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

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

Loop-Mediated Isothermal Amplification in Microchambers on a Cell Microarray Chip for Identification of Plasmodium Species

2019 ◽  
Vol 105 (1) ◽  
pp. 69 ◽  
Author(s):  
Yusuke Ido ◽  
Muneaki Hashimoto ◽  
Shouki Yatsushiro ◽  
Masato Tanaka ◽  
Kazumichi Yokota ◽  
...  
Keyword(s):  
Plasmodium Species ◽  
Isothermal Amplification ◽  
Microarray Chip ◽  
Loop Mediated Isothermal Amplification ◽  
A Cell ◽  
Cell Microarray

Bench-scale experiments for the development of a unified loop-mediated isothermal amplification (LAMP) assay for the in vitro diagnosis of Leishmania species' promastigotes

2013 ◽  
Vol 142 (8) ◽  
pp. 1671-1677 ◽  
Author(s):  
M. KARANI ◽  
I. SOTIRIADOU ◽  
J. PLUTZER ◽  
P. KARANIS
Keyword(s):  
Lamp Assay ◽  
High Sensitivity ◽  
Leishmania Species ◽  
18S Rrna Gene ◽  
Isothermal Amplification ◽  
Rrna Gene ◽  
Bench Scale ◽  
Loop Mediated Isothermal Amplification ◽  
Wide Range

SUMMARYWe developed, in bench-scale experiments, a unified loop-mediated isothermal amplification (LAMP) assay for the detection of cutaneous, mucocutaneous and visceral leishmaniasis using DNA of cultivated promastigotes. Two primer sets for the LAMP assay were designed based on the 18S rRNA gene, and their sensitivity and specificity were tested and compared. Both of them were specific for Leishmania as the DNA of all ten Leishmania species tested was amplified, whereas the DNA of other parasites, including that of Trypanosoma, was not. The detection limit for primer set 1 ranged between 30 pg and 3·6 fg, depending on which Leishmania species tested. Primer set 2 showed high sensitivity, but was less sensitive than primer set 1. Our findings lead to the conclusion that the LAMP assay with primer set 1 is a promising and effective assay for the successful detection of a wide range of Leishmania infections using only a unified multiplex LAMP test.

scholarly journals Loop-Mediated Isothermal Amplification Assay for the Rapid Detection ofStaphylococcus aureus

2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
King Ting Lim ◽  
Cindy Shuan Ju Teh ◽  
Kwai Lin Thong
Keyword(s):  
Predictive Value ◽  
Food Poisoning ◽  
Lamp Assay ◽  
Rapid Identification ◽  
Negative Control ◽  
Isothermal Amplification ◽  
Promising Alternative ◽  
Loop Mediated Isothermal Amplification ◽  
Resource Limited ◽  
Wide Range

Staphylococcus aureus, including methicillin-resistantS. aureus(MRSA), is an important human pathogen that produces a variety of toxins and causes a wide range of infections, including soft-tissue infections, bacteremia, and staphylococcal food poisoning. A loop-mediated isothermal amplification (LAMP) assay targeting thearcCgene ofS. aureuswas developed and evaluated with 119S. aureusand 25 non-S. aureusstrains. The usefulness of the assay was compared with the PCR method that targetsspaandarcCgenes. The optimal temperature for the LAMP assay was 58.5°C with a detection limit of 2.5 ng/μL and 102 CFU/mL when compared to 12.5 ng/μL and 103 CFU/mL for PCR (spaandarcC). Both LAMP and PCR assays were 100% specific, 100% sensitive, 100% positive predictive value (PPV), and 100% negative predictive value (NPV). When tested on 30 spiked blood specimens (21 MRSA, eight non-S. aureusand one negative control), the performance of LAMP and PCR was comparable: 100% specific, 100% sensitive, 100% PPV, and 100% NPV. In conclusion, the LAMP assay was equally specific with a shorter detection time when compared to PCR in the identification ofS. aureus. The LAMP assay is a promising alternative method for the rapid identification ofS. aureusand could be used in resource-limited laboratories and fields.

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.

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