scholarly journals Detection of Potato ring rot Pathogen Clavibacter michiganensis subsp. sepedonicus by Loop-mediated isothermal amplification (LAMP) assay

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Hasan Sagcan ◽  
Neslihan Turgut Kara
Keyword(s):  
Rapid Diagnosis ◽  
Isothermal Amplification ◽  
Diagnostic Methods ◽  
Economic Losses ◽  
Temperature Cycle ◽  
Lamp Method ◽  
Clavibacter Michiganensis ◽  
Loop Mediated Isothermal Amplification ◽  
Ring Rot ◽  
Potato Ring Rot

AbstractClavibacter michiganensis subsp. sepedonicus (CMS) is an important bacterial plant pathogen causing potato ring rot disease. Rapid diagnosis of CMS is crucial because of the economic losses caused by serious harvest losses. Although there are serological tests used in the rapid diagnosis of CMS, they are not widely used because of their low sensitivity. The DNA-based PCR methods, which are highly sensitive, do not have the possibility of on-site diagnosis, especially since they require serious laboratory infrastructure. In recent years, scientists have been working on alternative amplification methods to develop DNA-based point of care (POC) diagnostic methods. Accordingly, the loop-mediated isothermal amplification (LAMP) method, which was developed in the early 2000s, provides an important convenience for DNA-based tests to use in the field. Due to the unique design of primers, more amplification products could be create in a shorter time than conventional amplification methods without needing a temperature cycle, and it can be applied with the aid of a simple heater without requiring a laboratory environment. In this study, efficient LAMP method for the detection of CMS has optimized. For device-independent detection of LAMP products, colorimetric method and LFD has used.

scholarly journals Detection of Haemophilus parasuis isolates from South China by loop-mediated isothermal amplification and isolate characterisation

2012 ◽  
Vol 79 (1) ◽  
Author(s):  
Jian-min Zhang ◽  
Hai-yan Shen ◽  
Ming Liao ◽  
Tao Ren ◽  
Li-li Guo ◽  
...  
Keyword(s):  
16S Rrna ◽  
South China ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Economic Losses ◽  
Lamp Method ◽  
Haemophilus Parasuis ◽  
Loop Mediated Isothermal Amplification ◽  
Glässer’S Disease ◽  
Glasser's Disease

Haemophilus parasuis is the etiological agent of Glässer’s disease, which is characterised by fibrinous polyserositis, meningitis and polyarthritis, causing severe economic losses to the swine industry. In this study, a loop-mediated isothermal amplification (LAMP) test was developed to improve the specificity, facility and speed of diagnosis of H. parasuis isolates. The LAMP assay rapidly amplified the target gene within 50 min incubation at 63 °C in a laboratory water bath. The LAMP amplicon could be visualised directly in the reaction tubes following the addition of SYBR Green I dye. The detection limit of this LAMP method was 10 CFU/mL, which was 10 times more sensitive than the earlier 16S rRNA polymerase chain reaction (PCR) test conducted by Oliveira, Galina and Pijoan (2001), and no cross-reactivity was observed from other non-H. parasuis strains. This LAMP test was evaluated further on 187 clinical specimens from pigs suspected of being infected with H. parasuis. Forty-three were found positive by bacterial isolation of H. parasuis, as well as by the 16S rRNA PCR and LAMP tests. The 43 H. parasuis isolates were classified into 9 serovars and had 37 genetic patterns when analysed by pulsed-field gel electrophoresis (PFGE). This displayed that various H. parasuis serovars and genotypes were widely distributed in South China. Therefore, the speed, specificity and sensitivity of the LAMP test, the lack of a need for expensive equipment, and the visual readout showed great potential for a correct clinical diagnosis of H. parasuis in favour of controlling Glässer’s disease.

scholarly journals Development of a loop-mediated isothermal amplification (LAMP) method for specific detection of Mycobacterium bovis

2021 ◽  
Vol 15 (1) ◽  
pp. e0008996
Author(s):  
Thoko Flav Kapalamula ◽  
Jeewan Thapa ◽  
Mwangala Lonah Akapelwa ◽  
Kyoko Hayashida ◽  
Stephen V. Gordon ◽  
...  
Keyword(s):  
Developing Countries ◽  
Mycobacterium Bovis ◽  
Color Change ◽  
Low Cost ◽  
Lamp Assay ◽  
Genomic Region ◽  
Isothermal Amplification ◽  
Diagnostic Methods ◽  
Lamp Method ◽  
Loop Mediated Isothermal Amplification

Bovine tuberculosis (TB) caused by Mycobacterium bovis is a significant health threat to cattle and a zoonotic threat for humans in many developing countries. Rapid and accurate detection of M. bovis is fundamental for controlling the disease in animals and humans, and for the proper treatment of patients as one of the first-line anti-TB drug, pyrazinamide, is ineffective against M. bovis. Currently, there are no rapid, simplified and low-cost diagnostic methods that can be easily integrated for use in many developing countries. Here, we report the development of a loop-mediated isothermal amplification (LAMP) assay for specific identification of M. bovis by targeting the region of difference 4 (RD4), a 12.7 kb genomic region that is deleted solely in M. bovis. The assay's specificity was evaluated using 139 isolates comprising 65 M. bovis isolates, 40 M. tuberculosis isolates, seven M. tuberculosis complex reference strains, 22 non-tuberculous mycobacteria and five other bacteria. The established LAMP detected only M. bovis isolates as positive and no false positives were observed using the other mycobacteria and non-mycobacteria tested. Our LAMP assay detected as low as 10 copies of M. bovis genomic DNA within 40 minutes. The procedure of LAMP is simple with an incubation at a constant temperature. Results are observed with the naked eye by a color change, and there is no need for expensive equipment. The established LAMP can be used for the detection of M. bovis infections in cattle and humans in resource-limited areas.

scholarly journals Loop-mediated isothermal amplification assay forEnterococcus sp., E. coliandS. aureusin chicken

2018 ◽  
Author(s):  
HyeSoon Song ◽  
YouChan Bae ◽  
HyukMan Kwon ◽  
YongKuk Kwon ◽  
SeongJoon Joh
Keyword(s):  
Broiler Chicken ◽  
Simultaneous Detection ◽  
Isothermal Amplification ◽  
Diagnostic Methods ◽  
Economic Losses ◽  
Diagnostic Assays ◽  
E Coli ◽  
Loop Mediated Isothermal Amplification ◽  
Resource Limited ◽  
Amplification Assay

ABSTRACTBacterial chondronecrosis with osteomyelitis (BCO) is a major cause of lameness in broiler chicken, and results in serious economic losses worldwide. Although the pathogenesis mechanism leading to lameness is not entirely understood, some strains ofEnterococcus sp., avian pathogenicEscherichia coli, orStaphylococcus aureushave been long recognized as important causative pathogens. To prevent the progression ofEnterococcus sp., avian pathogenicE. coli, orS. aureusinfections, we developed rapid, sensitive, and convenient diagnostic assays using loop-mediated isothermal amplification (LAMP). Entero-Common-LAMP assays were developed for a simultaneous detection of eightEnterococcusspecies. To target specific microorganisms, seven Entero-Specific-LAMP assays forE. faecalis, E. faecium, E. hirae, E. gallinarum, E. avium, E. duransandE. cecorum, andE. coli-LAMP andS. aureus-LAMP assays, were developed. Considering the prevalence and economic impact ofEnterococcus sp., E. coli, andS. aureus, the developed ten different LAMP assays have a considerable potential as routine diagnostic methods in the field or in resource-limited environments.

scholarly journals Fast, Precise, and Reliable Multiplex Detection of Potato Viruses by Loop-Mediated Isothermal Amplification

2020 ◽  
Vol 21 (22) ◽  
pp. 8741
Author(s):  
Güven Edgü ◽  
Lena Julie Freund ◽  
Stefanie Hartje ◽  
Eckhard Tacke ◽  
Hans-Reinhard Hofferbert ◽  
...  
Keyword(s):  
Disease Management ◽  
Management Practices ◽  
Tobacco Rattle Virus ◽  
Simultaneous Detection ◽  
Isothermal Amplification ◽  
Diagnostic Methods ◽  
Tuber Quality ◽  
Loop Mediated Isothermal Amplification ◽  
Tuber Necrosis ◽  
Corky Ringspot

Potato is an important staple food crop in both developed and developing countries. However, potato plants are susceptible to several economically important viruses that reduce yields by up to 50% and affect tuber quality. One of the major threats is corky ringspot, which is a tuber necrosis caused by tobacco rattle virus (TRV). The appearance of corky ringspot symptoms on tubers prior to commercialization results in ≈ 45% of the tubers being downgraded in quality and value, while ≈ 55% are declared unsaleable. To improve current disease management practices, we have developed simple diagnostic methods for the reliable detection of TRV without RNA purification, involving minimalized sample handling (mini), subsequent improved colorimetric loop-mediated isothermal amplification (LAMP), and final verification by lateral-flow dipstick (LFD) analysis. Having optimized the mini-LAMP-LFD approach for the sensitive and specific detection of TRV, we confirmed the reliability and robustness of this approach by the simultaneous detection of TRV and other harmful viruses in duplex LAMP reactions. Therefore, our new approach offers breeders, producers, and farmers an inexpensive and efficient new platform for disease management in potato breeding and cultivation.

Rapid and sensitive diagnosis of adenoviral keratoconjunctivitis by loop-mediated isothermal amplification (LAMP) method

2004 ◽  
Vol 28 (6) ◽  
pp. 445-450 ◽  
Author(s):  
Taketoshi Wakabayashi ◽  
Ryoko Yamashita ◽  
Tetsuhiko Kakita ◽  
Mito Kakita ◽  
Tetsuro Oshika
Keyword(s):  
Isothermal Amplification ◽  
Lamp Method ◽  
Loop Mediated Isothermal Amplification
Sign in / Sign up

Export Citation Format

Share Document