scholarly journals Real-time loop-mediated isothermal amplification assay for rapid detection of Fusarium circinatum

BioTechniques ◽  
2020 ◽  
Vol 69 (1) ◽  
pp. 11-17 ◽  
Author(s):  
Dagmar Stehlíková ◽  
Nicola Luchi ◽  
Chiara Aglietti ◽  
Alessia Lucia Pepori ◽  
Julio Javier Diez ◽  
...  
Keyword(s):  
High Efficiency ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Pitch Canker ◽  
Portable Instrument ◽  
Fusarium Circinatum ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Assay ◽  
Time Loop ◽  
Host Tissues

Fusarium circinatum is the causal agent of pitch canker, a lethal disease of pine and other conifers. Since F. circinatum is a quarantine organism, its timely detection could efficiently prevent its introduction into new areas or facilitate spread management in already infected sites. In this study, we developed a sequence-specific probe loop-mediated isothermal amplification (LAMP) assay for F. circinatum using a field-deployable portable instrument. The assay was able to recognize the pathogen in host tissues in just 30 min, and the sensitivity of the assay made it possible to detect even small amounts of F. circinatum DNA (as low as 0.5 pg/μl). The high efficiency of this method suggests its use as a standard diagnostic tool during phytosanitary controls.

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

Plant Disease ◽  
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.

scholarly journals Development of a Genomics-Based LAMP (Loop-Mediated Isothermal Amplification) Assay for Detection of Pseudomonas fuscovaginae from Rice

Plant Disease ◽  
2014 ◽  
Vol 98 (7) ◽  
pp. 909-915 ◽  
Author(s):  
Gavin J. Ash ◽  
Jillian M. Lang ◽  
Lindsay R. Triplett ◽  
Benjamin J. Stodart ◽  
Valérie Verdier ◽  
...  
Keyword(s):  
Draft Genome ◽  
Lamp Assay ◽  
Brown Rot ◽  
Isothermal Amplification ◽  
Field Surveys ◽  
Next Generation Sequencing Technology ◽  
Loop Mediated Isothermal Amplification ◽  
Pure Cultures ◽  
Amplification Assay ◽  
Bacterial Plant Pathogen

The vast amount of data available through next-generation sequencing technology is facilitating the design of diagnostic marker systems. This study reports the use of draft genome sequences from the bacterial plant pathogen Pseudomonas fuscovaginae, the cause of sheath brown rot of rice, to describe the genetic diversity within a worldwide collection of strains representing the species. Based on a comparative analysis with the draft sequences, primers for a loop-mediated isothermal amplification (LAMP) assay were developed to identify P. fuscovaginae. The assay reported here reliably differentiated strains of P. fuscovaginae isolated from rice from a range of other bacteria that are commonly isolated from rice and other plants using a primer combination designated Pf8. The LAMP assay identified P. fuscovaginae purified DNA, live or heat-killed cells from pure cultures, and detected the bacterium in extracts or exudates from infected host plant material. The P. fuscovaginae LAMP assay is a suitable diagnostic tool for the glasshouse and laboratory and could be further developed for in-field surveys.

A sequence-specific plasmonic loop-mediated isothermal amplification assay with orthogonal color readouts enabled by CRISPR Cas12a

2020 ◽  
Vol 56 (24) ◽  
pp. 3536-3538 ◽  
Author(s):  
Rongxing Zhou ◽  
Yongya Li ◽  
Tianyu Dong ◽  
Yanan Tang ◽  
Feng Li
Keyword(s):  
Lamp Assay ◽  
Isothermal Amplification ◽  
Loop Mediated Isothermal Amplification ◽  
Complementary Color ◽  
Amplification Assay

CRISPR Cas12a enables a sequence-specific plasmonic LAMP assay with dual complementary color readouts.

Development of a Loop-Mediated Isothermal Amplification Assay for Rapid, Sensitive Detection of Campylobacter jejuni in Cattle Farm Samples

2014 ◽  
Vol 77 (9) ◽  
pp. 1593-1598 ◽  
Author(s):  
HEE-JIN DONG ◽  
AE-RI CHO ◽  
TAE-WOOK HAHN ◽  
SEONGBEOM CHO
Keyword(s):  
Campylobacter Jejuni ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Sensitive Detection ◽  
Cattle Farm ◽  
Loop Mediated Isothermal Amplification ◽  
Amplification Assay ◽  
Pcr Assays ◽  
Template Dna ◽  
Commercial Kit

Campylobacter jejuni is a leading cause of bacterial foodborne disease worldwide. The detection of this organism in cattle and their environment is important for the control of C. jejuni transmission and the prevention of campylobacteriosis. Here, we describe the development of a rapid and sensitive method for the detection of C. jejuni in naturally contaminated cattle farm samples, based on real-time loop-mediated isothermal amplification (LAMP) of the hipO gene. The LAMP assay was specific (100%inclusivity and exclusivity for 84 C. jejuni and 41 non–C. jejuni strains, respectively), sensitive (detection limit of 100 fg/μl), and quantifiable (R2 = 0.9133). The sensitivity of the LAMP assay was then evaluated for its application to the naturally contaminated cattle farm samples. C. jejuni strains were isolated from 51 (20.7%) of 246 cattle farm samples, and the presence of the hipO gene was tested using the LAMP assay. Amplification of the hipO gene by LAMP within 30 min (mean =10.8 min) in all C. jejuni isolates (n = 51) demonstrated its rapidity and accuracy. Next, template DNA was prepared from a total of 186 enrichment broth cultures of cattle farm samples either by boiling or using a commercial kit, and the sensitivity of detection of C. jejuni was compared between the LAMP and PCR assays. In DNA samples prepared by boiling, the higher sensitivity of the LAMP assay (84.4%) compared with the PCR assay (35.5%) indicates that it is less susceptible to the existence of inhibitors in sample material. In DNA samples prepared using a commercial kit, both the LAMP and PCR assays showed 100% sensitivity. We anticipate that the use of this rapid, sensitive, and simple LAMP assay, which is the first of its kind for the identification and screening of C. jejuni in cattle farm samples, may play an important role in the prevention of C. jejuni contamination in the food chain, thereby reducing the risk of human campylobacteriosis.

Visual and Real-Time Event-Specific Loop-Mediated Isothermal Amplification Based Detection Assays for Bt Cotton Events MON531 and MON15985

2015 ◽  
Vol 98 (5) ◽  
pp. 1207-1214 ◽  
Author(s):  
Gurinder Jit Randhawa ◽  
Rashmi Chhabra ◽  
Rajesh K Bhoge ◽  
Monika Singh
Keyword(s):  
Real Time ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Bt Cotton ◽  
Loop Mediated Isothermal Amplification ◽  
Commercial Cultivation ◽  
Detection And Identification ◽  
Cotton Seeds ◽  
Gm Detection ◽  
Time Loop

Abstract Bt cotton events MON531 and MON15985 are authorized for commercial cultivation in more than 18 countries. In India, four Bt cotton events have been commercialized; more than 95% of total area under genetically modified (GM) cotton cultivation comprises events MON531 and MON15985. The present study reports on the development of efficient event-specific visual and real-time loop-mediated isothermal amplification (LAMP) assays for detection and identification of cotton events MON531 and MON15985. Efficiency of LAMP assays was compared with conventional and real-time PCR assays. Real-time LAMP assay was found time-efficient and most sensitive, detecting up to two target copies within 35 min. The developed real-time LAMP assays, when combined with efficient DNA extraction kit/protocol, may facilitate onsite GM detection to check authenticity of Bt cotton seeds.

scholarly journals Loop-Mediated Isothermal Amplification (LAMP) Assay to De-tect Toxoplasmosis in Schizophrenia Patients

2020 ◽  
Author(s):  
Hadi MIRAHMADI ◽  
Raheleh HASANZADEH ◽  
Hamid MALEK RAEESI ◽  
Shirzad FALLAHI ◽  
Mahdi KHOSHSIMA SHAHRAKI ◽  
...  
Keyword(s):  
Nested Pcr ◽  
High Efficiency ◽  
Parasitic Infection ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Lamp Method ◽  
Intermediate Hosts ◽  
Cross Sectional ◽  
Loop Mediated Isothermal Amplification ◽  
Schizophrenic Patients

Background: Toxoplasma gondii (T. gondii) causes an important parasitic infection known as toxoplasmosis, which is a globally distributed important zoonosis. One of the major serious characteristics of T. gondii is its ability to manipulate the behavior of intermediate hosts. We performed a cross-sectional study to determine toxoplasmosis in schizophrenic patients, as one of the major neuropsychiatric disorders, using loop-mediated isothermal amplification (LAMP) technic by targeting parasite B1 gene. Methods: Blood samples were taken from 118 schizophrenic patients hospitalized in tow hospitals including Baharan, Clinic of Psychiatric Ali-ibn-Abi-Talib Hospital (in Zahedan City), and Amir-al Momenin Psychiatric Hospital (in Zabol City), Sistan and Baluchestan Province, southeast Iran in 2016. They were analyzed using LAMP, and compared with the previous data of nested-PCR and serology. Results: Out of the 118 schizophrenic individuals, 56 patients (47.4%) were found to be infected with T. gondii. The diagnosis of toxoplasmosis was confirmed in 41 patients (34.7%) via the nested-PCR. The seroprevalence of toxoplasmosis in schizophrenic patients was 55.9% (66/118). Conclusion: We found a high efficiency of LAMP method in identifying toxoplasmosis and its high prevalence among schizophrenic patients. Our findings could provide viable offer implications for the prevention of schizophrenia.

Development and evaluation of a reverse transcription-loop-mediated isothermal amplification assay for rapid and high-sensitive detection of Cryptosporidium in water samples

2009 ◽  
Vol 60 (8) ◽  
pp. 2167-2172 ◽  
Author(s):  
A. Inomata ◽  
N. Kishida ◽  
T. Momoda ◽  
M. Akiba ◽  
S. Izumiyama ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Water Samples ◽  
Cryptosporidium Oocyst ◽  
Lamp Assay ◽  
Test Tube ◽  
Isothermal Amplification ◽  
Sensitive Detection ◽  
Loop Mediated Isothermal Amplification ◽  
Cryptosporidium Oocysts ◽  
Amplification Assay

We describe a novel assay for simple, rapid and high-sensitive detection of Cryptosporidium oocysts in water samples using a reverse transcription-loop-mediated isothermal amplification (RT-LAMP). The assay is based on the detection of 18S rRNA specific for Cryptosporidium oocysts. The detection limit of the developed RT-LAMP assay was as low as 6 × 10−3 oocysts/test tube, which theoretically enables us to detect a Cryptosporidium oocyst and perform duplicated tests even if water samples contain only one oocyst. The developed RT-LAMP assay could more sensitively detect Cryptosporidium oocysts in real water samples than the conventional assay based on microscopic observation.

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