A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of toxigenic Fusarium temperatum in maize stalks and kernels

Shan, Liuying; Haseeb, Hafiz Abdul; Zhang, Jun; Zhang, Dandan; Jeffers, Daniel; Dai, Xiaofeng; Guo, Wanshou

doi:10.1016/j.ijfoodmicro.2018.11.021

Cited by 17 publications

(4 citation statements)

References 22 publications

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“…The designed primer set LAMP could only amplify A. fabacearum , indicating its good specificity. By detecting the sensitivity of LAMP, we concluded that the LAMP primer set had a high sensitivity, and the lowest detection limit reached 5 × 10 −7 ng/μL This detection limit is lower than previously reported for the detection of potato late blight ( P. infestans ), blue mold decay ( Penicillium expansum ), maize ear and stalk rot diseases ( Fusarium temperatum ) and dried chickpea root rot ( Rhizoctonia bataticola ) [ 29 , 35 , 50 , 51 ]. The LAMP method established in this study is more precise and more sensitive than conventional PCR.…”

Section: Discussionmentioning

confidence: 81%

First Report of Crown Gall of Kiwifruit (Actinidia deliciosa) Caused by Agrobacterium fabacearum in China and the Establishment of Loop-Mediated Isothermal Amplification Technique

Shi

Zhang

et al. 2021

IJMS

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Kiwifruit is moderately sweet and sour and quite popular among consumers; it has been widely planted in some areas of the world. In 2019, the crown gall disease of kiwifruit was discovered in the main kiwifruit-producing area of Guizhou Province, China. This disease can weaken and eventually cause the death of the tree. The phylogeny, morphological and biological characteristics of the bacteria were described, and were related to diseases. The pathogenicity of this species follows the Koch hypothesis, confirming that A. fabacearum is the pathogen of crown gall disease of kiwifruit in China. In this study, Loop-mediated isothermal amplification (LAMP) analysis for genome-specific gene sequences was developed for the specific detection of A. fabacearum. The detection limit of the LAMP method is 5 × 10−7 ng/μL, which has high sensitivity. At the same time, the amplified product is stained with SYBR Green I after the reaction is completed, so that the amplification can be detected with the naked eye. LAMP analysis detected the presence of A. fabacearum in the roots and soil samples of the infected kiwifruit plant. The proposed LAMP detection technology in this study offers the advantages of ease of operation, visibility of results, rapidity, accuracy and high sensitivity, making it suitable for the early diagnosis of crown gall disease of kiwifruit.

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Section: Discussionmentioning

confidence: 81%

First Report of Crown Gall of Kiwifruit (Actinidia deliciosa) Caused by Agrobacterium fabacearum in China and the Establishment of Loop-Mediated Isothermal Amplification Technique

Shi

Zhang

et al. 2021

IJMS

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“…Fusarium species are important plant pathogens in many plants all over the world (Summerell et al, 2010;Srivastava et al, 2018;Shan et al, 2019). Fusarium spp.…”

Section: Resultsmentioning

confidence: 99%

Fusarium incarnatum Causing Fusarium Wilt on Protea (Protea cynaroides L.) in Turkey

Çat

2022

Türkiye Tarımsal Araştırmalar Dergisi

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Protea (Protea cynaroides L.) is one of the most important ornamental plants commercially cultivated in Turkey and other parts of the world. In May 2017, protea plants heavily showed wilting symptoms were obtained from the protea growing areas at Hatay province of Turkey. Brown vascular tissue in the stems of the infected plants was observed. To determine the causal agent of the disease, necrotic tissue pieces taken from diseased plants were surface-sterilized with 10% sodium hypochlorite, two times rinsed with sterile distilled water, placed on Petri dishes containing potato dextrose agar (PDA), and incubated at 24 oC for seven days. Microconidia were no septate, ovoid, hyaline, single-celled, and 9 to 12 × 1 to 3 μm. Macroconidia were slightly curved, four to six septate, and 27 to 31 × 3 to 5 μm. Based on the morphology and cultural features, the fungus was identified as F. incarnatum, which belongs to the F. incarnatum-equiseti species complex (FIESC). In addition, a pathogenicity test was made using the apple fruits. The pathogen was re-isolated from inoculated apple fruits and both cultural and morphological characteristics of the pathogen were identical. To confirm the identity of one representative isolate, the internal transcribed spacer (ITS) region including 5.8S rDNA was amplified and sequenced with primers of ITS-1 and ITS-4. Fusarium MLST, Fusarium-ID and GenBank database were used for the identification of sequence. The amplified 465 bp product has 99% nucleotide identity with the sequences of F. incarnatum-equiseti species complex of MLST types and deposited in GenBank under accession number (MH005097). According to the literature, this is the first molecular identification of F. incarnatum on protea plants in Turkey.

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“…within 60 min, as shown in Fig 4 . Based on the estimated molecular number of genomic DNA of M. mycetomatis, which is calculated to be 26.6 copies/pg, assuming a genome size of 36.7 Mbp [26]. The system detection limits are comparable not only to those targeting other fungi, such as Aspergillus [27] and Fusarium [28] but also to those of real-time PCR methods for Madurella (up to 3 pg) [16]. Furthermore, the sensitivity of our LAMP system is approximately 200 times higher than that reported previously [18].…”

Section: Discussionmentioning

confidence: 99%

“…It has been demonstrated that the LAMP reaction can be used with the crude fungal DNA extracted from clinical specimens [29][30][31], infected plants, or a few conidia with a simple treatment [28,32] due to its outstanding sensitivity and specificity. In addition, the preparation of crude DNA derived from mycetoma specimens for PCR diagnosis has been reported [13,16].…”

Section: Discussionmentioning

confidence: 99%

Specific and sensitive loop-mediated isothermal amplification (LAMP) method for Madurella strains, eumycetoma filamentous fungi causative agent

Yoshioka,

Mori,

Fahal

et al. 2023

PLoS Negl Trop Dis

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Background Filamentous fungi of the genus Madurella are the primary causative agents of mycetoma, a disease observed in tropical and subtropical regions. Since early diagnostics based on a morphological approach are difficult and have many shortcomings, a molecular diagnostic method suitable for rural settings is required. In this study, we developed the loop-mediated isothermal amplification (LAMP) method to present a foundational technique of the diagnosis of Madurella spp. (M. mycetomatis, M. pseudomycetomatis, M. tropicana, and M. fahalii), the common causative organisms of eumycetoma. Principal findings We successfully designed a primer pair targeting the rDNAs of three Madurella spp. excluding M. fahalii, and detected up to 100 fg of genomic DNA extracted from isolates of M. mycetomatis and 1 pg of M. pseudomycetomatis and M. tropicana, within one hour. Second, a primer pair specific to M. mycetomatis, the most common causative species, or M. fahalii, a drug-resistant species, was constructed, and the detection limit of both primer pairs was 1 pg. The designed primers accurately distinguished 16 strains of the genus Madurella from various fungal species known to cause mycetomas. Conclusion In summary, we established the first model of a LAMP detection method that rapidly and sensitively detects and identifies Madurella isolates for clinical diagnostics. Moreover, the combined designed primer sets could identify mycetoma-causing strains simultaneously.

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A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of toxigenic Fusarium temperatum in maize stalks and kernels

Cited by 17 publications

References 22 publications

First Report of Crown Gall of Kiwifruit (Actinidia deliciosa) Caused by Agrobacterium fabacearum in China and the Establishment of Loop-Mediated Isothermal Amplification Technique

First Report of Crown Gall of Kiwifruit (Actinidia deliciosa) Caused by Agrobacterium fabacearum in China and the Establishment of Loop-Mediated Isothermal Amplification Technique

Fusarium incarnatum Causing Fusarium Wilt on Protea (Protea cynaroides L.) in Turkey

Specific and sensitive loop-mediated isothermal amplification (LAMP) method for Madurella strains, eumycetoma filamentous fungi causative agent

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