amplification assay
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2022 ◽  
Vol 12 ◽  
Author(s):  
Yushan Bu ◽  
Wenjun Qiao ◽  
Zhengyuan Zhai ◽  
Tongjie Liu ◽  
Pimin Gong ◽  
...  

Raw milk is susceptible to microbial contamination during transportation and storage. Pseudomonas fluorescens producing heat-resistant enzymes have become the most common and harmful psychrophilic microorganisms in the cold chain logistics of raw milk. To rapidly detect P. fluorescens in raw milk, the protease gene aprX was selected as a detection target to construct a set of primers with strong specificity, and a loop-mediated isothermal amplification (LAMP) assay was established. The detection thresholds of the LAMP assay for pure cultured P. fluorescens and pasteurized milk were 2.57 × 102 and 3 × 102 CFU/mL, respectively. It had the advantages over conventional method of low detection threshold, strong specificity, rapid detection, and simple operation. This LAMP assay can be used for online monitoring and on-site detection of P. fluorescens in raw milk to guarantee the quality and safety of dairy products.


Author(s):  
Gamil S. G. Zeedan ◽  
Abeer M. Abdalhamed ◽  
Raafat M. Shaapan ◽  
Amira H. El-Namaky

Abstract Background This study was conducted to detect the presence of T. gondii in milk and blood samples using three different assays: enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and loop-mediated isothermal amplification assay (LAMP). Whole blood, serum, and milk samples were collected from goats (n = 156), sheep (n = 261), and camels (n = 108) in different governorates in Egypt from December 2019 to February 2021 and screened by ELISA for anti-Toxoplasma IgG antibodies before DNA extraction. The target T. gondii DNA gene was detected and evaluated using the LAMP assay compared to PCR. Results T. gondii antibodies were found in milk and serum samples at the rates of (29.26%) and (36.58%) in camels, (34.18%) and (35.89%) in sheep, and (33.7%) and (36.36%) in goats, respectively. Similar to PCR, the percentages of LAMP tests for the detection of the T. gondii DNA gene in milk and blood samples of camels, sheep, and goats were (4.8, 14.63), (6.83, 7.69), and (7.79, 9.09), respectively. LAMP's sensitivity for detecting T. gondii in milk and blood samples, which was identical to that of PCR, was 100%. Conclusions The findings clearly demonstrated that there were no variations in T. gondii detection capabilities in milk and blood samples from various animals using both PCR and LAMP tests. It provides a quick, precise, and sensitive method of detecting T. gondii in a variety of samples that may be used both in the field and in laboratory diagnosis.


2022 ◽  
Vol 14 (11) ◽  
Author(s):  
Dong Chen ◽  
Fangju Tian ◽  
Wanyu Liu ◽  
Jingyi Yu ◽  
Dafeng Song

Background: Salmonella is one of the main foodborne bacterial pathogens, causing diseases and death. The study used reverse transcription loop-mediated isothermal amplification (RT-LAMP) to detect Salmonella. Objectives: To design six primers and detect Salmonella using RT-LAMP to facilitate the rapid detection of pathogenic bacteria in food. Methods: We designed six primers based on the gene coding sequences of inv A, specific to Salmonella. Each reaction solution contained 6.0 mM MgSO4, 1 M betaine, 1.6 mM dNTPs, 160 U/mL Bst DNA polymerase, 0.2 μM of both external primers, 0.8 μM of both internal primers, and 0.2 μM of both loop primers. The reaction temperature was 65°C. Results: Our amplified products were separated by 2% agarose gel electrophoresis. The detection limit was 10 CFU per reaction. Conclusions: RT-LAMP exhibited the same accuracy as the GB assay in detecting Salmonella in foods. RT-LAMP was highly specific and sensitive; hence, it may serve as an effective tool in detecting Salmonella.


Author(s):  
Zhi-shan Feng ◽  
Jing-yi Li ◽  
Jing-yun Zhang ◽  
Feng-yu Li ◽  
Hong-xia Guan ◽  
...  

2021 ◽  
Vol 7 (12) ◽  
pp. 115333-115354
Author(s):  
Sarah Amado Ribeiro ◽  
Calebe Bertolino Marins De Campos ◽  
Hérida Samaya Gonçalves De Sousa ◽  
Alex Silva Da Cruz ◽  
Aparecido Divino Da Cruz

The objective of the study was to carry out, with the aid of Scopus®️, a scientometric analysis of Loop Mediated Isothermal Amplification Assay (LAMP) applied to farm animals. The research has considered articles from January 2000 to December 2019 and only open or closed access articles published in English. The bibliometric matrices were run through RStudio, applying Biblioshiny as a web interface to Bibliometrix resources for R environment. Later, several bibliometric data were collected with the aid of Bibliometrix, most of which were converted into graphs using Microsoft Excel®️. The scientometric analysis base of the current study was composed by 438 articles from 504 researched in the Scopus®️. Of the 438 articles analyzed, it stands out as results: 1) the years of 2015 (11,4%) and 2019 (11,4%) had equally the highest number of publications in the area; 2) Journal of Virological Methods (12,5%) ranked first in the ranking of journals according to total articles published; 3) China (49,8%), Japan (12,7%) and India (7,1%) have been countries of more published articles; 4) most articles applied the assay to detect microorganisms affecting the farm animals; and, 5) together, the animal groups fish, bovine, poultry, and swine corresponded to 2/3 (71,1%) of the animals used in scientific research using the LAMP method. With all these results, it is concluded that the scientometric analysis showed an overview of the information in the articles about LAMP applied to farm animals.


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