High-performance detection of Mycobacterium bovis in milk using digital LAMP

Tao, Yi; Yun, Juanli; Wang, Jian; Xu, Peng; Li, Caiming; Liu, Hongtao; Lan, Ying; Pan, Jian-Zhang; Du, Wei

doi:10.1016/j.foodchem.2020.126945

Cited by 27 publications

(9 citation statements)

References 28 publications

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“…A previous study reported that Mycobacterium was also the dominant pathogens from the water supply network and typical urban surface water (Wolf-Baca and Piekarska 2020), which was positively related to autochthonous organisms (Leight et al 2018). Several species of Mycobacterium were pathogenic to both animals and human beings, causing pulmonary and cutaneous disease, like tuberculosis (Jin et al 2018;Tao et al 2020). In contrast, Liang et al (2020) found the higher abundance of Burkholderia-Paraburkholderia, Bacillales and Staphylococcus in the bioaerosols samples at the same area.…”

Section: Association With Human Activitymentioning

confidence: 92%

Pollution profile of waterborne bacterial and fungal community in urban Rivers of Pearl River estuary: Microbial safety assessment

Wang

et al. 2021

Journal of Freshwater Ecology

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Waterborne pathogens in urban rivers of estuarine areas pose a great threat to human health. In this study, both bacterial and fungal community structures of four urban rivers in the Pearl River estuarine area were investigated using high-throughput sequencing. Results showed that the diversity and richness indices of the bacterial community were higher than that of fungi. Taxonomic analysis indicated that the most abundant bacterial phylum was Actinobacteria, while the most abundant fungal phylum was Ascomycota. In addition, 35 major pathogens were found at the level of genus with Mycobacterium being the most abundant. Specifically, pathogens of Burkholderiales, Bartonella, Rhizobiales, Bordetella and Brucella were first detected in this area, although with relative low abundances. Spearman correlation analysis revealed that temperature, TOC and TDP were significant factors shaping both total microbial community and pathogenic microbial community structure. TDP was positively correlated with almost all the potential pathogenic genera. PICRUSt analysis further predicted that bacterial infectious disease was the most abundant pathway associated with microbial functions of human disease in all the studied rivers. This work provides useful information not only for in-depth understanding of bacterial and fungal communities in urban rivers of estuarine areas under anthropogenic disturbances but also for environmental risk management of potential waterborne pathogens.

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Section: Association With Human Activitymentioning

confidence: 92%

Pollution profile of waterborne bacterial and fungal community in urban Rivers of Pearl River estuary: Microbial safety assessment

Wang

et al. 2021

Journal of Freshwater Ecology

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“…Among molecular diagnostic methods, LAMP has emerged as a promising test and has been successfully applied in the diagnosis of mycobacterium diseases, including M. tuberculosis, M. bovis, M. kansasii and M. avium (Thapa, Maharjan et al 2019;Yashiki, Yamazaki et al 2019;Chen, Lu et al 2020;Das and Panigrahi 2020;Tao, Yun et al 2020;Das and Panigrahi, 2020). The LAMP technique used in this study is highly specific; it can differentiate M. leprae from M. tuberculosis and 8 other non-tuberculosis Mycobacteria.…”

Section: Discussionmentioning

confidence: 99%

Loop-mediated isothermal amplification (LAMP) assay targeting RLEP for detection of Mycobacterium leprae in leprosy patients

Jiang

Tsang

Wang

et al. 2021

International Journal of Infectious Diseases

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Leprosy is a chronic infectious disease caused by Mycobacterium leprae and it remains a significant health problem in several parts of the world. Early and accurate diagnosis of this disease is therefore essential. Previously published loop-mediated isothermal amplification (LAMP) protocols for detecting mycobacterial species used conventional primers targeting the 16S rRNA, gyrB and insertion sequence genes. Methods: In this study, we conducted a LAMP assay for leprosy and compared it with quantitative polymerase chain reaction (q-PCR) and conventional PCR assays to determine the efficiency, sensitivity and specificity of each technique. We chose conserved sequence RLEP as a suitable molecular target for assays. Results: The LAMP assay provided rapid and accurate results, confirming leprosy in 91/110 clinical skin tissue samples from leprosy patients and amplifying the target pathogen in <60 min at 65 C. The assay was more sensitive than conventional PCR and more straightforward and faster than the q-PCR assay. Conclusions: The LAMP assay has the potential for developing quicker, more accessible visual methods for the detection of M. leprae, which will enable early diagnosis and treatment and prevent further infection in endemic areas.

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“…LAMP is conducted in the presence of Bst DNA polymerase, deoxyribonucleotide triphosphate (dNTP), and 4-6 specific primers (inner and outer primers) that recognize 6-8 specific regions [23]. The Bst DNA polymerase removes the need for high thermocycling because of its activity at 50-70 • C, and the DNA polymerase is less sensitive to food-origin inhibitors [24,25]. The use of two outer primers (forward outer primer; F3, and backward outer primer; B3), two inner primers (forward inner primer; FIP, and backward inner primer; BIP), with additional loop primers (forward loop primer; LF, and backward loop primer; LB) also allows sequence-specific detection, improves specificity, and accelerates assay specificity to amplification targets [26].…”

Section: Isothermal Amplification Methodsmentioning

confidence: 99%

“…However, complex food matrices can inhibit the detection of these food-borne pathogens. Therfore, food-origin inhibitors possess inhibitors that make the DNA/RNA extraction challenging, denature DNA or polymerases, or bind magnesium ions to interfere with amplification [22,24,25]. Furthermore, it has been reported that these inhibitors are complex polysaccharides, phenolic compounds, and calcium ions [19,26].…”

Section: Inhibitors That Originate From Food During the Isothermal Amplification Processmentioning

confidence: 99%

A Review of Isothermal Amplification Methods and Food-Origin Inhibitors against Detecting Food-Borne Pathogens

Moon

Lee

2022

Foods

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The isothermal amplification method, a molecular-based diagnostic technology, such as loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA), is widely used as an alternative to the time-consuming and labor-intensive culture-based detection method. However, food matrices or other compounds can inhibit molecular-based diagnostic technologies, causing reduced detection efficiencies, and false-negative results. These inhibitors originating from food are polysaccharides and polyphenolic compounds in berries, seafood, and vegetables. Additionally, magnesium ions needed for amplification reactions can also inhibit molecular-based diagnostics. The successful removal of inhibitors originating from food and molecular amplification reaction is therefore proposed to enhance the efficiency of molecular-based diagnostics and allow accurate detection of food-borne pathogens. Among molecular-based diagnostics, PCR inhibitors have been reported. Nevertheless, reports on the mechanism and removal of isothermal amplification method inhibitors are insufficient. Therefore, this review describes inhibitors originating from food and some compounds inhibiting the detection of food-borne pathogens during isothermal amplification.

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High-performance detection of Mycobacterium bovis in milk using digital LAMP

Cited by 27 publications

References 28 publications

Pollution profile of waterborne bacterial and fungal community in urban Rivers of Pearl River estuary: Microbial safety assessment

Pollution profile of waterborne bacterial and fungal community in urban Rivers of Pearl River estuary: Microbial safety assessment

Loop-mediated isothermal amplification (LAMP) assay targeting RLEP for detection of Mycobacterium leprae in leprosy patients

A Review of Isothermal Amplification Methods and Food-Origin Inhibitors against Detecting Food-Borne Pathogens

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