Rapid Detection of SARS-CoV-2 Using Duplex Reverse Transcription-Multienzyme Isothermal Rapid Amplification in a Point-of-Care Testing

Chen, Hui; Sun, Chang; Wang, Yan; Gao, Xudong; You, Jinwei; Yu, Wanwan; Sun, Ningning; Yang, Yang; Li, Xiaojun

doi:10.3389/fcimb.2021.678703

Cited by 12 publications

(12 citation statements)

References 39 publications

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“…Agarose gel electrophoresis, real-time fluorescence, and lateral flow strip assay can be used to analyze MIRA results. Although several studies have used the MIRA technique to detect various infectious diseases, such as SARS-CoV-2 ( Chen et al, 2021 ), Mycobacterium tuberculosis ( Lu et al, 2021 ), and hepatitis B ( Sun et al, 2021 ), this method has not been used to detect S. aureus . Therefore, we hope to establish a rapid detection of S. aureus through this method.…”

Section: Introductionmentioning

confidence: 99%

Rapid detection of Staphylococcus aureus using a novel multienzyme isothermal rapid amplification technique

et al. 2022

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Staphylococcus aureus is a common pathogen that causes various infections. Therefore, it is crucial to develop a fast and easy detection method for diagnosing and preventing S. aureus infections. In this study, MIRA assay was developed and validated (specificity; 100%) for the detection of S. aureus with nuc as the target gene. The reaction temperature and reaction time were then optimized, and the best reaction was at 40°C, 20 min. The assay could detect S. aureus in only 25 min. Additionally, the limit of detection of MIRA was 5 × 102 CFU/ml, 10-fold lower than that of the traditional PCR. Furthermore, this assay efficiently detected 219 S. aureus of 335 strains obtained from different bacterial samples (detection accuracy; 99.40%). In conclusion, this study provides a rapid and easy-to-operate method for the detection of S. aureus, and thus can be used for the timely diagnosis and prevention of S. aureus infection.

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Section: Introductionmentioning

confidence: 99%

Rapid detection of Staphylococcus aureus using a novel multienzyme isothermal rapid amplification technique

et al. 2022

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“… Chen et al (2021a) designed a LFIA test strip based on a Duplex Reverse Transcription-Multienzyme Isothermal Rapid Amplification (RT-MIRA) for the visualization of the N and ORF1ab genes of SARS-CoV-2. The method eliminates the need for RNA extraction and requires only 2 steps for the detection of nasal or pharyngeal swab samples.…”

Section: Lifa Technique For Sars-cov-2 Nucleic Acid Detectionmentioning

confidence: 99%

Rapid detection of SARS-CoV-2: The gradual boom of lateral flow immunoassay

Zhu

Zhou

et al. 2023

Front. Bioeng. Biotechnol.

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Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is still in an epidemic situation, which poses a serious threat to the safety of people and property. Rapid diagnosis and isolation of infected individuals are one of the important methods to control virus transmission. Existing lateral flow immunoassay techniques have the advantages of rapid, sensitive, and easy operation, and some new options have emerged with the continuous development of nanotechnology. Such as lateral flow immunoassay test strips based on colorimetric-fluorescent dual-mode and gold nanoparticles, Surface Enhanced Raman Scattering, etc., these technologies have played an important role in the rapid diagnosis of COVID-19. In this paper, we summarize the current research progress of lateral flow immunoassay in the field of Severe Acute Respiratory Syndrome Coronavirus 2 infection diagnosis, analyze the performance of Severe Acute Respiratory Syndrome Coronavirus 2 lateral flow immunoassay products, review the advantages and limitations of different detection methods and markers, and then explore the competitive CRISPR-based nucleic acid chromatography detection method. This method combines the advantages of gene editing and lateral flow immunoassay and can achieve rapid and highly sensitive lateral flow immunoassay detection of target nucleic acids, which is expected to be the most representative method for community and clinical point-of-care testing. We hope that researchers will be inspired by this review and strive to solve the problems in the design of highly sensitive targets, the selection of detection methods, and the enhancement of CRISPR technology, to truly achieve rapid, sensitive, convenient, and specific detection of novel coronaviruses, thus promoting the development of novel coronavirus diagnosis and contributing our modest contribution to the world’s fight against epidemics.

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“…In this study, we present a novel diagnostic approach for T. vaginalis , combining MIRA, CRISPR/Cas13a, and lateral flow device (LFD) targeting. Isothermal amplification simplifies the nucleic acid amplification process by eliminating the need for thermal cycling, thus reducing the complexity and cost [ 19 ]. CRISPR/Cas13a is known for its high sensitivity and specificity in RNA targeting and cleavage, allowing for accurate identification of the pathogen [ 20 ].…”

Section: Introductionmentioning

confidence: 99%

A novel detection method based on MIRA-CRISPR/Cas13a-LFD targeting the repeated DNA sequence of Trichomonas vaginalis

Yang,

Wang,

et al. 2024

Parasites Vectors

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Background Trichomonas vaginalis is a protozoan parasite, widely recognized as the most prevalent non-viral sexually transmitted infection (STI) globally. This infection is linked to various complications, including pelvic inflammatory disease, adverse pregnancy outcomes, and an increased risk of acquiring HIV. Current molecular detection methods for T. vaginalis are often costly and technically challenging. Methods We developed a novel detection method for T. vaginalis using a multi-enzyme isothermal rapid amplification–clustered regularly interspaced short palindromic repeats (MIRA-CRISPR)/Cas13a-lateral flow device (LFD). This assay targets the repeated DNA sequence (GenBank: L23861.1) of T. vaginalis and is performed at a constant temperature of 37 °C for approximately 1 hour. Results The detection limit of genomic DNA (gDNA) using our protocol was 1 × 10–4 ng/μl. Specificity was confirmed by the absence of cross-reaction with gDNA from various other microorganisms such as Staphylococcus aureus, Lactobacillus taiwanensis, Escherichia coli, Monilia albicans, Giardia lamblia, or Toxoplasma gondii. Among 30 clinical samples tested, the positive rates of T. vaginalis detection were 33.33% (10/30) by wet mount microscopy, 40% (12/30) by nested polymerase chain reaction (PCR), 40% (12/30) by MIRA-CRISPR/Cas13a-LFD, and 40% (12/30) by the culture method. Compared with the culture method, the gold standard for diagnosing trichomoniasis, wet mount microscopy showed a sensitivity of 83.3% and moderate diagnostic agreement (kappa value = 0.87). Both nested PCR and MIRA-CRISPR/Cas13a-LFD exhibited 100% sensitivity and excellent diagnostic agreement (kappa value = 1). Conclusions The MIRA-CRISPR/Cas13a-LFD method is a convenient, rapid, stable, and accurate diagnostic tool for detecting T. vaginalis. This method has the potential to enhance the diagnosis and management of vaginitis, offering a significant improvement over existing diagnostic techniques. Graphical Abstract

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Rapid Detection of SARS-CoV-2 Using Duplex Reverse Transcription-Multienzyme Isothermal Rapid Amplification in a Point-of-Care Testing

Cited by 12 publications

References 39 publications

Rapid detection of Staphylococcus aureus using a novel multienzyme isothermal rapid amplification technique

Rapid detection of Staphylococcus aureus using a novel multienzyme isothermal rapid amplification technique

Rapid detection of SARS-CoV-2: The gradual boom of lateral flow immunoassay

A novel detection method based on MIRA-CRISPR/Cas13a-LFD targeting the repeated DNA sequence of Trichomonas vaginalis

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