2020
DOI: 10.1016/j.jmoldx.2020.08.003
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Multiplex High-Resolution Melting Assay for Simultaneous Identification of Molecular Markers Associated with Extended-Spectrum Cephalosporins and Azithromycin Resistance in Neisseria gonorrhoeae

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Cited by 21 publications
(17 citation statements)
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“…Currently, the comprehensive approach for genetic mutation monitoring is next‐generation sequencing (NGS); however, it is costly, time‐consuming and required expertise of bioinformatics (Ji et al ., 2020 ). PCR coupled with Sanger sequencing has also been previously described as a method for detecting mutations (Xiu et al ., 2020 ). However, due to the large number of mutation sites in SARS‐CoV‐2, analysing these sites by singleplex PCR requires very repetitive manual pipetting, therefore increase the risk of cross‐contamination.…”
Section: Discussionmentioning
confidence: 99%
See 1 more Smart Citation
“…Currently, the comprehensive approach for genetic mutation monitoring is next‐generation sequencing (NGS); however, it is costly, time‐consuming and required expertise of bioinformatics (Ji et al ., 2020 ). PCR coupled with Sanger sequencing has also been previously described as a method for detecting mutations (Xiu et al ., 2020 ). However, due to the large number of mutation sites in SARS‐CoV‐2, analysing these sites by singleplex PCR requires very repetitive manual pipetting, therefore increase the risk of cross‐contamination.…”
Section: Discussionmentioning
confidence: 99%
“…In this study, we propose a methodology based on high‐resolution melting (HRM) analysis for SARS‐CoV‐2 detection that can rapidly and consistently identify the virus, subtypes and mutations without requiring a sequencing analysis step, thereby avoiding the need for reagents and/or supplies. HRM is a convenient, extensible, reliable, closed‐tube and cost‐effective approach that has been used for identification of species or mutation scanning or molecular typing in several research fields, such as epidemiology and microbiology (Montgomery et al ., 2010 ; Tong and Giffard, 2012 ; Li et al ., 2014 ; Xiu et al ., 2020b ; Xiu et al ., 2020 ). By exploring the advantages of HRM analysis, we developed a multiplex assay using reverse transcription coupled with HRM analysis of the amplification products to rapidly detect viral RNA and simultaneously identify mutations in SARS‐CoV‐2.…”
Section: Introductionmentioning
confidence: 99%
“…1) [18]. For N. gonorrhoeae detection, we selected porA as the target as was frequently used to identity N. gonorrhoeae in other methods [17,[38][39][40]. Based on the characteristics of its single-base resolution, we constructed two crRNA sequences that could identify A2059G and C2611T separately.…”
Section: Schematic Of Cas13a Based N Gonorrhoeae Detection and Azithromycin Resistance Identificationmentioning
confidence: 99%
“…This novel method is based on a high-resolution melting (HRM) analysis with unlabeled probes and complements current M. genitalium detection using RT-PCR and WGS. HRM is a convenient, closed-tube, and cost-efficient method that is widely used in several research fields, including variant scanning, species identification, and molecular typing ( 19 22 ). Although HRM is superior to RT-PCR for identifying many small insertions or deletions and complex mutations, differentiating between two or more possible single nucleotide polymorphisms at a site can be problematic when probes are not used ( 23 , 24 ).…”
Section: Introductionmentioning
confidence: 99%