Loop-mediated amplification

Loop-mediated amplification (LAMP) Target DNA polymerase No... 

We demonstrated this principle here by using the isothermal NAAT known as Loop-Mediated Amplification (LAMP) in combination with a modification of the Enzymatic Luminometric Inorganic Pyrophosphate Detection Assay (ELIDA). This novel Bioluminescent Assay for Real-Time NAATs (BART-NAAT) is shown to be quantitative, rapid and cost-effective. 

Another method is loop-mediated isothermal DNA amplification (LAMP). This was carried out at a constant temperature of 65°C on the sample well of a PMMA chip. Subsequent CGE separation was carried out. The LAMP method has been applied for analysis of the PSA gene with a 23-fg/p.L template concentration [953,954],... 

Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 2000 28 E63. 

Nagamine K, Hase T, Notomi T. Accelerated reaction by loop-mediated isothermal amplification using loop primers. Mol Cell Probes 2(X)2 16 223-9. 

MIO. Mori, Y., Nagamine, K., Tomita, N., and Notomi, T., Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation. Biochem. Biophys. Res. Commun. 289, 150-154 (2001). 

Symbols NA, not applicable EP-PCR, endpoint PCR witii agarose gel electrophoresis LAMP, loop-mediated isodiermal amplification PCM, polycarbonate membrane CTAB, cetyltrimethylammonium bromide. 

Mori, Y., Notomi, T. (2009). Loop-mediated isothermal amplification (LAMP) a rapid, accurate, and cost-effective diagnostic method for infectious diseases. Journal of Infection and Chemotherapy, 15, 62-69. 

Tsuchiya, Y., Ogawa, M., Nakakita, Y, Nara, Y, Kaneda, H., Watari, J., et al. (2007). Identification of beer-spoilage microorganisms using the loop-mediated isothermal amplification method. Journal of the American Society of Brewing Chemists, 65, 77-80. 

Ahmed, M.U., Hasan, Q., Mosharraf Hossain, M. et al (2010) Meat species identification based on the loop mediated isothermal amplification and electrochemical DNA sensor. Food Control, 21 (5), 599-605. 

Nakamura, N., Fukuda, T., Nonen, S., Hashimoto, K., Azuma, J., Gemma, N. Simple and accurate determination of CYR2D6 gene copy number by a loop-mediated isothermal amplification method and an electrochemical DNA chip. Clin ChimActa 2010, 411, 568-573. 

EL-MATBOULI M and SOLIMAN H (2005), Rapid diagnosis of Tetracapsuloides bryo-salmonae, the causative agent of proliferative kidney disease (PKD) in salmonid fish by a novel DNA amplification method, loop-mediated isothermal amplification (LAMP), Parasitol Res, 96(5), 277-284. 

GUNiMALADEVi I, KONO T, LAPAiRA SE and SAKAI M (2005), A loop mediated isothermal amplification (LAMP) method for detection of infectious hematopoietic necrosis virus (IHNV) in rainbow trout (Oncorhynchus mykiss). Arch Virol, 150(5), 899-909. 

TTANO T, KAWAKAMi H, KONO T and SAKAI M (2006), Detection of fish nocardiosis by loop-mediated isothermal amplification, 7 App/ Microbiol, 100(6), 1381-1387. 

SAVAN R, KONO T,nAMiT and SAKAI M (2005), Loop-mediated isothermal amplification an emerging technology for detection of fish and shellfish pathogens, J Fish Dis, 28,573-581. 

WANG C-H, LIEN K-Y, WANG T-Y, CHEN T-Y and LEE G-B (2010), Integrated microfluidic loop-mediated-isothermal-amplification system for rapid diagnosis of aquaculture virus. 14 International Conference on Miniaturized Systems for Chemistry and Life Sciences, Groningen, The Netherlands, pp. 812-814. 

Moreover, a recent study also revealed that ROS generation led to the activation of caspase-2 during p-carotene-induced apoptosis in the human leukemic T cell line Molt 4. The apoptosis progressed by simultaneous activation of caspase-8 and caspase-9, and a cross talk between these initiator caspases was mediated by the pro-apoptotic protein Bid. Inhibition of caspases 2, 8, 9, and 3 independently suppressed the caspase cascade. The cleavage of the anti-apoptotic protein BclXL was found to be another important event during P-carotene-induced apoptosis, suggesting the presence of an extensive feedback amplification loop in P-carotene-induced apoptosis (Prasad et al., 2006). 

It is worth mentioning that in death receptor-mediated apoptosis, cells can be divided into two groups depending on the requirement for mitochondria to induce a complete apoptotic response. Type I cells do not require the mitochondrial pathway because the recruitment of procaspase-8 into the DISC complex is enough to fully activate caspase-8 which then activates effector caspases. However, Type II cells are characterized by an incomplete apoptotic response unless mitochondria are involved (Scaffidi et al., 1999). In this type of cell, efficient activation of effector caspases requires the mitochondrial amplification loop (Fig. 5). Caspase-8 cleaves cytosolic Bid, a BH3-only protein, which when cleaved to tBid is able to translocate to the mitochondria and trigger release of the proapoptotic factors cytochrome c and Smac/DIABLO (Li et al., 1998 Deng et al., 2002). The release of cytochrome c triggers apoptosome formation, subsequent caspase-9 activation, and finally the activation of effector caspases such as caspase-3. 

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