Development of a nucleic acid lateral flow strip for rapid, visual detection of Nosema bombycis in silkworm eggs

He, Zhangshuai; Ni, Qi; Song, Yue; Wang, Rui; Tang, Yunlin; Wu, Yujiao; Liu, Lulu; Bao, Jialing; Chen, Jie; Long, Mengxian; Wei, Junhong; Li, Chunfeng; Li, Tian; Zhou, Zeyang; Pan, Guoqing

doi:10.1016/j.jip.2019.04.004

Cited by 14 publications

(7 citation statements)

References 23 publications

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“…NALFS's sensitivity for N. bombycis genomic DNA was 1 pg or 100 spores, which was 100 times more than the conventional PCR as reported by He et al. (2019).…”

Section: Immunodiagnostics‐based Detection Techniquesmentioning

confidence: 68%

“…The established method produced a good performance in detection as it was practical and effective. NALFS's sensitivity for N. bombycis genomic DNA was 1 pg or 100 spores, which was 100 times more than the conventional PCR as reported by He et al (2019).…”

Section: Nucleic Acid Lateral Flow Stripmentioning

confidence: 73%

“…A user‐friendly detection technique based on PCR and colloidal gold strip combined with a nucleic acid lateral flow strip (NALFS) was developed by He et al. (2019). The NALFS combines molecular and immunological technologies.…”

Section: Immunodiagnostics‐based Detection Techniquesmentioning

confidence: 99%

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Molecular diagnostics in sericulture: A paradigm shift towards disease diagnosis in silkworms

Deepika,

Ramesh,

Kumar

et al. 2024

Entomologia Exp Applicata

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Sericulture, the practice of rearing silkworms for the production of silk, is an essential agro‐based industry in several countries. However, silkworms are susceptible to a variety of diseases caused by viruses, bacteria and parasites, which may have a significant negative impact on global silk production. Traditional methods of pathogen identification, such as microscopy and laboratory culturing, have limitations in terms of accuracy and efficiency. The development of molecular techniques for pathogen identification has revolutionised the field of sericulture over the last decade. Genomic DNA and RNA‐based molecular techniques allow for the rapid and accurate detection of disease‐causing pathogens in silkworms. Molecular diagnosis has several advantages over traditional methods, including increased sensitivity and specificity, shorter turnaround time and the ability to detect pathogens that are difficult to culture or visualise under a microscope. Molecular techniques have been applied to detect several important pathogens of silkworms, including Nosema sp., nucleopolyhedrovirus, cypovirus, iflavirus and bidensovirus. The use of molecular diagnostics in sericulture is immensely important as the demand for high‐quality silk increases globally and the assessment of emerging pathogens associated with crop loss is essential. Major advancements in the improvement and application of molecular methods for diagnosing widespread silkworm pathogens are discussed.

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“…NALFS's sensitivity for N. bombycis genomic DNA was 1 pg or 100 spores, which was 100 times more than the conventional PCR as reported by He et al. (2019).…”

Section: Immunodiagnostics‐based Detection Techniquesmentioning

confidence: 68%

Section: Nucleic Acid Lateral Flow Stripmentioning

confidence: 73%

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Molecular diagnostics in sericulture: A paradigm shift towards disease diagnosis in silkworms

Deepika,

Ramesh,

Kumar

et al. 2024

Entomologia Exp Applicata

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“…As a result, many limitations associated with isothermal amplification are avoided, and since the multiplex-crRNA CRISPR/Cas12a detection system directly detects pathogens, nucleic acid amplification is unnecessary, enabling quantitative detection of pathogens. Our method could detect as low as 2 pg of pathogenic DNA, which would proximately equal to 2 × 10 2 spores in the system ( 32 ). This is a feasible and sensitive detection method.…”

Section: Discussionmentioning

confidence: 99%

Harnessing multiplex crRNA enables an amplification-free/CRISPR-Cas12a-based diagnostic methodology for Nosema bombycis

Zhang,

Zhao,

Cao

et al. 2024

Microbiol Spectr

Self Cite

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Nosema bombycis is transmitted horizontally and vertically from infected female moths to progeny eggs, bringing substantial economic losses to sericulture production. Timely and accurate detection of N. bombycis can effectively block the spread of silkworm pébrine disease, which is of great significance for its prevention and control. In this study, we established a multiplex-crRNA CRISPR/Cas12a nucleic acid amplification-free detection method for N. bombycis .. Specificity and sensitivity assessments confirmed that this method specifically detects N. bombycis without cross-reacting with other pathogens. It can detect N. bombycis at levels as low as 2 pg of genomic DNA. This method could also provide information on pathogen quantification, which will be important for in-field applications and promotions. IMPORTANCE The multiplex-crRNA CRISPR/Cas12a detection method saves hands-on time, reduces the risk of aerosol pollution, and can be directly applied to detecting silkworms infected with Nosema bombycis . This study provides a new approach for the inspection and quarantine of silkworm pébrine disease in sericulture and provides a new method for the detection of other pathogens.

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“…( 7) indicates the densities profile of a 2D Gaussian distribution so that its border resembles a 2D Gaussian distribution, in which the values of and are shown in Eq. (10).…”

Section: Nwd-lossmentioning

confidence: 99%

Intelligent Detection Method of Microparticle Virus in Silk-worm Based on YOLOv8 Improved Algorithm

Zhang,

Su,

Wang

et al. 2023

Preprint

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The presence of microparticle viruses seriously affects the quality of silkworm seed for domestic sericulture, so it is important to exclude them from detection in silkworm seed production. The traditional methods for detecting microparticle viruses in silkworms are manual microscopic observation, molecular biology, immunological approach, etc., which are cumbersome steps and cannot achieve intelligent detection and batch real-time detection. The YOLOv8 algorithm is used in this paper to address this problem. Firstly, NAM attention is introduced in the original algorithm's Backbone part. Therefore, the model can extract more generic information about features. Secondly, ODConv is used to replace Conv in the Head part of the original algorithm to enhance the model's ability to identify microparticle viruses. Finally, NWD-LOSS is used to modify the CIOU loss of the original algorithm to obtain a more accurate prediction box. The experiments indicate that the NN-YOLOv8 model is more accurate in detecting silkworm microparticle disease than mainstream detection algorithms. For a single image, the average detection time is 22.6 milliseconds, it has better prospects for future applications.

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Development of a nucleic acid lateral flow strip for rapid, visual detection of Nosema bombycis in silkworm eggs

Cited by 14 publications

References 23 publications

Molecular diagnostics in sericulture: A paradigm shift towards disease diagnosis in silkworms

Molecular diagnostics in sericulture: A paradigm shift towards disease diagnosis in silkworms

Harnessing multiplex crRNA enables an amplification-free/CRISPR-Cas12a-based diagnostic methodology for Nosema bombycis

Intelligent Detection Method of Microparticle Virus in Silk-worm Based on YOLOv8 Improved Algorithm

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