Rapid detection of human enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) is important in the early phase of hand-foot-and-mouth disease (HFMD). In this study, we developed and evaluated a novel reverse transcription–isothermal multiple-self-matching-initiated amplification (RT-IMSA) assay for the rapid detection of EV71 and CVA16 by use of reverse transcriptase, together with a strand displacement DNA polymerase. Real-time RT-IMSA assays using a turbidimeter and visual RT-IMSA assays to detect EV71 and CVA16 were established and completed in 1 h, and the reported corresponding real-time reverse transcription–loop-mediated isothermal amplification (RT-LAMP) assays targeting the same regions of the VP1 gene were adopted as parallel tests. Through testing VP1 RNAs transcribed in vitro, the real-time RT-IMSA assays exhibited better linearity of quantification, with R2 values of 0.952 (for EV71) and 0.967 (for CVA16), than the real-time RT-LAMP assays, which had R2 values of 0.803 (for EV71) and 0.904 (for CVA16). Additionally, the detection limits of the real-time RT-IMSA assays (approximately 937 for EV71 and 67 for CVA16 copies/reaction) were higher than those of real-time RT-LAMP assays (approximately 3,266 for EV71 and 430 for CVA16 copies/reaction), and similar results were observed in the visual RT-IMSA assays. The new approaches also possess high specificities for the corresponding targets, with no cross-reactivity observed. In clinical assessment, compared to commercial reverse transcription-quantitative PCR (qRT-PCR) kits, the diagnostic sensitivities of the real-time RT-IMSA assays (96.4% for EV71 and 94.6% for CVA16) were higher than those of the real-time RT-LAMP assays (91.1% for EV71 and 90.8% for CVA16). The visual RT-IMSA assays also exhibited the same results. In conclusion, this proof-of-concept study suggests that the novel RT-IMSA assay is superior to the RT-LAMP assay in terms of detection limit and has the potential to rapidly detect EV71 and CVA16 viruses.

Improved Detection Limit in Rapid Detection of Human Enterovirus 71 and Coxsackievirus A16 by a Novel Reverse Transcription–Isothermal Multiple-Self-Matching-Initiated Amplification Assay

Evaluation of a rapid detection for Coxsackievirus B3 using one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP)

Foot-and-mouth disease (FMD) is a viral disease. FMD diagnosis in the field is based on clinical signs that are shared by other vesicular diseases, hence to confirm FMD a laboratory is needed. Laboratory diagnostic techniques including serology may fail to distinguish between vaccinated and new infection, virus isolation may take up to 4 days to yield results, while molecular techniques including PCR, which are accurate, sensitive, specific and rapid, are costly and require special training of the laboratory staff. These challenges limit laboratory diagnosis yet in Uganda FMD outbreaks are common since the disease is endemic. This work reports the comparative detection of Foot-and-mouth disease virus (FMDV) by reverse transcription-loop mediated isothermal amplification (RT-LAMP) and Real-Time polymerase chain reaction (rRT-PCR) in Uganda based on the 3D polymerase (3D pol ) gene. The rRT-PCR assay is considered as the gold standard. A total of 89 cattle samples that included epithelial tissues (16.9%) and oral swabs (83.1%) were collected from outbreak cases in Eastern Districts of Mbale and Budaka. These were applied to molecular assays of rRT-PCR and RT- LAMP using primers and probes targeting the 3D pol gene. The diagnostic sensitivity and specificity of RT-LAMP as a screening test and rRT-PCR as the reference test was 94.44% (95% CI = 94.11 to 94.78%) and 98.59% (95% CI = 98.50 to 98.68%), respectively. The kappa value for diagnostic agreement between rRT-PCR and RT-LAMP was 93.0% (95% CI = 83.50 to 100%), showing a perfect agreement. In conclusion, the RT-LAMP assay had a very high sensitivity and specificity when compared to the reference test of rRT-PCR. It was also very rapid since it gave results in 45 to 60 min. Due to its simplicity, sensitivity and specificity, LAMP assay has the potential for use in routine surveillance of FMD in Uganda.

/pdf/comparative-detection-of-foot-and-mouth-disease-virus-by-37m5wzp15k.pdf

Comparative detection of foot-and-mouth disease virus by reverse transcription loop-mediated isothermal amplification assay and real time polymerase chain reaction in Uganda

The invention discloses a real-time isothermal amplification-based kit for human papilloma virus E6/E7 gene detection, and application thereof. A primer and a probe are designed by aiming at 14 high-risk HPV, and rapid detection and identification on HPV E6/E7 mRNA can be realized specifically. The kit provided by the invention has the characteristics of high speed, high efficiency, sensitivity, specificity, real-time detection and analysis and the like, provides a rapid and accurate molecular detection method for generation investigation of HPV and prevention and treatment of cervical cancer, greatly reduces the inspection cost and has an important popularization and application value.

Real-time isothermal amplification-based kit for human papilloma virus E6/E7 gene detection, and application thereof

Typing and quantitative detection method and kit for human papilloma virus (HPV)

A sensitive reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for human enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) infection was further evaluated The one step reaction was performed in a single tube at 65?C for 45 min for EV71 and 35 min for CVA16 The detection limits of RT-LAMP assays for both EV71 and CVA16 were 01 of a 50% tissue culture infective dose (TCID50) per reaction, based on 10—Fold dilutions of a titrated EV71 or CVA16 strain The specific assay showed there were no cross-reactions with Coxsackievirus A (CVA) viruses (CVA 2, 4, 5, 7, 9, 10, 14, and 25), Coxsackievirus B (CVB) viruses (CVB 1, 2, 3, 4, and 5) or ECHO viruses (ECHO 3, 6, 11, and 19) In parallel with commercial quantitative real-time polymerase chain reaction (qRT-PCR) diagnostic kits for EV71 and CVA16, the RT-LAMP assay was evaluated with 515 clinical specimens, the results showed the RT-LAMP assay and the qRT-PCR assay were in complete agreement for 513/515 (996%) of the specimens Two samples with discrepant results from two methods were further verified by nested reverse transcription polymerase chain reaction (nRT-PCR) assay and sequencing to be true positives for CVA16 In conclusion, RT-LAMP assay is demonstrated to be a sensitive and specific assay and have a great potential for the rapid and visual screening of EV71 and CVA16 in China, especially in those resource-limited hospitals and rural clinics of provincial and municipal regions

/pdf/evaluation-of-reverse-transcription-loop-mediated-isothermal-2mn1diww6r.pdf

Evaluation of Reverse Transcription Loop-Mediated Isothermal Amplification assays for Rapid Detection of Human Enterovirus 71 and Coxsackievirus A16 in Clinical Samples

The invention belongs to the biotechnological application field, and relates to the detection and the monitoring of Chinese-common six HPVs of cervical scraping smear specimens of suspected HPV (human papilloma virus) infected patients with genital damage (comprising genital warts and cervical epithelial neoplasia) as main symptoms in disease prevention control mechanisms at all levels, papilloma virus detection network laboratories, sentinel point hospitals and the like. The conserved sequences of E6, E7 or L1 genes of six HPV subtypes are analyzed by bioinformatics professional software through applying gene color based loop-mediated isothermal amplification (LAMP) technology, six primers of each subtype, which completely couple with eight binding domains in each target sequence, are designed, single reverse transcription and nested PCR secondary application steps are omitted, an HNB (hydroxynaphthol blue) dye is added before the reaction, the amplification in a 63DEG C isothermal environment for 1h can be completed only through one step, and a detection result can be determined through the color change observed with naked eyes and the white precipitate at the bottom of a tube. Compared with traditional PCR methods, the technology provided in the invention has the advantages of safety, specificity, sensitivity, and convenience.

Color based loop-mediated isothermal amplification technology for detecting HPV6, HPV11, HPV16, HPV18, HPV52 and HPV58

The invention which belongs to the biotechnological application field relates to a research on GeXP multi-PCR technology for detecting and parting HPV, and concretely relates to the detection and the typing of markers of cervical secretions and the like infected with various HPV subtypes (comprising HPV6, HPV11, HPV31, HPV33 and HPV52) in disease prevention control mechanisms at all levels, hospitals and the like. The research is concretely carried out by analyzing conserved sequences of HPV subtype genes with computer software, respectively designing specific primers aiming at the subtypes, and carrying out single-tube multi-PCR (six-PCR) (comprising Rnasep-DNA), wherein the whole reaction lasts for less than 2h. The research which overcomes a disadvantage of no parting of routine single-tube multiple fluorescent qualitative PCR detections and also overcomes disadvantages of complex operation, long time, high cost and the like of routine chip detection methods provides a new idea for HPV parting technologies, and characteristics of high specificity, high sensitivity and rapidness of the research provide strong technological supports for the realization of the rapid and accurate screening and discrimination of HPV subtype infection, so the research has important meanings to research the HPV infection type distribution characteristic of a patient and improve the control level of cancer pathologies caused by the HPV subtypes.

Research on GeXP multi-PCR technology for detecting and parting human papilloma virus (HPV)

Handle everyday research tasks with reliable, citation-backed results

Your personal Research Agent to handle research tasks with citation-backed results

Popular Tasks used by Researchers

How can I help with your research?

Meet SciSpace

Get more enhanced response by uploading the PDFs you want me to reference.

No relevant PDFs in your library

SciSpace is the AI research assistant for academics. Run systematic literature reviews on 280M+ papers, and write papers with cited sources. Trusted by 1M+ students, PhDs & researchers.

SciSpace | AI for Research

Analyze PDFs

Code & Manuscripts

Funding & Grants

Literature & Patents

Medical & Clinical Data

Systematic Review

Visualize & Present

Web & Data

Build a Google Scholar-like website for your research.

Build a website

Create charts and images for your research

Create a Chart

Write a paper for submission to a journal

Draft a manuscript

Patent Search

Design eye-catching scientific posters in minutes.

Scientific Poster Generation

Systematic Literature Review

One task is running at the moment. Your messages will be shown right after.

Drag and drop or click here to browse

Loved by <highlight>1 million+</highlight> researchers

Extract a list of specific topics and their sources from unstructured text

Topics

Compare and analyze relevant papers that matches with your search

Papers

Get insights from PDFs and bookmarked papers from your library

My library

Recent searches

Try searching for:

Catch AI-generated content in scholarly and non-scholarly content

{ai} Detector

Ai Writer

Get PDF Summaries, highlighted text explanations 

Chat with PDF

Effortlessly create in-text citations and bibliographies in APA and 2,500 other formats

Citation generator

Get explanations, summaries, and answers on academic papers

Ease up your research workflow with {scispace}'s cohort of exciting AI tools

Elevate your academic writing skills and convey your ideas the way you want

Paraphraser

Explore our range of reading and writing tools

Your file is being prepared and should be ready in a few minutes. If it's a large file, it might take a bit longer. You can close this window, and we'll email you the file when it's done.

You have reached a maximum limit of <strong>{limit}</strong> columns in the table. Remove at least <strong>1</strong> column to add or create another one.

Mengjie Yang

Author Tools

Chat about Author

Papers

Evaluation of Reverse Transcription Loop-Mediated Isothermal Amplification assays for Rapid Detection of Human Enterovirus 71 and Coxsackievirus A16 in Clinical Samples

Color based loop-mediated isothermal amplification technology for detecting HPV6, HPV11, HPV16, HPV18, HPV52 and HPV58

Research on GeXP multi-PCR technology for detecting and parting human papilloma virus (HPV)