Detection of gene point mutation in paraffin sections using in situ loop-mediated isothermal amplification

2007 ◽  
Vol 57 (9) ◽  
pp. 594-599 ◽  
Author(s):  
Satoshi Ikeda ◽  
Kazuhiko Takabe ◽  
Masaharu Inagaki ◽  
Naoya Funakoshi ◽  
Keiko Suzuki
Keyword(s):  
Point Mutation ◽  
Isothermal Amplification ◽  
Paraffin Sections ◽  
Loop Mediated Isothermal Amplification

scholarly journals In situ loop-mediated isothermal amplification (LAMP) for identification of Plasmodium species in wide-range thin blood smears

2018 ◽  
Vol 17 (1) ◽  
Author(s):  
Muneaki Hashimoto ◽  
Hirokazu Sakamoto ◽  
Yusuke Ido ◽  
Masato Tanaka ◽  
Shouki Yatsushiro ◽  
...  
Keyword(s):  
Plasmodium Species ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Blood Smears ◽  
Wide Range ◽  
Thin Blood Smears

scholarly journals Detection of the Benthic Dinoflagellates, Ostreopsis cf. ovata and Amphidinium massartii (Dinophyceae), Using Loop-Mediated Isothermal Amplification

2021 ◽  
Vol 9 (8) ◽  
pp. 885
Author(s):  
Eun Sun Lee ◽  
Jinik Hwang ◽  
Jun-Ho Hyung ◽  
Jaeyeon Park
Keyword(s):  
Lamp Assay ◽  
Isothermal Amplification ◽  
Detection Sensitivity ◽  
Loop Mediated Isothermal Amplification ◽  
Benthic Dinoflagellates ◽  
Field Samples ◽  
Its Gene ◽  
Buffer Composition ◽  
Species Specific

For the in situ and sensitive detection of benthic dinoflagellates, we have established an integrated loop-mediated isothermal amplification (LAMP) assay based on Ostreopsis cf. ovata and Amphidinium massartii. To detect the two species, a set of species-specific primers was constructed between the ITS gene and D1–D6 LSU gene, and the reaction temperature, time, and buffer composition were optimized to establish this method. In addition, the specificity of the LAMP primers was verified both in strains established in the laboratory and in field samples collected from the Jeju coastal waters, Korea. With the LAMP assay, the analysing time was within 45 to 60 min, which may be shorter than that with the conventional PCR. The detection sensitivity of the LAMP assay for O. cf. ovata or A. massartii was comparable to other molecular assays (PCR and quantitative PCR (qPCR)) and microscopy examination. The detection limit of LAMP was 0.1 cell of O. cf. ovata and 1 cell of A. massartii. The optimized LAMP assay was successfully applied to detect O. cf. ovata and A. massartii in field samples. Thus, this study provides an effective method for detecting target benthic dinoflagellate species, and could be further implemented to monitor phytoplankton in field surveys as an altenative.

scholarly journals Detection of Bacteria Carrying the stx2 Gene by In Situ Loop-Mediated Isothermal Amplification

2003 ◽  
Vol 69 (8) ◽  
pp. 5023-5028 ◽  
Author(s):  
Fumito Maruyama ◽  
Takehiko Kenzaka ◽  
Nobuyasu Yamaguchi ◽  
Katsuji Tani ◽  
Masao Nasu
Keyword(s):  
Cell Damage ◽  
Fluorescent Antibody ◽  
Dna Amplification ◽  
Isothermal Amplification ◽  
Specific Gene ◽  
Lamp Method ◽  
In Situ Pcr ◽  
Loop Mediated Isothermal Amplification ◽  
Antibody Labeling

ABSTRACT A new in situ DNA amplification technique for microscopic detection of bacteria carrying a specific gene is described. Loop-mediated isothermal amplification (LAMP) was used to detect stxA 2 in Escherichia coli O157:H7 cells. The mild permeabilization conditions and low isothermal temperature used in the in situ LAMP method caused less cell damage than in situ PCR. It allowed use of fluorescent antibody labeling in the bacterial mixture after the DNA amplification for identification of E. coli O157:H7 cells with an stxA 2 gene. Higher-contrast images were obtained with this method than with in situ PCR.

A loop-mediated, isothermal amplification-based method for visual detection of Vibrio parahaemolyticus within only 1 h, from shrimp sampling to results

2017 ◽  
Vol 9 (11) ◽  
pp. 1695-1701 ◽  
Author(s):  
Rui Wang ◽  
Xingning Xiao ◽  
Yuan Chen ◽  
Jun Wu ◽  
Wenjuan Qian ◽  
...  
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Visual Detection ◽  
Detection Method ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification

A quite simple, sensitive and visual detection method was developed forin situdetection ofV. parahaemolyticuswithin only 1 hour.

Ultraviolet-induced in situ gold nanoparticles for point-of-care testing of infectious diseases in loop-mediated isothermal amplification

Lab on a Chip ◽  
2021 ◽  
Vol 21 (4) ◽  
pp. 700-709
Author(s):  
Rajamanickam Sivakumar ◽  
Vu Phong Dinh ◽  
Nae Yoon Lee
Keyword(s):  
Gold Nanoparticles ◽  
Infectious Diseases ◽  
Enterococcus Faecium ◽  
Point Of Care ◽  
Isothermal Amplification ◽  
Point Of Care Testing ◽  
Loop Mediated Isothermal Amplification

The present study investigated ultraviolet-induced in situ gold nanoparticles (AuNPs) coupled with LAMP for the point-of-care testing (POCT) of two major infectious pathogens, namely, Coronavirus (COVID-19) and Enterococcus faecium (E. faecium spp.).

scholarly journals In-Situ Monitoring of Real-Time Loop-Mediated Isothermal Amplification with QCM: Detecting Listeria monocytogenes

Biosensors ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 308
Author(s):  
Sirirat Wachiralurpan ◽  
Isaratat Phung-On ◽  
Narong Chanlek ◽  
Supatra Areekit ◽  
Kosum Chansiri ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Real Time ◽  
Dna Sequences ◽  
Photoelectron Spectroscopy ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
In Situ Monitoring ◽  
Loop Mediated Isothermal Amplification ◽  
In Situ Detection

Functionalized DNA sequences are promising sensing elements to combine with transducers for bio-sensing specific target microbes. As an application example, this paper demonstrates in situ detection of loop-mediated isothermal amplification products by hybridizing them with thiolated-ssDNA covalently anchored on the electrodes of a quartz crystal microbalance (QCM). Such hybridization leads to a frequency signal, which is suitable for monitoring real-time LAMP amplification based on mass-sensing: it detects interactions between the complementary nucleobases of LAMP products in solution and the thiolated-ssDNA probe sequence on the gold surface. Target DNA LAMP products cause irreversible frequency shifts on the QCM surfaces during hybridization in the kHz range, which result from both changes in mass and charge on the electrode surface. In order to confirm the LAMP assay working in the QCM sensing system at elevated temperature, the sky blue of positive LAMP products solution was achieved by using the Hydroxy Naphthol Blue (HNB) and agarose gel electrophoresis. Since on-QCM sensing of DNA hybridization leads to irreversible sensor responses, this work shows characterization by X-ray photoelectron spectroscopy (XPS) core spectra of S2p, N1s, Mg1s, P2p and C1s. XPS results confirmed that indeed both DNA and by-products of LAMP attached to the surface. Listeria monocytogenes DNA served to study in-situ detection of amplified LAMP products on DNA-functionalized surfaces.

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