The impact on water and food in the epidemiology of this disease is well determined. A loop-mediated isothermal amplification (LAMP) procedure for the detection of Cryptosporidium in environmental and fecal samples is developed and evaluated. LAMP can be a potential diagnostic tool which can examine Cryptosporidium spp. in samples for clinical laboratories and water industries due to its simplicity, specificity, and cost-effectiveness. Rapid detection of airborne pathogen are very important for disease prevention and public safety. It also indicate the biological air pollution. In a study, a microfluidic system is established that could capture and enrich airborne pathogens in high-throughput LAMP analysis. Five species of bacteria show good stability and specificity for this method. Approximate detection limit down to 24 cells per reaction is achieved for Staphylococcus aureus,without the process of DNA purification. 16.Plant species identification in food: An application of LAMP methods which can detect closely related plant species in food or feed matrices has been established, In a study, LAMP-based methods for plant species identification with respect to method parameters such as R2, LOD, and LOQ is evaluated. An existing (real-time) PCR method (for the detection of spices) is used for comparison in this study. It could be shown that the developed LAMP methods have potential as alternative strategies to PCR in DNA-based analysis. 17.SNP typing: LAMP method is highly specific. So it can be used in single nucleotide polymorphism (SNP) detection. Only the target gene will be amplified from gene samples containing homologous nucleotide sequences when using LAMP-based SNPs typing. LAMP method can distinguish a single nucleotide difference at each cycling step of the DNA replication because of its reaction characteristics, through both “sense and anti-sense strand” reactions, LAMP can easily detected SNP in a single step.. LAMP method is very simple and rapid. For that, SNPs typing is detected within 30 minutes.4 primers designed to recognize 6 distinct regions, so they amplify only the target gene is strictly and specifically .LAMP can detect SNP even in coexistence with its h homologous gene. The reaction is so specific so that single nucleotide difference is strictly discriminated. The SNPs typing can be performed by in a single step. In figure (16) basic principal of SNP typing with LAMP is illustrated. Figure (16): LAMP based SNP typing 18.Application in agricultural sector: It is very necessary to develop a rapid and precise method that can diagnose rice viruses in host plants and in vector insects. It is very important to control rice virus diseases. A reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay, is developed. It detects nine viruses, including eight RNA viruses and one DNA virus, in infected rice plants and the viruliferous vector insects. The sensitivities of the assays are higher or similar to those of one-step RT-PCR. Rapid. RNA extraction is done by using RNA extraction kit .It combine with RT-LAMP assay. Nine viruses are detected within 2 h from infected rice plants and the viruliferous insects. It is inexpensive and avoid delicate equipment. RT-LAMP method for rice viruses can significant for the epidemiology and molecular pathology of rice viruses study. The cry2Ab and cry3A are exogenous genes which are most important for express insect resistant protein in GM Plant .This two genes come from Bacillus thuringenesis cry protein family. Cry is widely used in genetically-modified (GM) crops. It is very important to develop an effective techniques for the rapid detection of these genes ingenetically-modified organisms (GMOs). A rapid and visual loop-mediated isothermal amplification (LAMP) method is developed to detect the cry2Ab and cry3A genes. The LAMP assay can be finished within 60 min at an isothermal condition of a constant temperature 60-65 °C. The derived LAMP products can be obtained by a real-time turbidimeter via monitoring the white turbidity or directly observed by the naked eye through adding SYBR Green I dye. In addition, this method does not need specific expensive instruments or facilities, which can provide a simpler and quicker approach to detecting the cry2Ab and cry3A genes in GM crops. One significant fact is this method can run on-site, large-scale test purposes in the field. Bt brinjal, Bt corn, Bt sugarcane can be used efficiently in this method. Verticillium dahliae is a fungal plant pathogen. It causes verticillium wilt in many plant species, causing leaves to curl and discolor. It may cause death in some plants. All over the world there are 400 plant species that can be affected by Verticillium . A loop-mediated isothermal amplification (LAMP) assay has been developed and evaluated for the rapid and sensitive detection of Verticillium dahliae Kleb. LAMP primers design based on a known RAPD marker, No DNA purification step is needed. This assay is applied on V. dahliae grown on medium and from soil samples. In a study ,the detection ratios of LAMP protocols were better (26 positive samples out of 32 agricultural soils analysed, respectively) than that obtained for nested-PCR method (22 positive results). Dacus ciliatus Loew (Diptera: Tephritidae), is a cucumber fruit fly is one of the most destructive agricultural pests which attack mainly fruits of Cucurbitaceae. This pest is widespread and highly invasive; thus,. So It is a high priority for pest detection and quarantine programs.Although ,using morphological keys ,cucumber fruit fly adults can be identified and distinguished from the other species. But it is often difficult to distinguish this species from the other tephritids that share host plants by using material from other stages of development. So a quick and efficient alternative species diagnostic tool would be valuable for this. (PCR-LAMP) is a combined method of LAMP with PCR , present for the rapid detection and discrimination of cucumber fruit fly DNA from some other common tephritid species attacking Cucurbitaceae, using material from different stages of development. The method is species-specific and sensitive and provide a rapid diagnostic tool to detect D. ciliaus even it can be run by non-experts. Pythium species can cause diseases in economically important plant. It have been frequently found to cause diseases in soilless greenhouse systems. Pythium can contaminate irrigation water supplies. It also spread rapidly in hydroponically grown crops. Early infection by Pythium species causes yield losses in many vegetables and ornamental crops. Pythium myriotylum Drechsler is a causal agent of root rot in economically important crops including peanuts, tomato, rye, wheat, oats, cucumber, soya bean, sorghum, tobacco, cabbage and maize. A loop-mediated isothermal amplification (LAMP) reaction for the detection of Pythium myriotylum is present. The primer set targeting the ITS sequence of P. myriotylum .which work most efficiently at 60°C and allowed Detection of DNA can be performed within 30 min by fluorescence monitoring using a real-time PCR instrument. LAMP can detect P. myriotylum in hydroponic solution samples, and the results are very similar with conventional plating method in almost all cases. Grape downy mildew, caused by Plasmopara viticola, can cause Grape downy mildew. It is one of the most important diseases of grapes worldwide. The pathogen can infect all green parts of the vine when the environment is warm and wet in the growing season. After years of extremely favorable environmental conditions, 80% of yield reduction may be observed. Chemical control is the effective solution for grape downy mildew. Although, these chemicals often alert use in absence of infection. So the extreme use of chemicals may occur environment pollution and can also reduce grape quality. Rapid and sensitive detection techniques for monitoring the presence and severity of infection would help growers to apply chemicals with more accuracy and at a higher efficiency, without the risk of crop loss. A LAMP-based assay is developed for the specific detection of P. viticola in infected leaves and the airborne sporangia collected from a spore trap. The use of the visual colorimetric indicator hydroxy naphthol blue for the in-tube detection of DNA amplification as well as simplified methods for the preparation of amplifiable target DNA. It may widely improve disease management in the grape industry by allowing the pathogen to be monitored and making analyses easier and more cost effective. The P. viticola ITS sequence (DQ665668.1) was chosen as the target region for the LAMP primers. For purified DNA, LAMP was at least 100 fold more sensitive than conventional PCR in this process. Fusarium graminearum, is a plant pathogen. It fusarium head blight on wheat and barley. The pathogen is responsible for billions of dollars of economic losses in worldwide every year. (LAMP) procedure use to detect genomic DNA of Fusarium graminearum in contaminated wheat seeds. There is no need to use liquid nitrogen for crushing germinated seedlings. The method do not take not more than one hour to prepare high quality DNA. In combination with the LAMP assay it is a fast and cost-effective. So it may be a alternative to traditional diagnostic methods for the early detection of toxigenic fusaria in cereals. Candidatus Liberibacter solanacearum (Lso) bacteria cause potato zebra chip (ZC) disease in potato and make million dollars loss in potato industry. PCR can be used for identification of Candidatus Liberibacter. But that requires agarose gel electrophoresis for resolution, on the other hand the positive LAMP results can be visualized directly as a precipitate within the reaction tubes. The 16S rDNA gene of “Ca. Liberibacter solanacearum” use as the target for the design of the six LAMP primers. The LAMP technique is a reliable, rapid, and cost-effective method. It can detects the “Ca. Liberibacter solanacearum” pathogen in the potato/tomato psyllid, Bactericera cockerelli, and in field-grown potato plants and tubers in high specificity. 19.Herbal medicine identification: Identification of medicinal plant ingredients is important for drug industry. It also ensure safe use and for the regulation of herbal drug supply chain. Loop-mediated isothermal amplification (LAMP) can identify herbal medicine species very efficiently. 23.Future aspects of LAMP: LAMP may spread in the field level of molecular biology and under privileged laboratories all over the world near future because of its cost effectiveness, high specificity, rapidity, without problem of inhibitor and successful amplification of DNA from a few copies. It will make forensic DNA test much easier and inexpensive. LAMP testing kits on microchip may use in future in both developed and developing countries. 24.Discussion: In 1953, the discovery of DNA was a breakthrough in the field of biological science. After that various technique related with detection and amplification of nucleic acid were invented. In 1985 PCR has invented as a revolutionary technique in molecular biology which is a rapid and simple technique that can produce large number copies of DNA copies from minute amount of DNA. . Loop mediated isothermal amplification (LAMP) is a versatile technique in molecular biology that can amplify DNA an isothermal condition. For some significant reason LAMP become a superior technique than PCR in various field of molecular biology. In LAMP, the amplification efficiency is extremely high withstand it is noticeably cost effective. LAMP do not need any thermal cycler and performed in heat block or water bath in a single tube with only two steps. Post purification step and highly pure DNA do not requires for LAMP method. These make LAMP a very impressive and potential method for developing and under developed country, because of costly instrument and highly trained personnel are tough to bear in their resource limited laboratory, specifically laboratory use for disease diagnosis purpose. LAMP can be performed on site of agriculture field for identifying specific plant pathogen and in water reservoir for detecting the pollution of water by biological substances very rapidly. So it make LAMP farmer friendly and according to spirit of age technique. LAMP uses four primers which can detect six region of gene. It makes LAMP extremely specific and highly sensitive even more than PCR in several cases. Another fact is about the analysis of LAMP reaction product. LAMP reaction produced amplicons possible to detect without use of gel electrophoresis, only with naked eye and turbidity. LAMP rapidity and high specificity make it remarkable for infectious pathogen detection in human. It work for both DNA and RNA and do not bother about matrix inhibitor. So LAMP’s accuracy of detection is more than conventional PCR and also give false positive result less than PCR. Therefore, LAMP method paves a new way to diagnose pathogenic microorganism and parasite in clinical laboratories. It can amplify from DNA a few copies of DNA. It is possible to early detection of pathogen and parasite with both rapidly and inexpensively by LAMP. So it can be a life savior techniques for human. Genetically modified organism (GMO) is a most popular word in science world nowadays. LAMP is proved as a very efficient technique to detect inserted gene in field level plants in a large scale. It is very easy technique to determine horizontal foreign gene transfer, transferred inserted gene in wild type. LAMP also have some noticeable drawbacks. This method do not work for unknown sequence. Although, complication related with primer designing is reducing because of several primer design software and tools are available and enriching day by day. Consequently, it will be speculated that, in near future LAMP method will become more precise and expanded in molecular biology and clinical laboratory and the application also increase day by day. 25.Conclusion: LAMP method is a blessings for developing countries at molecular biology sector. Even it uses very successfully in developed countries because of disease diagnosis proficiency, agricultural application and cost effectiveness. It is obligatory to execute LAMP method is resource limited laboratories.