One-step detection of microRNA with high sensitivity and specificity via target-triggered loop-mediated isothermal amplification (TT-LAMP)

Sun, Yuanyuan; Tian, Hui; Liu, Chenghui; Sun, Yueying; Li, Zhengping

doi:10.1039/c7cc06140d

Cited by 74 publications

(28 citation statements)

References 27 publications

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“…Several other technologies have the potential to be alternative platforms suitable for small RNA detection. In particular, the LAMP assay, which has been previously developed for all three livestock trypanosome species based on gDNA targets [13, 40, 41] and for which a test intended for field application was developed for HAT [14], has been shown to be suitable for application to small RNA as an assay substrate [42, 43] and therefore could potentially be optimised to develop a field-applicable 7SL-sRNA assay. Another recently developed method, Recombinase Polymerase Amplification (e.g.…”

Section: Discussionmentioning

confidence: 99%

Parasite specific 7SL-derived small RNA is an effective target for diagnosis of active trypanosomiasis infection

et al. 2019

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Human and animal African trypanosomiasis (HAT & AAT, respectively) remain a significant health and economic issue across much of sub-Saharan Africa. Effective control of AAT and potential eradication of HAT requires affordable, sensitive and specific diagnostic tests that can be used in the field. Small RNAs in the blood or serum are attractive disease biomarkers due to their stability, accessibility and available technologies for detection. Using RNAseq, we have identified a trypanosome specific small RNA to be present at high levels in the serum of infected cattle. The small RNA is derived from the non-coding 7SL RNA of the peptide signal recognition particle and is detected in the serum of infected cattle at significantly higher levels than in the parasite, suggesting active processing and secretion. We show effective detection of the small RNA in the serum of infected cattle using a custom RT-qPCR assay. Strikingly, the RNA can be detected before microscopy detection of parasitaemia in the blood, and it can also be detected during remission periods of infection when no parasitaemia is detectable by microscopy. However, RNA levels drop following treatment with trypanocides, demonstrating accurate prediction of active infection. While the small RNA sequence is conserved between different species of trypanosome, nucleotide differences within the sequence allow generation of highly specific assays that can distinguish between infections with Trypanosoma brucei, Trypanosoma congolense and Trypanosoma vivax. Finally, we demonstrate effective detection of the small RNA directly from serum, without the need for pre-processing, with a single step RT-qPCR assay. Our findings identify a species-specific trypanosome small RNA that can be detected at high levels in the serum of cattle with active parasite infections. This provides the basis for the development of a cheap, non-invasive and highly effective diagnostic test for trypanosomiasis.

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Section: Discussionmentioning

confidence: 99%

Parasite specific 7SL-derived small RNA is an effective target for diagnosis of active trypanosomiasis infection

et al. 2019

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“…Herein, Sun et al. [96] put forward a new idea that simplifies the probe design. In this work, a stem-loop template DNA and a stem-loop primer were designed instead of the original overlong template.…”

Section: Newly Developed Methodsmentioning

confidence: 99%

Research advances in the detection of miRNA

Tian

et al. 2019

Journal of Pharmaceutical Analysis

289

152

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MicroRNAs (miRNAs) are a family of endogenous, small (approximately 22 nucleotides in length), noncoding, functional RNAs. With the development of molecular biology, the research of miRNA biological function has attracted significant interest, as abnormal miRNA expression is identified to contribute to serious human diseases such as cancers. Traditional methods for miRNA detection do not meet current demands. In particular, nanomaterial-based methods, nucleic acid amplification-based methods such as rolling circle amplification (RCA), loop-mediated isothermal amplification (LAMP), strand-displacement amplification (SDA) and some enzyme-free amplifications have been employed widely for the highly sensitive detection of miRNA. MiRNA functional research and clinical diagnostics have been accelerated by these new techniques. Herein, we summarize and discuss the recent progress in the development of miRNA detection methods and new applications. This review will provide guidelines for the development of follow-up miRNA detection methods with high sensitivity and specificity, and applicability to disease diagnosis and therapy.

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“…In the presence of miRNA and DNA polymerase, a strand displacement DNA synthesis and exponential amplification can proceed. This method uses 4-6 primers and 6-8 target sequences [114]. EXPAR, combined with quantum-dot-based sensors, could be a sensitive method for miRNA amplification, even at low concentrations.…”

Section: Methods That Could Be Used In Transcriptome Studiesmentioning

confidence: 99%

Transcriptomics in Alzheimer’s Disease: Aspects and Challenges

Bagyinszky

Giau

2020

IJMS

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Alzheimer’s disease (AD) is the most common cause of dementia. Although the heritability of AD is high, the knowledge of the disease-associated genes, their expression, and their disease-related pathways remain limited. Hence, finding the association between gene dysfunctions and pathological mechanisms, such as neuronal transports, APP processing, calcium homeostasis, and impairment in mitochondria, should be crucial. Emerging studies have revealed that changes in gene expression and gene regulation may have a strong impact on neurodegeneration. The mRNA–transcription factor interactions, non-coding RNAs, alternative splicing, or copy number variants could also play a role in disease onset. These facts suggest that understanding the impact of transcriptomes in AD may improve the disease diagnosis and also the therapies. In this review, we highlight recent transcriptome investigations in multifactorial AD, with emphasis on the insights emerging at their interface.

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One-step detection of microRNA with high sensitivity and specificity via target-triggered loop-mediated isothermal amplification (TT-LAMP)

Abstract: A novel one-step microRNA assay is developed based on a target-triggered loop-mediated isothermal amplification (TT-LAMP) mechanism, which enables the accurate detection of as low as 100 aM (1 zmol) microRNA with simple one-step operation by using only one-type of DNA polymerase.

Cited by 74 publications

References 27 publications

Parasite specific 7SL-derived small RNA is an effective target for diagnosis of active trypanosomiasis infection

Parasite specific 7SL-derived small RNA is an effective target for diagnosis of active trypanosomiasis infection

Research advances in the detection of miRNA

Transcriptomics in Alzheimer’s Disease: Aspects and Challenges

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