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.

scholarly journals Feasibility and Performance of Loop-Mediated Isothermal Amplification Assay in the Diagnosis of Pulmonary Tuberculosis in Decentralized Settings in Eastern China

2019 ◽  
Vol 2019 ◽  
pp. 1-4
Author(s):  
Zhongdong Wang ◽  
Haiyan Sun ◽  
Zhisheng Ren ◽  
Bai Xue ◽  
Jie Lu ◽  
...  
Keyword(s):  
Eastern China ◽  
Isothermal Amplification ◽  
Smear Microscopy ◽  
Loop Mediated Isothermal Amplification ◽  
Resource Limited ◽  
Control And Prevention ◽  
Amplification Assay ◽  
Smear Negative ◽  
And Performance ◽  
Culture Positive

Early diagnosis is essential for the control and prevention of tuberculosis (TB). The objective of this study was to investigate the feasibility and performance of loop-mediated isothermal amplification (LAMP) in the diagnosis of pulmonary TB in county-level microscopy centers in Qingdao, Eastern China. A total of 523 presumptive TB patients were consecutively recruited between July 2017 and April 2018, and 22 patients were excluded from the analysis. Of 102 culture-positive cases, TB-LAMP identified 91 cases, demonstrating a sensitivity of 89.2%. In comparison, the sensitivity of routine smear microscopy was 69.6% (71/102), which was significantly lower than that of TB-LAMP (P=0.001). In addition, TB-LAMP sensitivities in smear-positive and smear-negative samples were 98.6% and 67.7%, respectively. In conclusion, our data demonstrate that TB-LAMP outperforms conventional smear microscopy in TB diagnosis, which could be used as an alternative method for smear microscopy in resource-limited settings in China.

scholarly journals Enhancing colorimetric loop-mediated isothermal amplification speed and sensitivity with guanidine chloride

BioTechniques ◽  
2020 ◽  
Vol 69 (3) ◽  
pp. 178-185 ◽  
Author(s):  
Yinhua Zhang ◽  
Guoping Ren ◽  
Jackson Buss ◽  
Andrew J Barry ◽  
Gregory C Patton ◽  
...  
Keyword(s):  
Guanidine Hydrochloride ◽  
Isothermal Amplification ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Simple Method ◽  
Diagnostic Assays ◽  
Loop Mediated Isothermal Amplification ◽  
Dna And Rna ◽  
Versatile Technique ◽  
Primer Sets

Loop-mediated isothermal amplification (LAMP) is a versatile technique for detection of target DNA and RNA, enabling rapid molecular diagnostic assays with minimal equipment. The global SARS-CoV-2 pandemic has presented an urgent need for new and better diagnostic methods, with colorimetric LAMP utilized in numerous studies for SARS-CoV-2 detection. However, the sensitivity of colorimetric LAMP in early reports has been below that of the standard RT-qPCR tests, and we sought to improve performance. Here we report the use of guanidine hydrochloride and combined primer sets to increase speed and sensitivity in colorimetric LAMP, bringing this simple method up to the standards of sophisticated techniques and enabling accurate, high-throughput diagnostics.

scholarly journals Earlyin plantadetection ofXanthomonas axonopodispv.punicaein pomegranate using enhanced loop-mediated isothermal amplification assay

2018 ◽  
Author(s):  
M.K. PrasannaKumar ◽  
P. Buela Parivallal ◽  
C. Manjunath ◽  
H.B. Mahesh ◽  
Karthik S Narayan ◽  
...  
Keyword(s):  
Post Infection ◽  
Bacterial Blight ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Economic Losses ◽  
Specific Primers ◽  
Loop Mediated Isothermal Amplification ◽  
Negative Results ◽  
Hydroxynaphthol Blue ◽  
Amplification Assay

AbstractBacterial blight in pomegranate caused byXanthomonas axonopodispv.punicae(Xap) is an increasing threat for pomegranate cultivation in India. To prevent the economic losses, it is pivotal to detect the infection in latent stages rather than in later stages. We have developed an enhanced method termed as loop-mediated isothermal amplification (LAMP) technique to evaluate for the latent detection of Xap in pomegranate using six set of specific primers. Three DNA intercalating dyes were used, such as Ethidium bromide, hydroxynaphthol blue (HNB) and SYBR Green resulted in visualising the positivity for LAMP assay. The reaction time and temperature were to be 65°C from 30 min onwards, for the dyes and its sensitivity was observed up to 10−7ng in the LAMP assay. For field applicability, LAMP assay detected Xap on 7thday post infection while the PCR amplified Xap after 11thday post infection. Finally, the specificity of LAMP assay was validated to be positive with ten Xap isolates for its accuracy and 29 non-Xap bacterial isolates showed negative results. Moreover, this method could be used as a better alternative to PCR based methods, for early detection of the pathogens.

scholarly journals Loop-Mediated Isothermal Amplification Assay for Detection of Generic and Verocytotoxin-ProducingEscherichia coliamong Indigenous Individuals in Malaysia

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Cindy Shuan Ju Teh ◽  
Kek Heng Chua ◽  
Yvonne Ai Lian Lim ◽  
Soo Ching Lee ◽  
Kwai Lin Thong
Keyword(s):  
Large Scale ◽  
Lamp Assay ◽  
Asymptomatic Infection ◽  
Isothermal Amplification ◽  
Indigenous Populations ◽  
Bacterial Strains ◽  
E Coli ◽  
Loop Mediated Isothermal Amplification ◽  
Threshold Time ◽  
Amplification Assay

We have successfully developed a Loop-mediated isothermal amplification(LAMP) assay that could specifically detect genericEscherichia coli(E. coli). This assay was tested on 85 bacterial strains and successfully identified 54E. colistrains (average threshold time, Tt = 21.26). The sensitivity of this assay was evaluated on serial dilutions of bacterial cultures and spiked faeces. The assay could detect 102 CFU/mL for bacterial culture with Tt = 33.30 while the detection limit for spiked faeces was 103 CFU/mL (Tt = 31.12). We have also detected 46 genericE. colifrom 50 faecal samples obtained from indigenous individuals with 16% of the positive samples being verocytotoxin-producingE. coli(VTEC) positive. VT1/VT2 allele was present in one faecal sample while the ratio of VT1 to VT2 was 6 : 1. Overall, our study had demonstrated high risk of VTEC infection among the indigenous community and most of the asymptomatic infection occurred among those aged below 15 years. The role of asymptomatic human carriers as a source of dissemination should not be underestimated. Large scale screening of the VTEC infection among indigenous populations and the potential contamination sources will be possible and easy with the aid of this newly developed rapid and simple LAMP assay.

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