Metode Loop-Mediated Isothermal Amplification (LAMP) dan Aplikasinya untuk Deteksi Penyakit Ikan

Polymerase chain reaction (PCR) is a method that amplifies DNA which have been widely used in molecular biology technique. Based on the PCR, many methods have been developed on isothermal condition and the useful one is loop-mediated isothermal amplification of DNA (LAMP). LAMP reaction employs a Bst DNA polymerase and a set of four specific primers that recognizes a total of six distinct sequences of the target DNA and produces amount of different size of DNA. Many advantages have been achieved in LAMP such as the simple equipment for reaction and observation, short time, highly specific and sensitive procedure. LAMP has been used as a tools for detection many pathogens for human, animals and plants. Some fish pathogens as parasites, bacteria and viruses have been detected by LAMP. The principles and application of LAMP method are discussed in this paper.

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Loop-Mediated Isothermal Amplification Method for Rapid Detection of Shigella dysenteriae

Applied Mechanics and Materials ◽

10.4028/www.scientific.net/amm.140.369 ◽

2011 ◽

Vol 140 ◽

pp. 369-373 ◽

Cited By ~ 2

Author(s):

Qing Ping Zhong ◽

Li Wang ◽

Bin Wang ◽

Hong Yuan Chen

Keyword(s):

Isothermal Condition ◽

Lamp Assay ◽

High Specificity ◽

Isothermal Amplification ◽

Shigella Dysenteriae ◽

Lamp Method ◽

Loop Mediated Isothermal Amplification ◽

Amplification Method ◽

Target Dna ◽

Pcr Method

The study was aimed to develop a loop-mediated isothermal amplification (LAMP) method which amplifies DNA with high specificity and rapidity for the detection of Shigella dysenteriae. A set of four primers was designed for recognizing six distinct sequences on the target ipaH of S. dysenteriae. By the method, the target DNA was amplified within 1h under isothermal condition at 65 °C. The sensitivities of the LAMP for detecting pure culture and genomic DNA were 1.04 CFU/ml and 1.06 fg/μl, while the sensitivities of PCR method were 1.04×102 CFU/ml and 1.06 pg/μl. Furthermore, the LAMP assay was examined for its ability to detect S. dysenteriae in artificially contaminated lettuce sample, the detection limits of this LAMP assay and the PCR method were 4.60 CFU/g and 4.60×102 CFU/g, respectively.

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Loop-Mediated Isothermal Amplification for Detection ofStaphylococcus aureusin Dairy Cow Suffering from Mastitis

Journal of Biomedicine and Biotechnology ◽

10.1155/2012/435982 ◽

2012 ◽

Vol 2012 ◽

pp. 1-5 ◽

Cited By ~ 9

Author(s):

Zhang Tie ◽

Wang Chunguang ◽

Wei Xiaoyuan ◽

Zhao Xinghua ◽

Zhong Xiuhui

Keyword(s):

Staphylococcus Aureus ◽

High Sensitivity ◽

High Specificity ◽

Isothermal Amplification ◽

Lamp Method ◽

Specific Primers ◽

Loop Mediated Isothermal Amplification ◽

Reaction Tube ◽

Specificity And Sensitivity ◽

Detectable Concentration

To develop a rapid detection method ofStaphylococcus aureususing loop-mediated isothermal amplification (LAMP), four specific primers were designed according to six distinct sequences of thenucgene. In addition, the specificity and sensitivity of LAMP were verified and compared with those of PCR. Results showed that the LAMP reaction was completed within 45 min at 62.5°C, and ladder bands were appeared in LAMP products analyzed by gel electrophoresis. After adding 1x SYBR Green l, the positive reaction tube showed green color and the negative reaction tube remained orange, indicating that the LAMP has high specificity. The minimal detectable concentration of LAMP was1×102 CFU/mL and that of PCR was1×104 CFU/mL, indicating that the LAMP was 100 times more sensitive than the PCR. The LAMP method for detection ofStaphylococcus aureushas many advantages, such as simple operation, high sensitivity, high specificity, and rapid analysis. Therefore, this method is more suitable for the rapid on-site detection ofStaphylococcus aureus.

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Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of viable but non-culturable Vibrio cholerae O1

Canadian Journal of Microbiology ◽

10.1139/cjm-2021-0142 ◽

2021 ◽

Author(s):

Parastoo Chamanrokh ◽

Rita R. Colwell ◽

Anwar Huq

Keyword(s):

Vibrio Cholerae ◽

Culture Media ◽

Lamp Assay ◽

Environmental Water ◽

Isothermal Amplification ◽

Lamp Method ◽

Waterborne Pathogen ◽

Loop Mediated Isothermal Amplification ◽

Added Benefit ◽

Polymerase Chain

Vibrio cholerae, an important waterborne pathogen, is a rod-shaped bacterium that naturally exists in aquatic environments. When conditions are unfavorable for growth, the bacterium can undergo morphological and physiological changes to assume a coccoid morphology. This stage in its life cycle is referred to as viable but non-culturable (VBNC) since VBNC cells do not grow on conventional bacteriological culture media. The current study compared polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) to detect and identify VBNC V. cholerae. Because it is difficult to detect and identify VBNC V. cholerae, the results of the current study are useful in showing LAMP to be more sensitive and rapid than PCR in detecting and identifying non-culturable, coccoid forms of V. cholerae. Furthermore, the LAMP method is effective in detecting and identifying very low numbers of coccoid VBNC V. cholerae in environmental water samples, with the added benefit of being inexpensive to perform.

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The Sensitivity and Specificity of Loop-Mediated Isothermal Amplification and PCR Methods in Detection of Foodborne Mi-croorganisms: A Systematic Review and Meta-Analysis

Iranian Journal of Public Health ◽

10.18502/ijph.v50i11.7571 ◽

2021 ◽

Author(s):

Yasaman Sadeghi ◽

Pegah Kananizadeh ◽

Solmaz Ohadian Moghadam ◽

Ahad Alizadeh ◽

Mohammad Reza Pourmand ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Likelihood Ratio ◽

Gold Standard ◽

Meta Analysis ◽

Isothermal Amplification ◽

Lamp Method ◽

Chain Reaction ◽

Cultivation Method ◽

Loop Mediated Isothermal Amplification ◽

Polymerase Chain

Background: The loop-mediated isothermal amplification (LAMP) method is frequently used for identifying many microorganisms. The present review aimed to evaluate the sensitivity and specificity of LAMP method for detection of food-borne bacteria and to compare these features with those of polymerase chain reaction (PCR), as an alternative molecular diagnostic procedure, and with cultivation method, as the gold standard method. Methods: The literature was searched in electronic databases (PubMed, Scopus, Web of Science, and EMBASE) for recruiting publications within Jan 2000 to Jul 2021. We used the combinations of keywords including foodborne disease, LAMP, PCR, Loop-mediated isothermal amplification, and polymerase chain reaction. Meta-analysis was used to adjust the correlation and heterogeneity between the studies. The efficiency of the methods was presented by negative likelihood ratio, positive likelihood ratio, sensitivity, specificity, and odds ratio using forest plots. A P-value less than 0.05 was considered as statistical significance cut off. The confidence intervals were presented at the 95% interval. Results: Overall, 23 relevant studies were analyzed. The sensitivities of LAMP and PCR methods were estimated to be 96.6% (95% CI: 95.0-97.7) and 95.6% (95%CI: 91.5-97.8), respectively. The specificities of LAMP and PCR were also estimated to be 97.6% (95%CI: 92.6-99.3) and 98.7% (95%CI: 96.5-99.5), respectively. Conclusion: The specificities of LAMP and PCR assays were determined by comparing their results with cultivation method as the gold standard. Overall, the specificity of both PCR and LAMP methods was low for detection of fastidious bacteria. Nevertheless, LAMP and PCR methods have acceptable specificities and sensitivities, and their application in clinical practice necessitates more studies.

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Loop-mediated Isothermal Amplification for Rapid Detection of Mating Types of Villosiclava virens

Plant Disease ◽

10.1094/pdis-09-21-1943-re ◽

2021 ◽

Author(s):

Xiayan Pan ◽

Xiao Wang ◽

Junjie Yu ◽

Mina Yu ◽

Huijuan Cao ◽

...

Keyword(s):

Mating Type ◽

Genomic Dna ◽

High Efficiency ◽

Lamp Assay ◽

Isothermal Amplification ◽

Loop Mediated Isothermal Amplification ◽

Villosiclava Virens ◽

False Smut ◽

Mating Type Genes ◽

Polymerase Chain

Rice false smut (RFS), caused by Villosiclava virens, is an important fungal disease in panicle of rice. V. virens is a heterothallic ascomycete that controlled by two opposite idiomorphs, MAT1-1 and MAT1-2. Previous study showed sexual reproduction of V. virens plays an important role in the epidemic of RFS. In this study, we have developed a loop-mediated isothermal amplification (LAMP) assay to detect mating type of V. virens easily and rapidly by using specific primers designed on the mating type genes MAT1-1-2 and MAT1-2-1, respectively. The LAMP assay required only a water/dry bath and could recognize the mating type of V. virens in just 45 min. The LAMP assay was so sensitive that could detect small amounts of V. virens genomic DNA (as low as 2.0 pg of MAT1-1, and 200.0 pg of MAT1-2), which was 10-fold more sensitive than polymerase chain reaction (PCR). In addition, the application of mating type using LAMP assay was demonstrated feasibly by assessing the genomic DNA of V. virens isolated from rice fields. The high efficiency and specificity of this LAMP assay suggested it can be used as a rapid testing tool in mating type recognition of V. virens isolates in the field.

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